790 resultados para Perceived control
Resumo:
Annual reports are an important component of New Zealand schools’ public accountability. Through the annual report the governance body informs stakeholders about school aims, objectives, achievements, use of resources, and financial performance. We examine the perceived usefulness of the annual report in the discharge of public accountability. We find that 15% of the recipients (mainly parents/caregivers) do not read the annual report because they trust in the school to do the right thing or rely on others to monitor school activities. We find that the annual report is used for a variety of purposes including to determine if the school has conducted its activities effectively and achieved stated objectives and goals; to examine student achievements; to assess financial accountability and performance; and to make decisions about the school as a suitable environment for their child/children. We find that other forms of communication such as school newsletters, parent-teacher interviews, children and other parents are more important sources of information about the school than the annual report.
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This study explored the psychological influences of hands-free and hand-held mobile phone use while driving. Participants were 796 Australian drivers aged 17 to 76 years who owned mobile phones. A cross-sectional survey assessed frequency of calling and text messaging while driving (overall, hands-free, hand-held) as well as drivers’ behavioural, normative, and control beliefs relating to mobile phone use while driving. Irrespective of handset type, 43% of drivers reported answering calls while driving on a daily basis, followed by making calls (36%), reading text messages (27%), and sending text messages (18%). In total, 63.9% of drivers did not own hands-free kits and, of the drivers that owned hand-free kits, 32% did not use it most or all of the time. Significant differences were found in the behavioural, normative, and control beliefs of frequent and infrequent users of both types of handset while driving. As expected, frequent users reported more advantages of, more approval from others for, and fewer barriers that would prevent them from, using either a hands-free or a hand-held mobile phone while driving than infrequent users. Campaigns to reduce mobile phone use while driving should attempt to minimise the perceived benefits of the behaviour and highlight the risks of this unsafe driving practice.
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Background: Exercise could contribute to weight loss by altering the sensitivity of the appetite regulatory system. Objective: The aim of this study was to assess the effects of 12 wk of mandatory exercise on appetite control. Design: Fifty-eight overweight and obese men and women [mean (±SD) body mass index (in kg/m2) = 31.8 ± 4.5, age = 39.6 ± 9.8 y, and maximal oxygen intake = 29.1 ± 5.7 mL · kg–1 · min–1] completed 12 wk of supervised exercise in the laboratory. The exercise sessions were designed to expend 2500 kcal/wk. Subjective appetite sensations and the satiating efficiency of a fixed breakfast were compared at baseline (week 0) and at week 12. An Electronic Appetite Rating System was used to measure subjective appetite sensations immediately before and after the fixed breakfast in the immediate postprandial period and across the whole day. The satiety quotient of the breakfast was determined by calculating the change in appetite scores relative to the breakfast's energy content. Results: Despite large variability, there was a significant reduction in mean body weight (3.2 ± 3.6 kg), fat mass (3.2 ± 2.2 kg), and waist circumference (5.0 ± 3.2 cm) after 12 wk. The analysis showed that a reduction in body weight and body composition was accompanied by an increase in fasting hunger and in average hunger across the day (P < 0.0001). Paradoxically, the immediate and delayed satiety quotient of the breakfast also increased significantly (P < 0.05). Conclusions: These data show that the effect of exercise on appetite regulation involves at least 2 processes: an increase in the overall (orexigenic) drive to eat and a concomitant increase in the satiating efficiency of a fixed meal.
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This research investigated underlying issues that were critical to the success of the bifocal trial and comprised of three studies. The first study evaluated if Chinese-Canadian children were suitable subjects for the bifocal trial. The high prevalence of myopia in Chinese children suggests that genetic input plays a role in myopia development, but the rapid increase in prevalence over the last few decades indicates environmental factors are also important. Since this bifocal trial was conducted in Canada, this work aimed to determine whether Chinese children who had migrated to Canada would still have high myopia prevalence and a high rate of myopia progression. The second study determined the optimal bifocal lens power for myopia treatment and the effect of incorporating base-in prism into the bifocal. In the majority of published myopia control studies, the power of the prescribed near addition was usually predetermined in the belief that the near addition would always help to improve the near focus. In fact, the effect of near addition on the accommodative error might be quite different even for individuals in which the same magnitude of accommodation lag had been measured. Therefore, this work was necessary to guide the selection of bifocal and prism powers most suitable for the subsequent bifocal trial. The third study, the ultimate goal of this research, was to conduct a longitudinal clinical trial to determine if bifocals and prismatic bifocals could control myopia progression in children.
