Novel conserved amino acid variations of papaya cystatin (PC) were investigated by amino acid substitutions using oryzacystatin-I (OCI) as a model plant cystatin for comparison. These amino acid residues in the conserved motifs are involved in binding with cysteine proteases, these include the GG (Gly-Gly) in the N-terminal region for both OCI and PC, the (Q)QVVAG (Gln-Val-Val-Ala-Gly) motif for OCI and (Q)AVVEG (Ala-Val-Val-Glu-Gly) motif for PC in the first inhibitory loop, and the PW (Pro-Trp) motif for OCI and LW (Leu-Trp) motif for PC in the second inhibitory loop. Recombinant OCI and PC mutant proteins were expressed in Escherichia coli and were tested for altered inhibitory activity against commercial cysteine proteases (papain and cathepsin L) and extracts from Colorado potato beetle (Leptinotarsa decemlineata) larvae, from banana weevil larvae (Cosmopolites sordidus) and tobacco leaf extracts (Nicotiana benthamiana). In all tests higher amounts of PC had to be used to obtain similar inhibition levels as OCI. Changing the amino acid Q at position 52 to E in OCI in the first inhibitory loop, had lowered the Ki value of the mutant against the commercial proteases. Concurrently the same amino acid string (EQ) in PC had resulted in a significantly decreased Ki value compared to PC wild-type and other mutants. All other OCI mutants were less efficient than the wild-type OCI, whereas all PC first inhibitory loop mutants had improved inhibitory activity against protease activity with the highest improvement against the protease extracts was found for the substitution of E with A at position 55. This study has shown the importance of the three conserved motifs and that it is possible to improve the binding capacity of a plant cystatins to cysteine protease activity by amino acid substitution using site-directed mutagenesis. By mutating individual amino acid residues in the first binding loop of the relatively “weak” papaya cystatin to amino acid residues found in OCI caused a significant improvement in inhibitory potency of PC. Copyright / Dissertation (MSc)--University of Pretoria, 2011. / Plant Science / unrestricted

This diploma thesis deals with communication in kindergarten in selected communicative situations. The first part defines basic theoretical terms and, based on previous research, summarizes findings about pre-school education and mental and language development in pre- school children. The thesis also explains factors that influence language acquisition, child directed speech and pedagogical communication. The second part's main focus lies in describing communicative situations and in qualitative analysis of the acquired data - video recordings of selected communicative situation. The pocess of recording took place after previous agreement with two kindergartens. The recordings were transcribed according to the modified manual for the DIALOG database. The analysis is focused on the nature of the pedagogical communication in kindergarten. Our findings were compared to previous research results. Keywords communication, pedagogical communication, language acquisition, child directed speech, kindergarten

Counterfeit consumption is becoming widespread, developing into a problem of international significance. In an attempt to develop a refined understanding of the motivations and decision-making processes of consumers' deliberate counterfeit-consumption behavior, this empirical study not only integrates the theory of planned behavior and insights from self-regulatory theories, but also extends these theories by re-conceptualizing the relationships among key constructs with the inclusion of action desire. This research also introduces and integrates a new construct, namely consumers' Perceived Counterfeit Detection (PCD) by important others.Using a combination of qualitative and quantitative methods consisting of in-depth interviews and a self-administered paper questionnaire, this research empirically tested a proposed goal-striving model for deliberate counterfeit-consumption behavior, using structural equations modeling. Results demonstrated PCD's existence and supported a refined goal-striving model, based upon which effective strategies to decrease consumer's counterfeit consumption are discussed.

