The presented thesis is dedicated to particle identification in COMPASS RICH-1 experiment which is located in European Organization for Nuclear Research (CERN). For particle identification Cherenkov radiation is used, which is described in the thesis together with detection principles. Current aim in detector upgrade is replacing a part of Multi Wire Proportional Chambers (MWPC) in peripheral regions of the detection surface by a suitable photon detector so as to guarantee one-photon detection. For this purpose the hybrid photon detector based on THGEM and MicroMegas technologies was developed. The hybrid detector test results are presented in the thesis.

Thesis (MSD) --Boston University, Henry M. Goldman School of Dental Medicine, 2009 (Department of Periodontology and Oral Biology). / Includes bibliographic references: leaves 52-59. / Platelet-rich plasma (PRP) is a concentrated gel of platelets that contains several growth factors. Growth factors have been recognized as the part of PRP that play role in regeneration of the bone. It is not clear how these growth factors in PRP affect the bone regeneration. Resolvin El (RvEl; 5S,12R,18R-trihydroxyeicosapentaenoic acid) is an pro-resolving lipid mediator derived from omega-3 fatty acid eicosapentaenoic acid and shown to have potent effects on the resolution of inflammation. The purpose of this study was to analyze the action of PRP and RVEl on the proliferation and behavior of osteoblasts and osteoclasts in vitro. PRP was prepared from 14 healthy donors. Osteoblast cultures were from a cell line (Saos2) of osteosarcoma cells. Osteoclasts were differentiated from primary human peripheral blood monocytes. Osteoclastic morphology was studied and activity was analyzed via resorption on dentin discs using SEM. PRP and RVE 1 were added at different doses and time-points. Osteoblast function was analyzed by osteocalcin expression and release. Osteoclast activity was assessed by resorption and cathepsin K expression. PRP and RvEl comparably increased the osteoblastic activity and suppressed the osteoclast differentiation and function. These results suggest that multiple tools are available to reverse the inflammation and restore the lost bone architecture as a result of periodontal disease.

Silicon based materials, including silicon oxides and silicon oxynitrides, have found use in a number of areas in photonics including waveguides, antireflection and highly reflective coatings for laser facets, and detectors. For effective use of these materials in photonics it is necessary to characterize their optical properties as a function of their composition and structure. Since these characteristics are often dependent on the method used to deposit the films it is necessary to also determine the effect of deposition type and conditions on the film's properties. Recently, silicon based materials have been seen to display luminescence due to quantum confinement effects when nanocrystals are formed. This opens up the possibility of a silicon based emitter, something that has not had previous success due to the indirect bandgap of bulk silicon. The development of a silicon based emitter in turn would open up the possibility for monolithically integrated photonic circuits that could take advantage of CMOS processing technology. This thesis presents the results of research into the characterization of amorphous silicon and silicon oxide thin films deposited by electron cyclotron resonance plasma enhanced chemical vapor deposition. Optical properties of the films have been determined through the use of ellipsometry and correlated with the results from compositional analysis, done using Rutherford backscattering and elastic recoil detection, and bonding structure analysis, done using Fourier transform infrared spectroscopy. Nanocrystals were formed within the films by subjecting them to post-deposition thermal annealing, which induces a phase separation in silicon rich silicon oxide films. The effects of different annealing conditions on composition, structure and optical properties have also been analyzed. Finally, photoluminescence experiments were conducted on the films and correlated with the results from other characterization techniques. / Thesis / Master of Applied Science (MASc)