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Abstract: Purpose – Several major infrastructure projects in the Hong Kong Special Administrative Region (HKSAR) have been delivered by the build-operate-transfer (BOT) model since the 1960s. Although the benefits of using BOT have been reported abundantly in the contemporary literature, some BOT projects were less successful than the others. This paper aims to find out why this is so and to explore whether BOT is the best financing model to procure major infrastructure projects. Design/methodology/approach – The benefits of BOT will first be reviewed. Some completed BOT projects in Hong Kong will be examined to ascertain how far the perceived benefits of BOT have been materialized in these projects. A highly profiled project, the Hong Kong-Zhuhai-Macau Bridge, which has long been promoted by the governments of the People's Republic of China, Macau Special Administrative Region and the HKSAR that BOT is the preferred financing model, but suddenly reverted back to the traditional financing model to be funded primarily by the three governments with public money instead, will be studied to explore the true value of the BOT financial model. Findings – Six main reasons for this radical change are derived from the analysis: shorter take-off time for the project; difference in legal systems causing difficulties in drafting BOT agreements; more government control on tolls; private sector uninterested due to unattractive economic package; avoid allegation of collusion between business and the governments; and a comfortable financial reserve possessed by the host governments. Originality/value – The findings from this paper are believed to provide a better understanding to the real benefits of BOT and the governments' main decision criteria in delivering major infrastructure projects.
Resumo:
This thesis by publication contributes to our knowledge of psychological factors underlying a modern day phenomenon, young people’s mobile phone behaviour. Specifically, the thesis reports a PhD program of research which adopted a social psychological approach to explore mobile phone behaviour among young Australians aged between 15 and 24 years. A particular focus of the research program was to explore both the cognitive and behavioural aspects of young people’s mobile phone behaviour which for the purposes of this thesis is defined as mobile phone involvement. The research program comprised three separate stages which were developmental in nature, in that, the findings of each stage of the research program informed the next. The overarching goal of the program of research was to improve our understanding of the psychosocial factors influencing young people’s mobile phone behaviour. To achieve this overall goal, there were a number of aims to the research program which reflect the developmental nature of this thesis. Given the limited research into the mobile phone behaviour in Australia, the first two aims of the research program were to explore patterns of mobile phone behaviour among Australian youth and explore the social psychological factors relating to their mobile phone behaviour. Following this exploration, the research program sought to develop a measure which captures the cognitive and behavioural aspects of mobile phone behaviour. Finally, the research program aimed to examine and differentiate the psychosocial predictors of young people’s frequency of mobile phone use and their level of involvement with their mobile phone. Both qualitative and quantitative methodologies were used throughout the program of research. Five papers prepared during the three stages of the research program form the bulk of this thesis. The first stage of the research program was a qualitative investigation of young people’s mobile phone behaviour. Thirty-two young Australians participated in a series of focus groups in which they discussed their mobile phone behaviour. Thematic data analysis explored patterns of mobile phone behaviour among young people, developed an understanding of psychological factors influencing their use of mobile phones, and identified that symptoms of addiction were emerging in young people’s mobile phone behaviour. Two papers (Papers 1 and 2) emanated from this first stage of the research program. Paper 1 explored patterns of mobile phone behaviour and revealed that mobile phones were perceived as being highly beneficial to young people’s lives, with the ability to remain in constant contact with others being particularly valued. The paper also identified that symptoms of behavioural addiction including withdrawal, cognitive and behavioural salience, and loss of control, emerged in participants’ descriptions of their mobile phone behaviour. Paper 2 explored how young people’s need to belong and their social identity (two constructs previously unexplored in the context of mobile phone behaviour) related to their mobile phone behaviour. It was revealed that young people use their mobile phones to facilitate social attachments. Additionally, friends and peers influenced young people’s mobile phone behaviour; for example, their choice of mobile phone carrier and their most frequent type of mobile phone use. These papers laid the foundation for the further investigation of addictive patterns of behaviour and the role of social psychological factors on young people’s mobile behaviour throughout the research program. Stage 2 of the research program focussed on developing a new parsimonious measure of mobile phone behaviour, the Mobile Phone Involvement Questionnaire (MPIQ), which captured the cognitive and behavioural aspects of mobile phone use. Additionally, the stage included a preliminary exploration of factors influencing young people’s mobile phone behaviour. Participants (N = 946) completed a questionnaire which included a pool of items assessing symptoms of behavioural addiction, the uses and gratifications relating to mobile phone use, and self-identity and validation from others in the context of mobile phone behaviour. Two papers (Papers 3 & 4) emanated from the second stage of the research program. Paper 3 provided an important link between the qualitative and quantitative components of the research program. Qualitative data from Stage 1 indicated the reasons young people use their mobile phones and identified addictive characteristics present in young people’s mobile phone behaviour. Results of the quantitative study conducted in Stage 2 of the research program revealed the uses and gratifications relating to young people’s