In South Africa, the trend has been towards the decentralising of authority and giving greater powers to schools. This implies the active involvement of the school governing body members, the school management team members and the Level One educators in the day-to-day functioning of the school. Whilst some schools have welcomed this change and seen this devolution of power as an added advantage contributing to the efficacy of schools, some schools have struggled, while others have failed dismally to cope with this added responsibility of managing their own resources. The researcher, being an educator involved in this transition, has been intrigued by the success of some self-managing schools and the failure of others. The following words of Oliver Wendell Holmes have inspired the investigator to conduct this research to ascertain the reasons for the success of certain schools and the failure of others: “What lies behind us and what lies before us are tiny matters compared to what lies within us.” Since the performance of schools is normally ranked according to their matriculation pass rates, this research focused on six self-managing schools in the Port Elizabeth District. These schools were selected on the basis of their matriculation performances in 2008. Two of the schools had achieved an above 85 percent matriculation pass rate (Category A); two obtained a 50 percent pass rate (Category B); while the other two schools obtained matriculation pass rates of between 25 percent and 30 percent (Category C). Although the research relied strongly on a qualitative method of inquiry, because of the nature of the data and the need to obtain a better understanding of the feelings, attitude and perceptions of those in management positions, governing body positions and members of the personnel, multiple instruments for data collection such as a literature review and the study of school documents and questionnaires, were used. This inclusion of these multiple instruments for obtaining information, which is also called triangulation, contributed to the reliability and validity of the empirical investigation. Although the data was presented in the form of tables and graphs, vi which alludes to a quantitative approach, a narrative interpretation of the findings, which is associated with a qualitative method of inquiry, was also presented. An analysis of the findings revealed that there was a great amount of involvement of the principals, school management team members and Level One educators in Category A schools, which had a matriculation pass rate of above 85%, in the following change management strategies: the establishment of a vision, planning and organising, teamwork, training and development, communication, praise and incentives, and evaluation and feedback. There was also a great contribution by the school governing body members. However, the study revealed that Category B and Category C schools did not effectively use communication, praise and incentives and evaluation and feedback as change management strategies. It was evident from the empirical investigation that in Category B and C schools, the school governing body members were less, or not involved in the usage of the change management strategies applied. According to the results based on the six-facet model of Emotional Intelligence, Category A schools obtained 80% and above in the competencies relating to the self and social domain. Category B schools fluctuated between 50 percent and 80 percent in both dimensions, while Category C schools fluctuated between 30 percent and 80 percent. The findings suggest that there should be greater involvement of all stakeholders, especially the school governing body members, in the establishment of a vision, planning and organising, communication, teamwork, training and development, praise and incentives and the provision of feedback and evaluation for self-managing schools to become effective and improve their performances. Professional development, especially in the areas pertaining to Emotional Intelligence competencies such as personal and interpersonal skills, should be prioritised. This thesis proposes that, on the basis of the investigation carried out at the six participating schools on the influence of Emotional Intelligence on the change management strategies adopted in self-managing schools, schools use strategies such as the establishment of a vision, planning and organising, communication, teamwork, training and development, praise and incentives and evaluation and feedback to ameliorate the damaging impact of change. There should be continuous training and development at schools, especially for the school governing body members, who are elected every three years. The newly elected school governing body members need to develop the necessary skills to effectively contribute to the efficacy of schools. The human resource is the vital ingredient in schools, thus for any progression or improvement, all stakeholders need to be informed, trained and enriched. Since Emotional Intelligence can be learned, professional development in the area of Emotional Intelligence should be given more thought and included in training courses, focusing on the development of personal and interpersonal skills. The Emotional Intelligence training should be conducted as part of teacher training courses as well as on an annual basis for educators already in the teaching system, since Emotional Intelligence can influence the use of change management strategies at schools and enhance the efficacy of the functioning of self-managing schools.