The inherent design freedom of protein engineering and recombinant protein production enables specific tailoring of protein structure, function, and properties. Two areas of research where protein engineering has allowed for many advances in biomedical materials include the design of novel protein scaffolds for molecular recognition, as well as the use of recombinant proteins for production of next generation biomaterials. The main focus of my dissertation was to develop new biomedical materials using protein engineering. Chapters three and four discuss the engineering of repeat proteins as bio-recognition modules for biomedical sensing and imaging. Chapter three provides an overview of the most recent advances in engineering of repeat proteins in the aforementioned field. Chapter four discusses my contribution to this field. We have designed a de novo repeat protein scaffold based on the consensus sequence of the leucine rich repeat (LRR) domain of the NOD family of cytoplasmic innate immune system receptors. Innate immunity receptors have been described as pattern recognition receptors in that they recognize "global features" of a family of pathogens versus one specific antigen. In mammals, two main protein families of such receptors are: extracellular Toll-like receptors (TLRs) and cytoplasmic Nucletide-binding domain- and Leucine-rich Repeat-containing proteins (NLRs). NLRs are defined by their tripartite domain architecture that contains a C-terminal LRR (Leucine Rich Repeat) domain, the nucleotide-binding oligomerization (NACHT) domain, and the N-terminal effector domain. It is proposed that pathogen sensing in NLRs occurs through ligand binding by the LRR domain. Thus, we hypothesized that LRRs would be suitable for the design of alternative binding scaffolds for use in molecular recognition. The NOD protein family plays a very important role in innate immunity, and consequently serves as a promising scaffold for design of novel recognition motifs. However, engineering of de novo proteins based on the NOD family LRR domain has proven challenging due to problems arising from protein solubility and stability. Consensus sequence design is a protein design tool used to create novel proteins that capture sequence-structure relationships and interactions present in nature in order to create a stable protein scaffold. We implement a consensus sequence design approach to develop proteins based on the LRR domain of NLRs. Using a multiple sequence alignment we analyzed all individual LRRs found in mammalian NLRs. This design resulted in a consensus sequence protein containing two internal repeats and separate N- and C- capping repeats named CLRR2. Using biophysical characterization methods of size exclusion chromatography, circular dichroism, and fluorescence, CLRR2 was found to be a stable, monomeric, and cysteine free scaffold. Additionally, CLRR2, without any affinity maturation, displayed micromolar binding affinity for muramyl dipeptide (MDP), a bacterial cell wall fragment. To our knowledge, this is the first report of direct interaction of a NOD LRR with a physiologically relevant ligand. Furthermore, CLRR2 demonstrated selective recognition to the biologically active stereoisomer of MDP. Results of this study indicate that LRRs are indeed a useful scaffold for development of specific and selective proteins for molecular recognition, creating much potential for future engineering of alternative protein scaffolds for biomedical applications. My second research interest focused on the development of proteins for novel biomaterials. In the past two decades, keratin biomaterials have shown impressive results as scaffolds for tissue engineering, wound healing, and nerve regeneration. In addition to its intrinsic biocompatibility, keratin interacts with specific cell receptors eliciting beneficial biochemical cues, as well as participates in important regulatory functions such as cell migration and proliferation and protein signalling. The aforementioned properties along with keratins' inherent capacity for self-assembly poise it as a promising scaffold for regenerative medicine and tissue engineering applications. However, due to the extraction process used to obtain natural keratin proteins from natural sources, protein damage and formation of by-products that alter network self-assembly and bioactivity often occur as a result of the extensive processing conditions required. Furthermore, natural keratins require exogenous chemistry in order to modify their properties, which greatly limits sequence tunability. Recombinant keratin proteins have the potential to overcome the limitations associated with the use of natural keratins while also maintaining their desired structural and chemical characteristics. Thus, we have used recombinant DNA technology for the production of human hair keratins, keratin 31 (K31) and keratin 81 (K81). The production of recombinant human hair keratins resulted in isolated proteins of the correct sequence and molecular weight determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and mass spectrometry. Proteins with no unwanted sequence truncations, deletions, or mutations indicate recombinant DNA technology can be used to reliably generate full length keratin proteins. This allows for consistent starting materials with no observable impurities or undesired by-products, which combats a major challenge associated with natural keratins. Additionally, recombinant keratins must maintain the intrinsic propensity for self-assembly found in natural keratins. To test the propensity for self-assembly, we implemented size exclusion chromatography (SEC), dynamic light scattering (DLS), and transmission electron microscopy (TEM) to characterize K31, K81, and an equimolar mixture of K31 and K81. The results of the recombinant protein characterization reveal novel homo-polymerization of K31 and K81, not previously reported, and formation of characteristic keratin fibers for the K31 and K81 mixture. Therefore, recombinant K31 and K81 retain the intrinsic biological activity (i.e. self-assembly) of natural keratin proteins. We have also conducted a comparative study of recombinant and extracted heteropolymer K31/K81. Through solution characterization and TEM analysis it was found that use of the recombinant heteropolymer allows for increased purity of starting material while also maintaining self-assembly properties necessary for functional use in biomaterials design. However, under the processing condition implemented, extracted keratins demonstrated increased efficiency of assembly. Through each study we conclude that recombinant keratin proteins provide a promising solution to overcome the challenges associated with natural protein materials and present an exceptional design platform for generation of new biomaterials for regenerative medicine and tissue engineering. / Ph. D.