mobile phone behaviour and the effect of these gratifications on young people’s frequency of mobile phone use and three indicators of addiction, withdrawal, salience, and loss of control. Three major uses and gratifications: self (such as feeling good or as a fashion item), social (such as contacting friends), and security (such as use in an emergency) were found to underlie much of young people’s mobile phone behaviour. Self and social gratifications predicted young people’s frequency of mobile phone use and the three indicators of addiction but security gratifications did not. These results provided an important foundation for the inclusion of more specific psychosocial predictors in the later stages of the research program. Paper 4 reported the development of the mobile phone involvement questionnaire and a preliminary exploration of the effect of self-identity and validation from others on young people’s mobile phone behaviour. The MPIQ assessed a unitary construct and was a reliable measure amongst this cohort. Results found that self-identity influenced the frequency of young people’s use whereas self-identity and validation from others influenced their level of mobile phone involvement. These findings provided an important indication that, in addition to self factors, other people have a strong influence on young people’s involvement with their mobile phone and that mobile phone involvement is conceptually different to frequency of mobile phone use. Stage 3 of the research program empirically examined the psychosocial predictors of young people’s mobile behaviour and one paper, Paper 5, emanated from this stage. Young people (N = 292) from throughout Australia completed an online survey assessing the role of self-identity, ingroup norm, the need to belong, and self-esteem on their frequency of mobile phone use and their mobile phone involvement. Self-identity was the only psychosocial predictor of young people’s frequency of mobile phone use. In contrast, self-identity, ingroup norm, and need to belong all influenced young people’s level of involvement with their mobile phone. Additionally, the effect of self-esteem on young people’s mobile phone involvement was mediated by their need to belong. These results indicate that young people who perceive their mobile phone to be an integral part of their self-identity, who perceive that mobile phone is common amongst friends and peers, and who have a strong need for attachment to others, in some cases driven by a desire to enhance their self-esteem, are most likely to become highly involved with their mobile phones. Overall, this PhD program of research has provided an important contribution to our understanding of young Australians’ mobile phone behaviour. Results of the program have broadened our knowledge of factors influencing mobile phone behaviour beyond the approaches used in previous research. The use of various social psychological theories combined with a behavioural addiction framework provided a novel examination of young people’s mobile behaviour. In particular, the development of a new measure of mobile phone behaviour in the research program facilitated the differentiation of the psychosocial factors influencing frequency of young people’s mobile phone behaviour and their level of involvement with their mobile phone. Results of the research program indicate the important role that mobile phone behaviour plays in young people’s social development and also signals the characteristics of those people who may become highly involved with their mobile phone. Future research could build on this thesis by exploring whether mobile phones are affecting traditional social psychological processes and whether the results in this research program are generalisable to other cohorts and other communication technologies.
Resumo:
Overweight and obesity are two of the most important emerging public health issues in our time and regarded by the World Health Organisation [WHO] (1998) as a worldwide epidemic. The prevalence of obesity in the USA is the highest in the world, and Australian obesity rates fall into second place. Currently, about 60% of Australian adults are overweight (BMI „d 25kg/m2). The socio-demographic factors associated with overweight and/or obesity have been well demonstrated, but many of the existing studies only examined these relationships at one point of time, and did not examine whether significant relationships changed over time. Furthermore, only limited previous research has examined the issue of the relationship between perception of weight status and actual weight status, as well as factors that may impact on people¡¦s perception of their body weight status. Aims: The aims of the proposed research are to analyse the discrepancy between perceptions of weight status and actual weight status in Australian adults; to examine if there are trends in perceptions of weight status in adults between 1995 to 2004/5; and to propose a range of health promotion strategies and furth er research that may be useful in managing physical activity, healthy diet, and weight reduction. Hypotheses: Four alternate hypotheses are examined by the research: (1) there are associations between independent variables (e.g. socio -demographic factors, physical activity and dietary habits) and overweight and/or obesity; (2) there are associations between the same independent variables and the perception of overweight; (3) there are associations between the same independent variables and the discrepancy between weight status and perception of weight status; and (4) there are trends in overweight and/or obesity, perception of overweight, and the discrepancy in Australian adults from 1995 to 2004/5. Conceptual Framework and Methods: A conceptual framework is developed that shows the associations identified among socio -demographic factors, physical activity and dietary habits with actual weight status, as well as examining perception of weight status. The three latest National Health Survey data bases (1995 , 2001 and 2004/5) were used as the primary data sources. A total of 74,114 Australian adults aged 20 years and over were recruited from these databases. Descriptive statistics, bivariate analyses (One -Way ANOVA tests, unpaired t-tests and Pearson chi-square tests), and multinomial logistic regression modelling were used to analyse the data. Findings: This research reveals that gender, main language spoken at home, occupation status, household structure, private health insurance status, and exercise are related to the discrepancy between