Improved production of industrially relevant recombinant proteins: application of multi-factorial design of experiments and scalable process modelling

The work described in this thesis focuses on the use of a design-of-experiments approach in a multi-well mini-bioreactor to enable the rapid establishments of high yielding production phase conditions in yeast, which is an increasingly popular host system in both academic and industrial laboratories. Using green fluorescent protein secreted from the yeast, Pichia pastoris, a scalable predictive model of protein yield per cell was derived from 13 sets of conditions each with three factors (temperature, pH and dissolved oxygen) at 3 levels and was directly transferable to a 7 L bioreactor. This was in clear contrast to the situation in shake flasks, where the process parameters cannot be tightly controlled. By further optimisating both the accumulation of cell density in batch and improving the fed-batch induction regime, additional yield improvement was found to be additive to the per cell yield of the model. A separate study also demonstrated that improving biomass improved product yield in a second yeast species, Saccharomyces cerevisiae. Investigations of cell wall hydrophobicity in high cell density P. pastoris cultures indicated that cell wall hydrophobin (protein) compositional changes with growth phase becoming more hydrophobic in log growth than in lag or stationary phases. This is possibly due to an increased occurrence of proteins associated with cell division. Finally, the modelling approach was validated in mammalian cells, showing its flexibility and robustness. In summary, the strategy presented in this thesis has the benefit of reducing process development time in recombinant protein production, directly from bench to bioreactor.

The influence of twist drill design on rigidity and chip disposal

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Online-questionnaire design:establishing guidelines and evaluating existing eupport

As a new medium for questionnaire delivery, the internet has the potential to revolutionise the survey process. Online (web-based) questionnaires provide several advantages over traditional survey methods in terms of cost, speed, appearance, flexibility, functionality, and usability [1, 2]. For instance, delivery is faster, responses are received more quickly, and data collection can be automated or accelerated [1- 3]. Online-questionnaires can also provide many capabilities not found in traditional paper-based questionnaires: they can include pop-up instructions and error messages; they can incorporate links; and it is possible to encode difficult skip patterns making such patterns virtually invisible to respondents. Like many new technologies, however, online-questionnaires face criticism despite their advantages. Typically, such criticisms focus on the vulnerability of online-questionnaires to the four standard survey error types: namely, coverage, non-response, sampling, and measurement errors. Although, like all survey errors, coverage error (“the result of not allowing all members of the survey population to have an equal or nonzero chance of being sampled for participation in a survey” [2, pg. 9]) also affects traditional survey methods, it is currently exacerbated in online-questionnaires as a result of the digital divide. That said, many developed countries have reported substantial increases in computer and internet access and/or are targeting this as part of their immediate infrastructural development [4, 5]. Indicating that familiarity with information technologies is increasing, these trends suggest that coverage error will rapidly diminish to an acceptable level (for the developed world at least) in the near future, and in so doing, positively reinforce the advantages of online-questionnaire delivery. The second error type – the non-response error – occurs when individuals fail to respond to the invitation to participate in a survey or abandon a questionnaire before it is completed. Given today’s societal trend towards self-administration [2] the former is inevitable, irrespective of delivery mechanism. Conversely, non-response as a consequence of questionnaire abandonment can be relatively easily addressed. Unlike traditional questionnaires, the delivery mechanism for online-questionnaires makes estimation of questionnaire length and time required for completion difficult1, thus increasing the likelihood of abandonment. By incorporating a range of features into the design of an online questionnaire, it is possible to facilitate such estimation – and indeed, to provide respondents with context sensitive assistance during the response process – and thereby reduce abandonment while eliciting feelings of accomplishment [6]. For online-questionnaires, sampling error (“the result of attempting to survey only some, and not all, of the units in the survey population” [2, pg. 9]) can arise when all but a small portion of the anticipated respondent set is alienated (and so fails to respond) as a result of, for example, disregard for varying connection speeds, bandwidth limitations, browser configurations, monitors, hardware, and user requirements during the questionnaire design process. Similarly, measurement errors (“the result of poor question wording or questions being presented in such a way that inaccurate or uninterpretable answers are obtained” [2, pg. 11]) will lead to respondents becoming confused and frustrated. Sampling, measurement, and non-response errors are likely to occur when an online-questionnaire is poorly designed. Individuals will answer questions incorrectly, abandon questionnaires, and may ultimately refuse to participate in future surveys; thus, the benefit of online questionnaire delivery will not be fully realized. To prevent errors of this kind2, and their consequences, it is extremely important that practical, comprehensive guidelines exist for the design of online questionnaires. Many design guidelines exist for paper-based questionnaire design (e.g. [7-14]); the same is not true for the design of online questionnaires [2, 15, 16]. The research presented in this paper is a first attempt to address this discrepancy. Section 2 describes the derivation of a comprehensive set of guidelines for the design of online-questionnaires and briefly (given space restrictions) outlines the essence of the guidelines themselves. Although online-questionnaires reduce traditional delivery costs (e.g. paper, mail out, and data entry), set up costs can be high given the need to either adopt and acquire training in questionnaire development software or secure the services of a web developer. Neither approach, however, guarantees a good questionnaire (often because the person designing the questionnaire lacks relevant knowledge in questionnaire design). Drawing on existing software evaluation techniques [17, 18], we assessed the extent to which current questionnaire development applications support our guidelines; Section 3 describes the framework used for the evaluation, and Section 4 discusses our findings. Finally, Section 5 concludes with a discussion of further work.