This thesis serves as an introduction to use of the NetBeans Platform for development of desktop applications. Its goal is to present the platform both on a compact example application and on a real-world scenario application. Secondary goal is to provide a summary of materials for further learning. The description of the particular pieces of the platform is interlaid with the step by step tutorial, which invites to further testing of the platform's features. Some principles of the platform are then presented in a source code from a real-world application. This application is a tool for authors and should provide an evidence of both academic and literary texts, notes and named entities (like characters, locations, terms etc.). The analysis of this tool and a description in a more detail is presented in the appendix. An organized summary of shortly commented links to other sources is there too. The main benefits of this thesis are: the demonstration of the NetBeans Platform on particular examples in context, and the summary of online materials for further reading.

Orientador: Maria Luiza Silveira Mello / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-20T03:24:19Z (GMT). No. of bitstreams: 1 Alvarenga_EleniceMonte_M.pdf: 2829254 bytes, checksum: 964b17aaf1c7f50d4471a7be569aaa6d (MD5) Previous issue date: 2011 / Resumo: A cromatina pode existir em núcleos interfásicos em dois estados distintos: como eucromatina e como heterocromatina, podendo ser esta constitutiva ou facultativa. Em células somáticas do final da fase ninfal dos hemípteros reduviídeos Triatoma infestans e Panstrongylus megistus há núcleos grandes, poliploides, nos quais a heterocromatina apresenta-se como corpos conspícuos (cromocentros), daí tais células apresentarem-se como um bom modelo para investigação de características morfológicas e funcionais da cromatina. Em estudos sobre a constituição cromatínica, a composição em bases do DNA é algo muito explorado, dado o conteúdo informativo dos achados. Já quando se objetiva a investigação da funcionalidade da cromatina, mais recentemente, tem-se feito uso da abordagem epigenética. Neste trabalho buscou-se investigar a composição em bases do DNA destas células, associando-a aos domínios cromatínicos aí existentes e também à presença das NORs. Por meio de colorações fluorescentes com Cromomicina A3 (CMA3)/Distamicina e 4',6-diamidino-2-fenilindol (DAPI)/Actinomicina D concluiuse que o DNA dos cromocentros de T. infestans e P. megistus são ricos em sequências AT e pobres em GC. Isto foi ainda confirmado por imunodetecção de 5-metilcitosina, que ocorreu somente na eucromatina, e tratamento de ninfas de T. infestans com 5-aza-2'-deoxicitidina (agente demetilante), seguido da análise dos fenótipos nucleares e análise de imagem, em que se observou expansão somente da área eucromática. Com o método de AgNOR evidenciou-se que a região rica em bases GC ao redor do cromocentro coincide com um acúmulo de proteínas argirofílicas, o que sugere associação com NORs. A presença de modificações epigenéticas nas caudas das histonas na cromatina destes insetos foi investigada por meio do uso de anticorpos contra marcadores epigenéticos específicos, permitindo identificar a participação diferencial dos mesmos na composição e na estrutura dos territórios heterocromáticos. Assim, observou-se hipoacetilação e hipermetilação de histonas na região do corpo heterocromático, o que indicaria uma possível ação da modificação de histonas na manutenção da estrutura heterocromática nas células somáticas de ambas as espécies de reduviídeos. Por meio da avaliação da ação de drogas inibidoras de deacetilases de histonas sobre a cromatina dos insetos