The aim of this thesis is to investigate aspects of molecular evolution and enzyme engineering using the experimental evolution of Tobacco Etch Virus cysteine protease (TEV) as a model. I map key features of the local fitness landscape and characterise how they affect details of enzyme evolution. In order to investigate the evolution of core active site machinery, I mutated the nucleophile of TEV to serine. The differing chemical properties of oxygen and sulphur force the enzyme into a fitness valley with a >104-fold activity reduction. Nevertheless, directed evolution was able to recover function, resulting in an enzyme able to utilise either nucleophile. High-throughput screening and sequencing revealed how the array of possible beneficial mutations changes as the enzyme evolves. Potential adaptive mutations are abundant at each step along the evolutionary trajectory, enriched around the active site periphery. It is currently unclear how seemingly neutral mutations affect further adaptive evolution. I used high-throughput directed evolution to accumulate neutral variation in large, evolving enzyme populations and deep sequencing to reconstruct the complex evolutionary dynamics within the lineages. Specifically I was able to observe the emergence of robust enzymes with improved mutation tolerance whose descendants overtake later populations. Lastly, I investigate how evolvability towards new substrate specificities changed along these neutral lineages, dissecting the different determinants of immediate and long-term evolvability. Results demonstrate the utility of evolutionary understanding to protease engineering. Together, these experiments forward our understanding of the molecular details of both fundamental evolution and enzyme engineering.

Aim: This paper is a report on a study that explored the professional development of intensive care unit nurses in journal clubs. Background: Evidence-based practice is important in nursing care (Krom, Batten, & Bautista, 2010). However few nurses feel comfortable using evidence to guide their practice (Pravikoff, Tanner, & Pierce, 2005). Journal clubs are a way to establish science as conversation (Wright, 2004) and foster knowledge translation for evidence-based nursing practice (Goodfellow, 2004). Methods: Monthly journal club meetings were held with the participation of a total of 71 healthcare professionals (65 nurses, 2 physicians, 2 pharmacists, 1 physiotherapist, and 1 respiratory therapist), who worked in two intensive care units of an Ontario hospital. After six months of meetings, 21 individual interviews were conducted with nurses, physicians, pharmacists, and nurse educators. Additional data collection included two focus groups, surveys, a review of staff meeting minutes, and researcher field notes. Findings: Journal clubs provided nurses with incentive to read research articles, improved nurses’ confidence in reading research, created a community of peers who worked collaboratively to improve clinical practice, provided a structure for nurses to reflect-on-practice, and led to reported changes in clinical practice. However, the data suggests that any gains in competence of nurses with the critical appraisal of research articles were probably modest. Barriers to participating in journal clubs and evidence-based practice are also identified. Conclusion: Journal clubs can foster knowledge translation and evidence-based practice through creating a community of practice and by providing nurses with motivation, structure, and confidence to read research articles. However, nurses reported a lack of critical appraisal skills and uncertainty about how to implement evidence into practice. Journal clubs may have a greater impact when implemented alongside other knowledge translation strategies such as working with clinical nurse specialists in order to enhance evidence-based practice.

CHOICES, the career planning computer program, was evaluated by interviewing 35 grade 11 and 12 students. Using the critical incident technique, reports were elicited of what facilitated or hindered their career planning. These collected incidents were categorized by similarity to provide counsellors and others with a map of exactly what the program does to help or hinder career planning. This map potentially enables counsellors to capitalize on benefits and to minimize possible detriments. Secondly, this map was qualitatively compared to a similar evaluation of the Self-Directed Search. Overall, it was found that the two interventions have differing advantages and disadvantages. CHOICES stresses reality constraints, specificity and extrinsic work features. The SDS underlines self-awareness and an understanding of the matching process. It seems that CHOICES is more appropriate for planning and specific decisions regarding options while the SDS tends to focus on general exploration and decisions regarding fields. The writer wishes to express his appreciation to the following persons: To Dr. Larry Cochran, research supervisor, whose patience, availability and countless hours over six years made this study possible. To Dr. Marv Westwood, whose continual support especially for the clinical comprehensive, was greatly appreciated. To Dr. Bill Borgen, whose encouragement at the appropriate time enabled me to perservere. To Dr. Norm Amundson, whose inspired synthesis made the oral presentation smoother. To Dr. Ron MacGregor, whose suggestions were appreciated. To the students, to the staff of Matthew McNair Senior Secondary and to the counsellor Mike Casselman, whose co-operation and support made this study possible. To Mrs. Irma Eichler, for her devoted time and effort in typing this dissertation. And finally, to my parents Rene and Lucille, to my sisters Louise and Anita, and to my brother Jean, for their continual support and encouragement. / Education, Faculty of / Educational and Counselling Psychology, and Special Education (ECPS), Department of / Graduate

