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Adipose tissue is a type of connective tissue whose purpose was once thought to be limited to fat storage but is now understood to be a key factor in the pathogenesis of different metabolic diseases, including obesity and type-II diabetes. Adipose tissue consists largely of adipocytes, cells responsible for fat and releasing energy in form of lipids. Different classes of fatty acids, such as saturated and unsaturated have different biological effects on adipocytes. Lipid droplets are the primary organelles in adipocytes that store these fatty acids in form of lipids, and the development of engineered adipose tissues would benefit from improved techniques for analysis of lipid droplet composition, distributions, and dynamics based as a function of fatty acid saturation. Conventional microscopic techniques, such as fluorescence microscopy, provides excellent selectivity of lipid-based structures inside adipose tissue cellular structures based on staining with compound dyes. However, fluorescence staining limits multiplex imaging, and requires time consuming steps in preparing the samples for imaging. Therefore, developing a label-free, high resolution imaging platform with sensitivity to lipid composition could enable analysis of structural and compositional differentiation of lipid droplets within adipocytes during differentiation could give valuable insights into the importance of lipid droplets role in metabolism. As an important step towards achieving this goal, a femtosecond based CARS microscopy imaging platform has been developed to perform in vitro, label-free, imaging of fatty acid composition within engineered adipose tissues. / Bioengineering

Master of Science / A quarter of diabetics suffer from diabetic foot ulcers due to chronic nerve damage, a leading complication found in many patients. Symptoms cause changes in the skin's environment, leading to frequent cuts in the skin. This often results in recurrent foot ulcers that are at high risk of infections. Currently, typical treatments take the form of antibiotics, which can lead to serious fungal or yeast infections. Candida albicans is one of the leading non-bacterial pathogens causing infections in diabetic foot ulcers, and if left untreated, these infections can progress into candidiasis. Candidiasis causes inner tissue damage that often requires amputations or, in severe cases, enters the bloodstream, leading to death. Candida infections are typically treated with azoles like fluconazole. There is an unmet need to develop new strategies to detect and treat Candida infections. In this work, we developed hydrogel-based Engineered Living Materials (ELMs) as an antifungal wound dressing. Additionally, to increase the efficacy of engineered antifungal dressing, we develop a genetic drug releasing system in the presence of blue light. Thus, here we describe the fundamental work-frame for the development of smart wound dressing to treat various fungal and bacterial infections.

A contemporaneidade vem sendo marcada por mudanças em vários aspectos, em especial para o design. As últimas décadas inauguraram tecnologias digitais em que os recursos para captura, tratamento e reprodução de imagens alteram modos de desenvolvimento, produção e percepção estética. Este estudo refere-se ao design de superfície, especialidade do design, especificamente à estamparia têxtil digital. Tem como propósito investigar os recursos digitais voltados à estamparia têxtil como produtores de elementos visuais causadores de sentido e significado tendo em vista a relação: produção criativa, produto (aspectos subjetivos da estampa) e diferenciação. A pesquisa parte da abordagem qualitativa de caráter exploratório, por meio de referências bibliográficas, observação em estamparia digital selecionada na cidade de São Paulo e entrevistas a designers de superfície atuantes. Este estudo também propõe uma reflexão sobre alguns aspectos diferenciais e de subjetividade que as estampas permitem em produtos de moda em vestuário e decoração. Ao identificar e discutir possibilidades, potencialidades e limitações da estamparia digital nas áreas mencionadas, sugere-se contribuições para o design de superfície na criação de estamparia com fotografia a partir da demanda por diferenciação, em engineered print, tendo a peça impressa como suporte de arte e comunicação. / The contemporaneity has been marked by changes in various aspects, in particular for design. The latest decades inaugurated digital technologies in which the capabilities for capturing, processing and reproducing images changes modes of development, production and aesthetics perception. This study refers to the surface design, specialty of design, specifically the digital textile printing. It aims to investigate the digital resources intended for the textile printing as producers of visual elements that causes sense and meaning to the partnership: creative production, product (subjective aspects of print) and differentiation. This is a qualitative research of exploratory character approach, by means of bibliographical references, observation of selected digital prints in the city of São Paulo and interviews of the active surface designers. This study also proposes a reflection on some differential and subjective aspects that prints allows in fashion and decoration products. Identifying and discussing possibilities, potentials and limitations of digital printing in the areas mentioned, it suggests contributions to surface design in the creation of engineered prints from the differentiation demand, making the piece printed as art and communication support.