percebeu-se que, quando ninfas de T. infestans e P. megistus foram tratadas com as drogas, houve aumento na frequência de necroses e, no caso específico do tratamento com tricostatina A (TSA) e butirato de sódio (NaBt), ocorreu descompactação da heterocromatina. Sugere-se que o tratamento com TSA e NaBt afete o processo de deacetilação de histonas, o qual seria, então, um fator importante na estruturação dos cromocentros. A observação da ocorrência de mudas e da sobrevivência de ninfas de T. infestans, realizada a fim de se avaliar a ação do ácido valproico (VPA) sobre o desenvolvimento dos insetos, mostrou que a droga, assim como a injeção de solução salina, reduziu seu período de sobrevivência, além de afetar a ocorrência de mudas / Abstract: Chromatin in interphase cell nuclei can be present in two distinct states: euchromatin and heterochromatin, which may be constitutive or facultative. In somatic cells at the end of the nymphal stage of Triatoma infestans and Panstrongylus megistus there are large nuclei, in which heterochromatin is presented as conspicuous bodies (chromocenters). These cells are an appropriate model for investigation of morphological and functional characteristics of the chromatin. In studies about chromatin constitution, the base DNA composition is explored due to the informational content of the findings. If the objective is to investigate the chromatin functionality, recently has been used the epigenetic approach. In the current study, the aim was to investigate the DNA base composition in these cells, associating with the chromatin domains therein and also the presence of NORs. Through fluorescent stains with Chromomycin A3 (CMA3)/Distamycin and 4',6-diamidino-2-fenilindole (DAPI)/Actinomycin D was found that the chromocenters DNA of T. infestans and P. megistus were AT-rich and GC-poor. This was also confirmed by immunodetection of 5-methylcytidine, which occurred only in the euchromatin, and by T. infestans nymphs treatment with 5-aza-2'- deoxycytidine (demethylating agent), followed by nuclear phenotypes analysis and image analysis, in which expansion was observed only in the euchromatic area. AgNOR test evidenced that the GC-rich region around the chromocenter coincides with an accumulation of argyrophilic proteins, suggesting association with NORs. Epigenetic modifications on histone tails in chromatin of these insects were investigated by using antibodies against specific epigenetic markers, in order to identify their differential participation in the composition and structure of these heterochromatic regions. It was observed hypoacetylation and hypermethylation in heterochromatic body area, suggesting a possible action of histones modification in the maintenance of the heterochromatic structure in somatic cells of both species of reduviids. Through evaluation of histones deacetylases inhibitors action on the chromatin, it was observed that when T. infestans and P. megistus nymphs were treated with these drugs there was an increase in the frequencies of necrosis, and in the specific case of Trichostatin A (TSA) and sodium butyrate (NaBt), occurred heterochromatin decondensation. It is suggested that treatments with TSA and NaBt could affect the histones deacetilation process, which would be an important factor in chromocenters structuring. Observations of the molting occurrence and survival of T. infestans nymphs, carried out in order to evaluate the action of valproic acid (VPA) on the development of insects, showed that this drug, as well as injection of saline, reduced the survival period, besides affecting the molting occurrence / Mestrado / Biologia Celular / Mestre em Biologia Celular e Estrutural