Dans un contexte professionnel en perpétuelle mutation, les salariés ont la nécessité de se former régulièrement, et ce quel que soit leur niveau de formation initial. Nous avons coutume de lire et d’entendre que le taux d’abandon et le taux d’échec des apprenants dans un dispositif de FOAD est supérieur à celui observé dans la formation présentielle. Ces défections ou revers peuvent être la résultante de divers facteurs dont: l'isolement de l'apprenant, un défaut d'ergonomie de l'environnement d’apprentissage (pédagogique, technologique) ou bien le manque d'autonomie de l'apprenant. Pour passer d’un système fondé sur la transmission du savoir (présentiel) à un système fondé sur l’appropriation et la création de connaissances (FOAD), il faut consentir des efforts particuliers, aussi, notre objectif est de découvrir : comment s’y prennent ceux qui vont jusqu’au bout et réussissent leur formation ? Pour répondre à cette question, nous allons investiguer les styles d’apprentissage des apprenants, qui induisent les méthodologies et stratégies d’apprentissage, ainsi que leur dimension auto-formative qui est une composante importante de la FOAD, due à l’éclatement des unités de temps, de lieu, d’action. Ainsi, nous souhaitons découvrir quels sont les styles d’apprentissage des participants d’une part, en essayant de voir quels sont ceux qui éventuellement dominent, et d’autre part, définir quelle est la capacité d’auto-apprentissage des participants dans le processus de formation ouverte et à distance. La FOAD suppose un ancrage fort dans l’humain, notamment les dimensions psycho-affectives et cognitives. Tous les participants ayant réussi leur formation, nous espérons découvrir chez eux des traits individuels, des dominantes ou constantes comportementales qui pourraient peut-être l’expliquer. / In a constantly changing professional environment, employees need to train regularly, regardless of their level of initial training. We usually read and hear that the dropout and failure rate of learners in an open distance learning device is higher than that observed in the classroom training face to face. These defections or setbacks may be the result of various factors including: the isolation of the learner, poor ergonomics of learning environments (educational technology) or the lack of learner autonomy. To move from a system based on knowledge transfer (face to face) to a system based on ownership and knowledge creation (Open Distance Learning), we must make special efforts, however, our goal is to discover how they managed to complete their education and succeed? To answer this question, we will investigate the learning styles of learners, which induce the methodologies and learning strategies and their self-formative dimension that is an important component of distance learning, due to the breakdown of unit’s time, place, and action. Thus, we would discover what are the learning styles of participants on the one hand, trying to see which ones may dominate the other hand, define what is the capacity for self-learning participants in the process of open and distance learning.ODL requires a strong anchoring in the human, including dimensions psycho-emotional and cognitive. All participants who successfully completed their training, we hope to discover in their individual traits, the dominant behavioral or constants that could possibly explain.