actual weight status and perception of weight status, but only gender and exercise are related to the discrepancy across the three time point s. The current research provides more knowledge about perception of weight status independently. Factors which affect perception of overweight are gender, age, language spoken at home, private health insurance status, and diet ary habits. The study also finds that many factors that impact overweight and/or obesity also have an effect on perception of overweight, such as age, language spoken at home, household structure, and exercise. However, some factors (i.e. private health insurance status and milk consumption) only impact on perception of overweight. Furthermore, factors that are rel ated to people’s overweight are not totally related to people’s underestimation of their body weight status in the study results. Thus, there are unknown factors which can affect people’s underestimation of their body weight status. Conclusions: Health promotion and education activities should provide education about population health education and promotion and education for particular at risk sub -groups. Further research should take the form of a longitudinal study design ed to examine the causal relationship between overweight and/or obesity and underestimation of body weight status, it should also place more attention on the relationships between overweight and/or obesity and dietary habits, with a more comprehensive representation of SES. Moreover, further research that deals with identification of characteristics about perception of weight status, in particular the underestimation of body weight status should be undertaken.
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The study reported here, constitutes a full review of the major geological events that have influenced the morphological development of the southeast Queensland region. Most importantly, it provides evidence that the region’s physiography continues to be geologically ‘active’ and although earthquakes are presently few and of low magnitude, many past events and tectonic regimes continue to be strongly influential over drainage, morphology and topography. Southeast Queensland is typified by highland terrain of metasedimentary and igneous rocks that are parallel and close to younger, lowland coastal terrain. The region is currently situated in a passive margin tectonic setting that is now under compressive stress, although in the past, the region was subject to alternating extensional and compressive regimes. As part of the investigation, the effects of many past geological events upon landscape morphology have been assessed at multiple scales using features such as the location and orientation of drainage channels, topography, faults, fractures, scarps, cleavage, volcanic centres and deposits, and recent earthquake activity. A number of hypotheses for local geological evolution are proposed and discussed. This study has also utilised a geographic information system (GIS) approach that successfully amalgamates the various types and scales of datasets used. A new method of stream ordination has been developed and is used to compare the orientation of channels of similar orders with rock fabric, in a topologically controlled approach that other ordering systems are unable to achieve. Stream pattern analysis has been performed and the results provide evidence that many drainage systems in southeast Queensland are controlled by known geological structures and by past geological events. The results conclude that drainage at a fine scale is controlled by cleavage, joints and faults, and at a broader scale, large river valleys, such as those of the Brisbane River and North Pine River, closely follow the location of faults. These rivers appear to have become entrenched by differential weathering along these planes of weakness. Significantly, stream pattern analysis has also identified some ‘anomalous’ drainage that suggests the orientations of these watercourses are geologically controlled, but by unknown causes. To the north of Brisbane, a ‘coastal drainage divide’ has been recognized and is described here. The divide crosses several lithological units of different age, continues parallel to the coast and prevents drainage from the highlands flowing directly to the coast for its entire length. Diversion of low order streams away from the divide may be evidence that a more recent process may be the driving force. Although there is no conclusive evidence for this at present, it is postulated that the divide may have been generated by uplift or doming associated with mid-Cenozoic volcanism or a blind thrust at depth. Also north of Brisbane, on the D’Aguilar Range, an elevated valley (the ‘Kilcoy Gap’) has been identified that may have once drained towards the coast and now displays reversed drainage that may have resulted from uplift along the coastal drainage divide and of the D’Aguilar blocks. An assessment of the distribution and intensity of recent earthquakes in the region indicates that activity may be associated with ancient faults. However, recent movement on these faults during these events would have been unlikely, given that earthquakes in the region are characteristically of low magnitude. There is, however, evidence that compressive stress is building and being released periodically and ancient faults may be a likely place for this stress to be released. The relationship between ancient fault systems and the Tweed Shield Volcano has also been discussed and it is suggested here that the volcanic activity was associated with renewed faulting on the Great Moreton Fault System during the Cenozoic. The geomorphology and drainage patterns of southeast Queensland have been compared with expected morphological characteristics found at passive and other tectonic settings, both in Australia and globally. Of note are the comparisons with the East Brazilian Highlands, the Gulf of Mexico and the Blue Ridge Escarpment, for example. In conclusion, the results of the study clearly show that, although the region is described as a passive margin, its complex, past geological history and present compressive stress regime provide a more intricate and varied landscape than would be expected along typical passive continental margins. The literature review provides background to the subject and discusses previous work and methods, whilst the findings are presented in three peer-reviewed, published papers. The methods, hypotheses, suggestions and evidence are discussed at length in the final chapter.