Thesis (MBA)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: This research study has sought to gain a deeper understanding of the manner in which the ultra-luxury travel sector interprets the concept of customer value and how they go about designing an offering of potential value to the customer. In turn, the study explored the manner in which the traveller extracts value from the offering. There is a definite element of ambiguity to the precise meaning of the word ‘luxury’. Researchers have traced its meaning and developed frameworks to define the type of value it offers to the consumer. Ubiquitous to the delivery of the luxury good or service, is the significance that an individual will attach to it. The consumption of luxury will always be an experience – in one form or another. With the passing of time and globalisation of companies, the luxury good has become ‘democratised’. This massification of luxury has resulted in layers of offering, appealing to various categories of consumer; from the connoisseur and consumer of rare, niche products, to the bling young professional sporting a designer bag. This has created greater debate around the definition of luxury. Further frameworks have been developed to identify the types of value within the luxury offering that would appeal to the different categories of luxury consumer. The process of value creation has evolved from a firm-centred model to one where the customer is central. The focus is on identifying customer needs, and then delivering them via the use of firm competencies and capabilities. The lens of the value creation process has widened to include means of identifying potential for firm value creation pre and post consumption of the offering. The luxury travel sector generates a significant spend. As an industry that is intensely customer-centric, it provides the perfect vehicle to examine how the industry seeks to understand the guest, and in turn, deliver value. A qualitative, multiple case study methodology was employed to examine four luxury properties in South Africa. Employees and independent consultants within the industry were interviewed. The results yielded a coherent message of passionate engagement with the product and the guest. The surprising element was the employee’s fierce commitment to the luxury asset and their pride in the owner. The element of sustainability featured strongly as, whilst every effort was made to give the guest a magical experience, this was not done at the expense of the underlying asset. There will always be an element of experience attached to the immersion in a luxury product or service and the luxury travel industry is no exception. Experiential travel has been a driving force within the industry for many years. As this evolves, the pressure to plumb greater depths of meaning and create significant shifts in ‘being’ has developed. The industry is ripe for creative collaborations and new partnerships in order to deliver life-long memories to the luxury traveller. Technology has been a disrupter within the luxury value chain and tour operators and travel agents find themselves competing with online travel agents. Those who survive are those who deliver highly personalised, creative and bespoke offerings. Social media is another area of potential innovation. The Millennial traveller will soon command the lion’s share of luxury spending. They are highly connected and desire authentic, fun experiences. Luxury accommodation has also suffered the ambiguity attached to the definition of luxury brands in general. The 5-star hotel which once commanded pride of place as the highest level of accommodation has been usurped by private luxury properties, boutique and destination hotels. The potential exists to apply the lenses of luxury value frameworks in order to provide greater clarity to the type of offering they deliver to the luxury traveller.

Web applications have come a long way both in terms of adoption to provide information and services and in terms of the technologies to develop them. With the emergence of richer and more advanced technologies such as Ajax, web applications have become more interactive, responsive and user friendly. These applications, often called Rich Internet Applications (RIAs) changed the traditional web applications in two primary ways: Dynamic manipulation of client side state and Asynchronous communication with the server. At the same time, such techniques also introduce new challenges. Among these challenges, an important one is the difficulty of automatically crawling these new applications. Crawling is not only important for indexing the contents but also critical to web application assessment such as testing for security vulnerabilities or accessibility. Traditional crawlers are no longer sufficient for these newer technologies and crawling in RIAs is either inexistent or far from perfect. There is a need for an efficient crawler for web applications developed using these new technologies. Further, as more and more enterprises use these new technologies to provide their services, the requirement for a better crawler becomes inevitable. This thesis studies the problems associated with crawling RIAs. Crawling RIAs is fundamentally more difficult than crawling traditional multi-page web applications. The thesis also presents an efficient RIA crawling strategy and compares it with existing methods.

Mobile Digital TV (MDTV), the hybrid of Digital Television (DTV) and mobile devices (such as mobile phones), has introduced a new way for people to watch DTV and has brought new opportunities for development in the DTV industry. Nowadays, the development of the next generation MDTV service has progressed in terms of both hardware layers and software, with interactive services/applications becoming one of the future MDTV service trends. However, current MDTV interactive services still lack in terms of attracting the consumers and the service creation and implementation process relies too much on commercial solutions, resulting in most parts of the process being proprietary. In addition, this has increased the technical demands for developers as well as has increased substantially the cost of producing and maintaining MDTV services. In light of the aforementioned situation, the Thesis has contributed to this field, by proposing an innovative MDTV service creation and consumption system based on XHTML and Java ME. On the head-end it introduces a semi-automatic creation mechanism to facilitate a less technical and more efficient interactive service creation process. This enables designers and creative individuals to be actively involved in the MDTV service creation process and to develop interactive-rich MDTV service. On the client-end it employs an open-source software environment as the interactive service MDTV consumption platform, rendering the MDTV service implementation process as less proprietary as possible. Furthermore, the Thesis offers a discussion on the different MDTV interactive application models currently used and based on the proposed software, a novel MDTV service presentation method is further introduced and adopted instead of the Rich Media and ECMAScript based methods. Finally, a series of qualitative testing procedures have been implemented with regards to conducting an essential evaluation on the operability of the proposed software system.

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