Correlations between garden design plant applications and climate change

Possible effects of climate change means great challenges to landscape design professionals in Hungary. Our climate will shift towards the Mediterranean and we have to prepare for this with among others, choosing correctly the plants to be planted. Teaching garden design dendrology has not recognized yet the necessity and urgency of this matter. Quick measures are required due to the long life-time and slow development of woody taxons. This paper presents the double relationship between landscape design and climate change emphasizing the outdoor architectural methods of adjustment. Such techniques recognized abroad are presented like precipitation drainage by vegetation and extensive green roof. Finally the effects of climate change on ornamental plants application are presented together with the associated project started at the Corvinus University of Budapest in 2010.

Making the grade : the perceived impact on Florida's A+ plan on the learning environment of one elementary school

The purpose of this study was to ascertain the perceptions of educators at one elementary school regarding the changes in the teaching and learning environment and their related effects following the implementation of Florida's A+ high-stakes accountability system. This study also assessed whether these changes were identified by participants as meaningful and enduring, in terms of the definition by Lieberman and Miller (1999). Twenty-one educators, including 17 teachers and four administrators, at Blue Ribbon Elementary school were interviewed. Data were inductively coded and categorized into four major themes: (a) teaching and learning environment consistency, (b) changes in the teaching and learning environment since the implementation of A+, (c) effects of the changes, and (d) significant and enduring change. Findings fell into three categories (a) identified changes since A+ implementation, (b) effects of changes, and (c) what participants believed was significant and long term change, which included those characteristics of the school that had been identified as consistent in the teaching and learning environment. Statements of the participants explained their perceptions about what instructional decisions where made in response to the A+ Plan including the modification of curriculum, the addition or omission of subject matter taught, and the positive or negative impact these decisions had on the teaching and learning environment. It was found that study participants felt all changes and their effects were a direct result of the A+ Plan and viewed many of the changes as being neither significant nor long term Analysis of the educators' perceptions of the changes they experienced revealed the overall feeling that the changes were not indicative of what was necessary to make a school successful. For the participants, the changes lacked the characteristics that they had described as vital in what constituted success. This led to the conclusion that, by Lieberman and Miller's definition, the majority of changes and effects that were implemented at the school as a result of the mandated A+ Plan, were not meaningful and enduring for effective school reform.

Phonics Instruction using Pseudowords for Success in Phonetic Decoding

This study examined a Pseudoword Phonics Curriculum to determine if this form of instruction would increase students’ decoding skills compared to typical real-word phonics instruction. In typical phonics instruction, children learn to decode familiar words which allow them to draw on their prior knowledge of how to pronounce the word and may detract from learning decoding skills. By using pseudowords during phonics instruction, students may learn more decoding skills because they are unfamiliar with the “words” and therefore cannot draw on memory for how to pronounce the word. It was hypothesized that students who learn phonics with pseudowords will learn more decoding skills and perform higher on a real-word assessment compared to students who learn phonics with real words. Students from two kindergarten classes participated in this study. An author-created word decoding assessment was used to determine the students’ ability to decode words. The study was broken into three phases, each lasting one month. During Phase 1, both groups received phonics instruction using real words, which allowed for the exploration of baseline student growth trajectories and potential teacher effects. During Phase 2, the experimental group received pseudoword phonics instruction while the control group continued real-word phonics instruction. During Phase 3, both groups were taught with real-word phonics instruction. Students were assessed on their decoding skills before and after each phase. Results from multiple regression and multi-level model analyses revealed a greater increase in decoding skills during the second and third phases of the study for students who received the pseudoword phonics instruction compared to students who received the real-word phonics instruction. This suggests that pseudoword phonics instruction improves decoding skills quicker than real-word phonics instruction. This also suggests that teaching decoding with pseudowords for one month can continue to improve decoding skills when children return to real-word phonics instruction. Teacher feedback suggests that confidence with reading increased for students who learned with pseudowords because they were less intimidated by the approach and viewed pseudoword phonics as a game that involved reading “silly” words. Implications of these results, limitations of this study, and areas for future research are discussed.