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This paper explores a method of comparative analysis and classification of data through perceived design affordances. Included is discussion about the musical potential of data forms that are derived through eco-structural analysis of musical features inherent in audio recordings of natural sounds. A system of classification of these forms is proposed based on their structural contours. The classifications include four primitive types; steady, iterative, unstable and impulse. The classification extends previous taxonomies used to describe the gestural morphology of sound. The methods presented are used to provide compositional support for eco-structuralism.
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This paper investigates the robust H∞ control for Takagi-Sugeno (T-S) fuzzy systems with interval time-varying delay. By employing a new and tighter integral inequality and constructing an appropriate type of Lyapunov functional, delay-dependent stability criteria are derived for the control problem. Because neither any model transformation nor free weighting matrices are employed in our theoretical derivation, the developed stability criteria significantly improve and simplify the existing stability conditions. Also, the maximum allowable upper delay bound and controller feedback gains can be obtained simultaneously from the developed approach by solving a constrained convex optimization problem. Numerical examples are given to demonstrate the effectiveness of the proposed methods.
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This paper describes protection and control of a microgrid with converter interfaced micro sources. The proposed protection and control scheme consider both grid connected and autonomous operation of the microgrid. A protection scheme, capable of detecting faults effectively in both grid connected and islanded operations is proposed. The main challenge of the protection, due to current limiting state of the converters is overcome by using admittance relays. The relays operate according to the inverse time characteristic based on measured admittance of the line. The proposed scheme isolates the fault from both sides, while downstream side of the microgrid operates in islanding condition. Moreover faults can be detected in autonomous operation. In grid connected mode distributed generators (DG) supply the rated power while in absence of the grid, DGs share the entire power requirement proportional to rating based on output voltage angle droop control. The protection scheme ensures minimum load shedding with isolating the faulted network and DG control provides a smooth islanding and resynchronization operation. The efficacy of coordinated control and protection scheme has been validated through simulation for various operating conditions.
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Background/Aims: In an investigation of the functional impact of amblyopia on children, the fine motor skills, perceived self-esteem and eye movements of amblyopic children were compared with that of age-matched controls. The influence of amblyogenic condition or treatment factors that might predict any decrement in outcome measures was investigated. The relationship between indirect measures of eye movements that are used clinically and eye movement characteristics recorded during reading was examined and the relevance of proficiency in fine motor skills to performance on standardised educational tests was explored in a sub-group of the control children. Methods: Children with amblyopia (n=82; age 8.2 ± 1.3 years) from differing causes (infantile esotropia n=17, acquired strabismus n=28, anisometropia n=15, mixed n=13 and deprivation n=9), and a control group of children (n=106; age 9.5 ± 1.2 years) participated in this study. Measures of visual function included monocular logMAR visual acuity (VA) and stereopsis assessed with the Randot Preschool Stereoacuity test, while fine motor skills were measured using the Visual-Motor Control (VMC) and Upper Limb Speed and Dexterity (ULSD) subtests of the Brunicks-Oseretsky Test of Motor Proficiency. Perceived self esteem was assessed for those children from grade 3 school level with the Harter Self Perception Profile for Children and for those in younger grades (preschool to grade 2) with the Pictorial Scale of Perceived Competence and Acceptance for Young Children. A clinical measure of eye movements was made with the Developmental Eye Movement (DEM) test for those children aged eight years and above. For appropriate case-control comparison of data, the results from amblyopic children were compared with age-matched sub-samples drawn from the group of children with normal vision who completed the tests. Eye movements during reading for comprehension were recorded by the Visagraph infra-red recording system and results of standardised tests of educational performance were also obtained for a sub-set of the control group. Results Amblyopic children (n=82; age 8.2 ± 1.7 years) performed significantly poorer than age-matched control children (n=37; age 8.3 ± 1.3 years) on 9 of 16 fine motor skills sub-items and for the overall age-standardised scores for both VMC and ULSD items (p<0.05); differences were most evident on timed manual dexterity tasks. The underlying aetiology of amblyopia and level of stereoacuity significantly affected fine motor skill performance on both items. However, when examined in a multiple regression model that took into account the inter-correlation between visual characteristics, poorer fine motor skills performance was only associated with strabismus (F1,75 = 5.428; p =0. 