The 18 September 2000 disclosure that StarLink corn, a genetically engineered variety not approved for human consumption, had been detected in food was a seminal event in agricultural biotechnology. This thesis presents a comprehensive case study of the StarLink incident (part one), reviews the StarLink situation in terms of crisis management theory (part two) and develops crisis management theory using the StarLink incident as an example of a crisis (part three). Part one provides background information, then a meticulous day-by-day account of StarLink-related events. Part two presents a detailed overview of crisis management theory, then examines the StarLink situation in terms of pre-crisis (warning signals, preconditions for a crisis, crisis trigger), crisis (how Aventis, the biotechnology provider, managed the crisis and opinions concerning crisis handling) and post-crisis (lessons learned). Part three develops crisis management theory using the StarLink situation as an example of a crisis. It evaluates whether the StarLink incident possessed characteristics predicted for modern crises and suggests other factors which may become more prevalent and significant in future crises. The StarLink incident delivers certain practical lessons for managers, regulators and others and demonstrates a number of characteristics of modern crises.

A contemporaneidade vem sendo marcada por mudanças em vários aspectos, em especial para o design. As últimas décadas inauguraram tecnologias digitais em que os recursos para captura, tratamento e reprodução de imagens alteram modos de desenvolvimento, produção e percepção estética. Este estudo refere-se ao design de superfície, especialidade do design, especificamente à estamparia têxtil digital. Tem como propósito investigar os recursos digitais voltados à estamparia têxtil como produtores de elementos visuais causadores de sentido e significado tendo em vista a relação: produção criativa, produto (aspectos subjetivos da estampa) e diferenciação. A pesquisa parte da abordagem qualitativa de caráter exploratório, por meio de referências bibliográficas, observação em estamparia digital selecionada na cidade de São Paulo e entrevistas a designers de superfície atuantes. Este estudo também propõe uma reflexão sobre alguns aspectos diferenciais e de subjetividade que as estampas permitem em produtos de moda em vestuário e decoração. Ao identificar e discutir possibilidades, potencialidades e limitações da estamparia digital nas áreas mencionadas, sugere-se contribuições para o design de superfície na criação de estamparia com fotografia a partir da demanda por diferenciação, em engineered print, tendo a peça impressa como suporte de arte e comunicação. / The contemporaneity has been marked by changes in various aspects, in particular for design. The latest decades inaugurated digital technologies in which the capabilities for capturing, processing and reproducing images changes modes of development, production and aesthetics perception. This study refers to the surface design, specialty of design, specifically the digital textile printing. It aims to investigate the digital resources intended for the textile printing as producers of visual elements that causes sense and meaning to the partnership: creative production, product (subjective aspects of print) and differentiation. This is a qualitative research of exploratory character approach, by means of bibliographical references, observation of selected digital prints in the city of São Paulo and interviews of the active surface designers. This study also proposes a reflection on some differential and subjective aspects that prints allows in fashion and decoration products. Identifying and discussing possibilities, potentials and limitations of digital printing in the areas mentioned, it suggests contributions to surface design in the creation of engineered prints from the differentiation demand, making the piece printed as art and communication support.