The impact of standards-based practices in mathematics on the achievement of low-performing students

This study examined standards-based mathematics reform initiatives to determine if they would improve student achievement on the part of low-performing students. New curricula, the Carnegie Learning Cognitive Tutor®, were provided for algebra and geometry students. The new instructional strategy relied on both the teacher-led instruction and the use of computers to differentiate instruction for individual students. Mathematics teachers received ongoing professional development to help them implement the new curricula. In addition, teachers were provided with ongoing support to assist them with the transformation of the learning environments for students using standards-based practices. This quasi-experimental (nonrandomized) study involved teachers in two matched urban high schools. Analyses (ANCOVAs) revealed that the experimental group with an appropriately implemented program had significantly higher learning gains than the comparison group as determined by the students' 2007 mathematics Developmental Scale Score (DSS). In addition, the experimental group's adjusted mean for the second interim mathematics assessment was significantly higher than the comparison group's mean. The findings support the idea that if the traditional curriculum is replaced with standards-based curriculum, and the curriculum is implemented as intended, low-performing students may make significant learning gains. With respect to the teaching practices as observed with the Classroom Observation Protocol (COP), t-tests were conducted on four constructs. The results for both the algebra and geometry teachers on the constructs were not significant. The COP indicated that teachers in both the experimental and comparison groups used traditional instruction strategies in their classrooms. The analyses of covariance (ANCOVA) on the use of technology revealed no significant main effects for computer use.

Design, modelling, simulation and integration of cyber physical systems: methods and applications

The main drivers for the development and evolution of Cyber Physical Systems (CPS) are the reduction of development costs and time along with the enhancement of the designed products. The aim of this survey paper is to provide an overview of different types of system and the associated transition process from mechatronics to CPS and cloud-based (IoT) systems. It will further consider the requirement that methodologies for CPS-design should be part of a multi-disciplinary development process within which designers should focus not only on the separate physical and computational components, but also on their integration and interaction. Challenges related to CPS-design are therefore considered in the paper from the perspectives of the physical processes, computation and integration respectively. Illustrative case studies are selected from different system levels starting with the description of the overlaying concept of Cyber Physical Production Systems (CPPSs). The analysis and evaluation of the specific properties of a sub-system using a condition monitoring system, important for the maintenance purposes, is then given for a wind turbine.

Heritage Language Maintenance and Biliteracy Development for Immigrants' Children: A Study of Chinese Immigrants' Family Language Policy and Biliteracy Practices

This study reports on research that examines the family language policy (FLP) and biliteracy practices of middle-class Chinese immigrant families in a metropolitan area in the southwest of the U.S. by exploring language practices pattern among family members, language and literacy environment at home, parents’ language management, parents’ language attitudes and ideologies, and biliteracy practices. In this study, I employed mixed methods, including survey and interviews, to investigate Chinese immigrant parents’ FLP, biliteracy practices, their life stories, and their experience of raising and nurturing children in an English-dominant society. Survey questionnaires were distributed to 55 Chinese immigrant parents and interviews were conducted with five families, including mothers and children. One finding from this study is that the language practices pattern at home shows the trend of language shift among the Chinese immigrants’ children. Children prefer speaking English with parents, siblings, and peers, and home literacy environment for children manifests an English-dominant trend. Chinese immigrant parents’ language attitudes and ideologies are largely influenced by English-only ideology. The priority for learning English surpasses the importance of Chinese learning, which is demonstrated by the English-dominant home literacy practices and an English-dominant language policy. Parents invest more in English literacy activities and materials for children, and very few parents implement Chinese-only policy for their children. A second finding from this study is that a multitude of factors from different sources shape and influence Chinese immigrants’ FLP and biliteracy practices. The factors consist of family-related factors, social factors, linguistic factors, and individual factors. A third finding from this study is that a wide variety of strategies are adopted by Chinese immigrant families, which have raised quite balanced bilingual children, to help children maintain Chinese heritage language (HL) and develop both English and Chinese literacy. The close examination and comparison of different families with English monolingual children, with children who have limited knowledge of HL, and with quite balanced bilingual children, this study discovers that immigrant parents, especially mothers, play a fundamental and irreplaceable role in their children’s HL maintenance and biliteracy development and it recommends to immigrant parents in how to implement the findings of this study to nurture their children to become bilingual and biliterate. Due to the limited number and restricted area and group of participant sampling, the results of this study may not be generalized to other groups in different contexts.