022), and not with the level of stereoacuity, refractive error or visual acuity in either eye. Amblyopic children from grade 3 school level and above (n=47; age 9.2 ± 1.3 years), particularly those with acquired strabismus, had significantly lower social acceptance scores than age-matched control children (n=52; age 9.4 ± 0.5 years) (F(5,93) = 3.14; p = 0.012). However, the scores of the amblyopic children were not significantly different to controls for other areas related to self-esteem, including scholastic competence, physical appearance, athletic competence, behavioural conduct and global self worth. A lower social acceptance score was independently associated with a history of treatment with patching but not with a history of strabismus or wearing glasses. Amblyopic children from pre-school to grade 2 school level (n=29; age = 6.6 ± 0.6 years) had similar self-perception scores to their age-matched peers (n=20; age = 6.4 ± 0.5 years). There were no significant differences between the amblyopic (n=39; age 9.1 ± 0.9 years) and age-matched control (n = 42; age = 9.3 ± 0.38 years) groups for any of the DEM outcome measures (Vertical Time, Horizontal Time, Number of Errors and Ratio (Horizontal time/Vertical time)). Performance on the DEM did not significantly relate to measures of VA in either eye, level of binocular function, history of strabismus or refractive error. Developmental Eye Movement test outcome measures Horizontal Time and Vertical Time were significantly correlated with reading rates measured by the Visagraph for both reading for comprehension and naming numbers (r>0.5). Some moderate correlations were also seen between the DEM Ratio and word reading rates as recorded by Visagraph (r=0.37). In children with normal vision, academic scores in mathematics, spelling and reading were associated with measures of fine motor skills. Strongest effect sizes were seen with the timed manual dexterity domain, Upper Limb Speed and Dexterity. Conclusions Amblyopia may have a negative impact on a child’s fine motor skills and an older child’s sense of acceptance by their peers may be influenced by treatment that includes eye patching. Clinical measures of eye movements were not affected in amblyopic children. A number of the outcome measures of the DEM are associated with objective recordings of reading rates, supporting its clinical use for identification of children with slower reading rates. In children with normal vision, proficiency on clinical measures of fine motor skill are associated with outcomes on standardised measures of educational performance. Scores on timed manual dexterity tasks had the strongest association with educational performance. Collectively, the results of this study indicate that, in addition to the reduction in visual acuity and binocular function that define the condition, amblyopes have functional impairment in childhood development skills that underlie proficiency in everyday activities. The study provides support for strategies aimed at early identification and remediation of amblyopia and the co-morbidities that arise from abnormal visual neurodevelopment.
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Over the past decade, plants have been used as expression hosts for the production of pharmaceutically important and commercially valuable proteins. Plants offer many advantages over other expression systems such as lower production costs, rapid scale up of production, similar post-translational modification as animals and the low likelihood of contamination with animal pathogens, microbial toxins or oncogenic sequences. However, improving recombinant protein yield remains one of the greatest challenges to molecular farming. In-Plant Activation (InPAct) is a newly developed technology that offers activatable and high-level expression of heterologous proteins in plants. InPAct vectors contain the geminivirus cis elements essential for rolling circle replication (RCR) and are arranged such that the gene of interest is only expressed in the presence of the cognate viral replication-associated protein (Rep). The expression of Rep in planta may be controlled by a tissue-specific, developmentally regulated or chemically inducible promoter such that heterologous protein accumulation can be spatially and temporally controlled. One of the challenges for the successful exploitation of InPAct technology is the control of Rep expression as even very low levels of this protein can reduce transformation efficiency, cause abnormal phenotypes and premature activation of the InPAct vector in regenerated plants. Tight regulation over transgene expression is also essential if expressing cytotoxic products. Unfortunately, many tissue-specific and inducible promoters are unsuitable for controlling expression of Rep due to low basal activity in the absence of inducer or in tissues other than the target tissue. This PhD aimed to control Rep activity through the production of single chain variable fragments (scFvs) specific to the motif III of Tobacco yellow dwarf virus (TbYDV) Rep. Due to the important role played by the conserved motif III in the RCR, it was postulated that such scFvs can be used to neutralise the activity of the low amount of Rep expressed from a “leaky” inducible promoter, thus preventing activation of the TbYDV-based InPAct vector until intentional induction. Such scFvs could also offer the potential to confer partial or complete resistance to TbYDV, and possibly heterologous