Autism Spectrum disorder (ASD) is a wide ranging developmental condition commonly recognised by impairments in social interaction, communication, repetitive behaviour and restricted interests. Deficits in imitation, sensory processing and motor control are also extensive, but conflicting findings mean a clear picture regarding the true nature of these is yet to be established. The aim of this thesis was to further investigate imitation with a specific focus on the effect of goals. It has been suggested that imitation occurs along two routes; a direct route for the imitation of meaningless actions i.e. goal-less, and a semantic route for meaningful or goal-directed actions. There is evidence to suggest that while individuals with ASD may have an impairment in goal-less imitation, goal-directed imitation is unaffected (Hamilton, Brindley & Frith, 2007). The experiments in the present work were based on the hypothesis that impaired goal-less imitation in ASD may be due to a problem with sensory motor integration in the direct visuomotor pathway for imitation (Tessari and Rumiati, 2004).The first experiment was conducted to ascertain whether movement kinematics differed between imitation during goal-directed and goal-less hand movements in a neurotypical control group. Participants observed and imitated hand movements of different speeds, while their movement was recorded with a motion sensor. Movement was modulated between the different speeds in the goal-less, but not goal-directed trials. These findings support the dual route model where visuomotor mapping occurs via the direct pathway during goal-less imitation but during goal-directed imitation there is greater reliance on representations of actions from long term memory.In experiment two, the same paradigm was employed, including additional movement types, and a group of adult individuals diagnosed with an ASD was compared to a control group. It was predicted that, unlike the control group, the ASD group would fail to modulate their movement in the goal-less condition, due to a disruption in the direct pathway. Eye movements were also recorded in this experiment, to ascertain if differences in gaze position or eye movements might influence the ability of the ASD group to imitate goal-less actions. The ASD group did not modulate their movement for goal-less imitation. In addition, the ASD group exhibited more goal-directed gaze, and less gaze toward the hand, than the control group. The ASD group also failed to maintain pursuit of the hand, which may have influenced the amount of information collected regarding the movement.The third study extended the investigation by introducing a training phase halfway through the experiment. Based on eye movement findings, it was hypothesised that drawing attention to the hand might increase the importance of the hand in goal-less trials, subsequently leading to increased tracking, and improved imitation. Perspective was also investigated to determine whether observing the action from an egocentric perspective would improve imitation. Movement modulation was not improved for either the post training or the egocentric condition. However, both training and egocentric viewing resulted in faster performance of movements while training also reduced the amount of movement variability; suggesting that these conditions made imitation easier. Findings strongly support the hypothesis that people with ASD are impaired in goal-less but not goal-directed imitation. These results are discussed in terms of bottom-up and top-down influences on imitation. The observed eye movement differences between the ASD and control group suggest eye movement abnormalities, and the finding that egocentric observation facilitates imitation indicates problems with visuospatial transformation during allocentric imitation. Finally, failure to pursue may be due to lack of attention to the hand movement, evidenced by no increase in pursuit after training. This thesis concludes with the suggestion that impaired goal-less imitation in ASD may result primarily from bottom-up low level visual processing and oculomotor control problems, combined with a high level attentional mechanism. Future studies need to address whether these are the primary causes of imitation difficulties, or whether there is a fundamental sensory motor integration deficit in ASD.

Optimisation is one of the most important and challenging part of any engineering design. In real world design problems one faces multiobjective optimisation under constraints. The optimal solution in these cases is not unique because the objectives can contradict each other. In such cases, a set of optimal solutions which forms a Pareto frontier in the objective space is considered. There are many algorithms to generate the Pareto frontier. However, only a few of them are potentially capable of providing an evenly distributed set of the solutions. Such a property is especially important in real-life design because a decision maker is usually able to analyse only a very limited quantity of solutions. This thesis consists of two main parts. At first, it develops and gives the detailed description of two different algorithms that are able to generate an evenly distributed Pareto set in a general formulation. One is a classical approach and called Directed Search Domain (DSD) and the other, the cylindrical constraint evolutionary algorithm (CCEA), is a hybrid population based method. The efficiency of the algorithms are demonstrated by a number of challenging test cases and the comparisons with the results of the other existing methods. It is shown that the proposed methods are successful in generating the Pareto solutions even when some existing methods fail. In real world design problems, deterministic approaches cannot provide a reliable solution as in the event of uncertainty, deterministic optimal solution would be infeasible in many instances. Therefore a solution less sensitive to problem perturbation is desirable. This leads to the robust solution which is the focus of the second part of the thesis. In the literature, there are some techniques tailored for robust optimisation. However, most of them are either computationally expensive or do not systematically articulate the designer preferences into a robust solution. In this thesis, by introducing a measure for robustness in multiobjective context, a tunable robust function (TRF) is presented. Including the TRF in the problem formulation, it is demonstrated that the desirable robust solution based on designer preferences can be obtained. This not only provides the robust solution but also gives a control over the robustness level. The method is efficient as it only increases the dimension of the problem by one irrespective of the dimension of the original problem.