Scenic Design of James and the Giant Peach

The purpose of this thesis is to provide research, supporting paperwork, production photographs and other materials that document the scenic design process for James and the Giant Peach at Adventure Theatre MTC. This thesis contains the following: concept statement, scenic research images collected to express location, and the emotional/ intellectual/ psychological landscapes for the production, preliminary sketches, photographs of the ¼” scale model, drafting plates and supporting paint elevations to communicate the design, prop list and accompanying research, archival production photographs to document the completed design, and finally periodical reviews of the show.

Combination of Evolutionary Algorithms with Experimental Design, Traditional Optimization and Machine Learning

Evolutionary algorithms alone cannot solve optimization problems very efficiently since there are many random (not very rational) decisions in these algorithms. Combination of evolutionary algorithms and other techniques have been proven to be an efficient optimization methodology. In this talk, I will explain the basic ideas of our three algorithms along this line (1): Orthogonal genetic algorithm which treats crossover/mutation as an experimental design problem, (2) Multiobjective evolutionary algorithm based on decomposition (MOEA/D) which uses decomposition techniques from traditional mathematical programming in multiobjective optimization evolutionary algorithm, and (3) Regular model based multiobjective estimation of distribution algorithms (RM-MEDA) which uses the regular property and machine learning methods for improving multiobjective evolutionary algorithms.

Designing Happiness: Architecture and urban design for joy and well-being

Scientific studies exploring the environmental and experiential elements that help boost human happiness have become a significant and expanding body of work. Some urban designers, architects and planners are looking to apply this knowledge through policy decisions and design, but there is a great deal of room for further study and exploration. This paper looks at definitions of happiness and happiness measurements used in research. The paper goes on to introduce six environmental factors identified in a literature review that have design implications relating to happiness: Nature, Light, Surprise, Access, Identity, and Sociality. Architectural precedents are examined and design strategies are proposed for each factor, which are then applied to a test case site and building in Baltimore, Maryland. It is anticipated that these factors and strategies will be useful to architects, urban designers and planners as they endeavor to design positive user experiences and set city shaping policy.

Investigation of Escape and Negative Student Behaviors Related to Florida State High Stakes Test Preparation in Miami-Dade County Public High Schools, having been approved in respect to style and intellectual content, is referred to you for judgment

The purpose of this study was to determine whether there was a relationship between pressure to perform on state mandated, high-stakes tests and the rate of student escape behavior defined as the number of school suspensions and absences. The state assigned grade of a school was used as a surrogate measure of pressure with the assumption that pressure increased as the school grade decreased. Student attendance and suspension data were gathered from all 33 of the regular public high schools in Miami-Dade County Public Schools. The research questions were: Is the number of suspensions highest in the third quarter, when most FCAT preparation takes place for each of the 3 school years 2007-08 through 2009-10? How accurately does the high school’s grade predict the number of suspensions and number of absences during each of the 4 school years 2005-06 through 2008-09? The research questions were answered using repeated measures analysis of variance for research question #1 and non-linear multiple regression for research question #2. No significant difference could be found between the numbers of suspensions in each of the grading periods nor was there a relationship between the number of suspensions and school grade. A statistically significant relationship was found between student attendance and school grade. When plotted, this relationship was found to be quadratic in nature and formed a loose inverted U for each of the four years during which data were collected. This indicated that students in very high and very low performing schools had low levels of absences while those in the midlevel of the distribution of school performance (C schools) had the greatest rates of absence. Identifying a relationship between the pressures associated with high stakes testing and student escape behavior suggests that it might be useful for building administrators to reevaluate test preparation activities and procedures being used in their building and to include anxiety reducing strategies. As a relationship was found, it sets the foundation for future studies to identify whether testing related activities are impacting some students emotionally and are causing unintended consequences of testing mandates.

Life Design Counseling process and outcome: A case study with an adolescent

This article aims to explore the relationship between clients´ narrative transformation and the promotion of vocational decidedness and career maturity in a mid-adolescent case of Life Design Counseling (LDC). To assess LDC outcomes the Vocational Certainty Scale and the Career Maturity Inventory – Form C were used before and after the intervention. To intensively analyze the process of LDC change two measures of narrative change were used: the Innovative Moments Coding System (IMCS), as a measure of innovation emergence, and the Return to the Problem Coding System (RPCS), as a measure of ambivalence towards change. The results show that the three LDC sessions produced a significant change in vocational certainty but not in career maturity. Findings confirm that the process of change, according to the IMCS, is similar to the one observed in previous studies with adults. Implications for future research and practice are discussed.