viruses as motif III is conserved between geminiviruses. Studies were first undertaken to determine the levels of TbYDV Rep and TbYDV replication-associated protein A (RepA) required for optimal transgene expression from a TbYDV-based InPAct vector. Transient assays in a non-regenerable Nicotiana tabacum (NT-1) cell line were undertaken using a TbYDV-based InPAct vector containing the uidA reporter gene (encoding GUS) in combination with TbYDV Rep and RepA under the control of promoters with high (CaMV 35S) or low (Banana bunchy top virus DNA-R, BT1) activity. The replication enhancer protein of Tomato leaf curl begomovirus (ToLCV), REn, was also used in some co-bombardment experiments to examine whether RepA could be substituted by a replication enhancer from another geminivirus genus. GUS expression was observed both quantitatively and qualitatively by fluorometric and histochemical assays, respectively. GUS expression from the TbYDV-based InPAct vector was found to be greater when Rep was expected to be expressed at low levels (BT1 promoter) rather than high levels (35S promoter). GUS expression was further enhanced when Rep and RepA were co-bombarded with a low ratio of Rep to RepA. Substituting TbYDV RepA with ToLCV REn also enhanced GUS expression but more importantly highest GUS expression was observed when cells were co-transformed with expression vectors directing low levels of Rep and high levels of RepA irrespective of the level of REn. In this case, GUS expression was approximately 74-fold higher than that from a non-replicating vector. The use of different terminators, namely CaMV 35S and Nos terminators, in InPAct vectors was found to influence GUS expression. In the presence of Rep, GUS expression was greater using pInPActGUS-Nos rather than pInPActGUS-35S. The only instance of GUS expression being greater from vectors containing the 35S terminator was when comparing expression from cells transformed with Rep, RepA and REnexpressing vectors and either non-replicating vectors, p35SGS-Nos or p35SGS-35S. This difference was most likely caused by an interaction of viral replication proteins with each other and the terminators. These results indicated that (i) the level of replication associated proteins is critical to high transgene expression, (ii) the choice of terminator within the InPAct vector may affect expression levels and (iii) very low levels of Rep can activate InPAct vectors hence controlling its activity is critical. Prior to generating recombinant scFvs, a recombinant TbYDV Rep was produced in E. coli to act as a control to enable the screening for Rep-specific antibodies. A bacterial expression vector was constructed to express recombinant TbYDV Rep with an Nterminal His-tag (N-His-Rep). Despite investigating several purification techniques including Ni-NTA, anion exchange, hydrophobic interaction and size exclusion chromatography, N-His-Rep could only be partially purified using a Ni-NTA column under native conditions. Although it was not certain that this recombinant N-His-Rep had the same conformation as the native TbYDV Rep and was functional, results from an electromobility shift assay (EMSA) showed that N-His-Rep was able to interact with the TbYDV LIR and was, therefore, possibly functional. Two hybridoma cell lines from mice, immunised with a synthetic peptide containing the TbYDV Rep motif III amino acid sequence, were generated by GenScript (USA). Monoclonal antibodies secreted by the two hybridoma cell lines were first screened against denatured N-His-Rep in Western analysis. After demonstrating their ability to bind N-His-Rep, two scFvs (scFv1 and scFv2) were generated using a PCR-based approach. Whereas the variable heavy chain (VH) from both cell lines could be amplified, only the variable light chain (VL) from cell line 2 was amplified. As a result, scFv1 contained VH and VL from cell line 1, whereas scFv2 contained VH from cell line 2 and VL from cell line 1. Both scFvs were first expressed in E. coli in order to evaluate their affinity to the recombinant TbYDV N-His-Rep. The preliminary results demonstrated that both scFvs were able to bind to the denatured N-His-Rep. However, EMSAs revealed that only scFv2 was able to bind to native N-His-Rep and prevent it from interacting with the TbYDV LIR. Each scFv was cloned into plant expression vectors and co-bombarded into NT-1 cells with the TbYDV-based InPAct GUS expression vector and pBT1-Rep to examine whether the scFvs could prevent Rep from mediating RCR. Although it was expected that the addition of the scFvs would result in decreased GUS expression, GUS expression was found to slightly increase. This increase was even more pronounced when the scFvs were targeted to the cell nucleus by the inclusion of the Simian virus 40 large T antigen (SV40) nuclear localisation signal (NLS). It was postulated that the scFvs were binding to a proportion of Rep, leaving a small amount available to mediate RCR. The outcomes of this project provide evidence that very high levels of recombinant protein can theoretically be expressed using InPAct vectors with judicious selection and control of viral replication proteins. However, the question of whether the scFvs generated in this project have sufficient affinity for TbYDV Rep to prevent its activity in a stably transformed plant remains unknown. It may be that other scFvs with different combinations of VH and VL may have greater affinity for TbYDV Rep. Such scFvs, when expressed at high levels in planta, might also confer resistance to TbYDV and possibly heterologous geminiviruses.