Favorable properties of Balsa wood make it an interesting alternative in a number of applications including thermal insulation or as a lightweight core material in sandwich composites. Increasing use in construction necessitates a better understanding of its mechanical and failure properties. In the present work, mode I and mode II fracture toughness for different types of balsa wood and a sandwich structure (balsa as core and fiber glass as skin layer) are studied experimentally by using load-displacement diagrams and visually acquired crack growth data. / Graduation date: 2013

Thesis (MScEng (Process Engineering))--University of Stellenbosch, 2011. / Please refer to full text for abstracts

Several tools were developed to help researchers facilitate clinical translation of the use of engineered nucleases towards their disease gene of interest. Two major issues addressed were the inability to accurately predict nuclease off-target sites by user-friendly \textit{in silico} methods and the lack of a high-throughput, sensitive measurement of gene editing activity at endogenous loci. These objectives were accomplished by the completion of the following specific aims. An online search interface to allow exhaustive searching of a genome for potential nuclease off-target sites was implemented. Previously discovered off-target sites were collated and ranking algorithms developed that preferentially score validated off-target sites higher than other predictions. HEK-293T cells transfected with newly developed TALENs and ZFNs targeting the beta-globin gene were analyzed at the off-target sites predicted by the tool. Many samples of genomic DNA from cells treated with different ZFNs and TALENs were analyzed for off-target effects to generate a greatly expanded training set of bona fide off-target sites. Modifications to the off-target prediction algorithm parameters were evaluated for improvement through Precision-Recall analysis and several other metrics. An analysis pipeline was developed to process SMRT reads to simultaneously measure the rates of different DNA repair mechanisms by directly examining the DNA sequences. K562 cells were transfected with different types of nucleases and donor repair templates in order to optimize conditions for repairing the beta-globin gene. This work will have significant impact on future studies as the methods developed herein allow other laboratories to optimize nuclease-based therapies for single gene disorders.

The development of small-diameter artificial blood vessels that mimic the properties of natural blood vessels has proven to be a clinical challenge. While autologous vessels are the standard, they can be difficult to obtain and require invasive surgeries. Synthetic materials have been successful in large diameter applications, but they have been unsuccessful in small-caliber environments due to a number of factors including thrombus formation, intimal hyperplasia, and infection. Intimal hyperplasia, of particular interest in this study, involves the build up of smooth muscle cells (SMCs) in the intimal layer of the artery due to abnormal migration and proliferation. This work focuses on the development of a new polymer that has the potential to function as an intimal/medial component of a small-diameter blood vessel. Using recombinant elastin-like polypeptides (ELPs) developed by the Woodhouse laboratory, as well as chondroitin sulphate-specific binding sequences (CSBD1 and CSBD2) determined by the Panitch laboratory, a new elastin-like polypeptide-chondroitin sulphate binding domain (ELP-CSBD) block copolymer has been developed and characterized. The expression of the ELP1-CSBDs was accomplished using E. coli BL21 cells in a bioreactor or shaker flask systems. The polypeptides were purified using dialysis and ion exchange chromatography and expression and purity were characterized using mass spectrometry and amino acid analysis. Both ELP1-CSBDs were successfully expressed using these methods and ELP1-CSBD1 was produced to high purities. ELP1-CSBD1 was able to undergo coacervation in vitro, suggesting that ELP1-CSBD1 is able to self-assemble in a manner similar to native elastin. In the presence of the glycosaminoglycan chondroitin sulphate (CS), the temperature of coacervation of ELP1-CSBD1 is increased, the rate and extent of coacervation is decreased, and aggregates remain in solution even at higher temperatures. The influence of heparin was also explored as previous studies indicated that the CS binding domains were shown to also bind to heparin. Studies completed in the presence of heparin showed that there were no significant changes to the coacervation characteristics of ELP1-CSBD1. It is anticipated that when combined with CS, ELP1-CSBD1 will gel, forming a basis for an intimal/medial layer of a TEBV that will modulate SMC response and increase graft integrity. / Thesis (Master, Chemical Engineering) -- Queen's University, 2013-01-06 21:03:37.788