Resumo:
Background: There are innumerable diabetes studies that have investigated associations between risk factors, protective factors, and health outcomes; however, these individual predictors are part of a complex network of interacting forces. Moreover, there is little awareness about resilience or its importance in chronic disease in adulthood, especially diabetes. Thus, this is the first study to: (1) extensively investigate the relationships among a host of predictors and multiple adaptive outcomes; and (2) conceptualise a resilience model among people with diabetes. Methods: This cross-sectional study was divided into two research studies. Study One was to translate two diabetes-specific instruments (Problem Areas In Diabetes, PAID; Diabetes Coping Measure, DCM) into a Chinese version and to examine their psychometric properties for use in Study Two in a convenience sample of 205 outpatients with type 2 diabetes. In Study Two, an integrated theoretical model is developed and evaluated using the structural equation modelling (SEM) technique. A self-administered questionnaire was completed by 345 people with type 2 diabetes from the endocrine outpatient departments of three hospitals in Taiwan. Results: Confirmatory factor analyses confirmed a one-factor structure of the PAID-C which was similar to the original version of the PAID. Strong content validity of the PAID-C was demonstrated. The PAID-C was associated with HbA1c and diabetes self-care behaviours, confirming satisfactory criterion validity. There was a moderate relationship between the PAID-C and the Perceived Stress Scale, supporting satisfactory convergent validity. The PAID-C also demonstrated satisfactory stability and high internal consistency. A four-factor structure and strong content validity of the DCM-C was confirmed. Criterion validity demonstrated that the DCM-C was significantly associated with HbA1c and diabetes self-care behaviours. There was a statistical correlation between the DCM-C and the Revised Ways of Coping Checklist, suggesting satisfactory convergent validity. Test-retest reliability demonstrated satisfactory stability of the DCM-C. The total scale of the DCM-C showed adequate internal consistency. Age, duration of diabetes, diabetes symptoms, diabetes distress, physical activity, coping strategies, and social support were the most consistent factors associated with adaptive outcomes in adults with diabetes. Resilience was positively associated with coping strategies, social support, health-related quality of life, and diabetes self-care behaviours. Results of the structural equation modelling revealed protective factors had a significant direct effect on adaptive outcomes; however, the construct of risk factors was not significantly related to adaptive outcomes. Moreover, resilience can moderate the relationships among protective factors and adaptive outcomes, but there were no interaction effects of risk factors and resilience on adaptive outcomes. Conclusion: This study contributes to an understanding of how risk factors and protective factors work together to influence adaptive outcomes in blood sugar control, health-related quality of life, and diabetes self-care behaviours. Additionally, resilience is a positive personality characteristic and may be importantly involved in the adjustment process among people living with type 2 diabetes.
Resumo:
There is considerable evidence that working memory impairment is a common feature of schizophrenia. The present study assessed working memory and executive function in 54 participants with schizophrenia, and a group of 54 normal controls matched to the patients on age, gender and estimated premorbid IQ, using traditional and newer measures of executive function and two dual tasks—Telephone Search with Counting and the Memory Span and Tracking Task. Results indicated that participants with schizophrenia were significantly impaired on all standardised measures of executive function with the exception of a composite measure of the Trail Making Test. Results for the dual task measures demonstrated that while the participants with schizophrenia were unimpaired on immediate digit span recall over a 2-min period, they recalled fewer digit strings and performed more poorly on a tracking task (box-crossing task) compared with controls. In addition, participants with schizophrenia performed more poorly on the tracking task when they were required to simultaneously recall digits strings than when they performed this task alone. Contrary to expectation, results of the telephone search task under dual conditions were not significantly different between groups. These results may reflect the insufficient complexity of the tone-counting task as an interference task. Overall, the present study showed that participants with schizophrenia appear to have a restricted impairment of their working memory system that is evident in tasks in which the visuospatial sketchpad slave system requires central executive control.