Dissertations / Theses on the topic 'It is likely that S'

Made available in DSpace on 2014-06-11T19:26:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-08-29Bitstream added on 2014-06-13T20:33:44Z : No. of bitstreams: 1 ondei_ls_me_sjrp.pdf: 1659909 bytes, checksum: 0234670a460d2993f23cb77243c91e66 (MD5)<br>As hemoglobinas (Hb) variantes, originadas em sua maioria por simples substituições de aminoácidos, resultam de mudanças na seqüência de nucleotídeos. Atualmente, o número de Hb anormais identificadas tem aumentado devido à melhoria nas metodologias de análises, porém muitos laboratórios de rotina não estão preparados para a correta identificação dos mutantes. A Hb S é uma variante de Hb bem caracterizada que apresenta prevalência variável nas diferentes regiões do Brasil. No entanto, há uma variedade de Hb que apresenta padrão de migração eletroforética semelhante ao da Hb S em pH alcalino as quais são denominadas “Hb S-like”, podendo ser erroneamente diagnosticadas e com suas freqüências subestimadas. Neste trabalho foram estabelecidos os valores referenciais de Hb por HPLC em portadores de Hb S e determinados os perfis eletroforético e cromatográfico das Hb “S-like” dentre as amostras enviadas ao LHGDH. Foram encontradas as Hb Hasharon, Hb D-Los Angeles, Hb Korle-Bu, Hb Lepore, Hb D-Iran, Hb tipo G, Hb Queens, Hb Montgomery e Hb Q-Índia. Também foram encontrados casos de associação entre dois mutantes de cadeia beta. As eletroforeses em pH alcalino e pH ácido foram utilizadas para o rastreamento inicial das Hb variantes e as eletroforeses de cadeias polipeptídicas em ambos pH foram realizadas para a identificação da cadeia globínica alterada. As análises cromatográficas permitiram o direcionamento dos prováveis mutantes, sendo também possível a quantificação precisa das frações de variantes. Desta forma, a associação de metodologias laboratoriais clássicas de diagnóstico é fundamental para o levantamento das suspeitas fenotípicas. Os perfis da Hb S e das Hb “S-like” estabelecidos neste estudo auxiliarão no diagnóstico dessas Hb variantes em serviços de saúde.<br>The variants hemoglobin (Hb) originated mainly by simple amino acid substitutions, result of nucleotides sequences changes. Currently, the number of abnormal hemoglobin identified has increased due to improvement in the analysis methodologies, but many routine laboratories are not prepared for the correct mutants identification. The Hb S is a variant Hb well characterized that shows variable prevalence in different regions of Brazil. However, there is a diversity of Hb that present electrophoretic migration on alkaline pH similar to Hb S, named Hb S-like, which can be incorrectly diagnosed and with their frequencies underestimated. At the present study the reference ranges for Hb S obtained by HPLC were established, and the electrophoretic and cromatographic profile for Hb S like were determined. The Hb Hasharon, Hb D-Los Angeles, Hb Korle-Bu, Hb Lepore, Hb D-Iran, Hb tipo G, Hb Queens, Hb Montgomery e Hb Q-Índia were found. Cases of association between two mutants of beta chain were also found. The electrophoresis on alkaline pH and acid pH were utilized to the initial screen of these variants Hb and the globin chains electrophoresis in both pH were performed to identify the mutant globin chain. The chromatographic analysis permitted the identification of the likely variant hemoglobin and also facilitated the exact quantification of variants. Therefore, the association of the classical laboratory methods of diagnostic is fundamental for the identification of variant Hb suspect. The Hb S and Hb S-like profile determined in this study will help in the diagnostic of these variants Hb in healthy service.

Orientador: Claudia Regina Bonini Domingos<br>Banca: Ivan de Lucena Angulo<br>Banca: Carlos Roberto Ceron<br>Resumo: As hemoglobinas (Hb) variantes, originadas em sua maioria por simples substituições de aminoácidos, resultam de mudanças na seqüência de nucleotídeos. Atualmente, o número de Hb anormais identificadas tem aumentado devido à melhoria nas metodologias de análises, porém muitos laboratórios de rotina não estão preparados para a correta identificação dos mutantes. A Hb S é uma variante de Hb bem caracterizada que apresenta prevalência variável nas diferentes regiões do Brasil. No entanto, há uma variedade de Hb que apresenta padrão de migração eletroforética semelhante ao da Hb S em pH alcalino as quais são denominadas "Hb S-like", podendo ser erroneamente diagnosticadas e com suas freqüências subestimadas. Neste trabalho foram estabelecidos os valores referenciais de Hb por HPLC em portadores de Hb S e determinados os perfis eletroforético e cromatográfico das Hb "S-like" dentre as amostras enviadas ao LHGDH. Foram encontradas as Hb Hasharon, Hb D-Los Angeles, Hb Korle-Bu, Hb Lepore, Hb D-Iran, Hb tipo G, Hb Queens, Hb Montgomery e Hb Q-Índia. Também foram encontrados casos de associação entre dois mutantes de cadeia beta. As eletroforeses em pH alcalino e pH ácido foram utilizadas para o rastreamento inicial das Hb variantes e as eletroforeses de cadeias polipeptídicas em ambos pH foram realizadas para a identificação da cadeia globínica alterada. As análises cromatográficas permitiram o direcionamento dos prováveis mutantes, sendo também possível a quantificação precisa das frações de variantes. Desta forma, a associação de metodologias laboratoriais clássicas de diagnóstico é fundamental para o levantamento das suspeitas fenotípicas. Os perfis da Hb S e das Hb "S-like" estabelecidos neste estudo auxiliarão no diagnóstico dessas Hb variantes em serviços de saúde.<br>Abstract: The variants hemoglobin (Hb) originated mainly by simple amino acid substitutions, result of nucleotides sequences changes. Currently, the number of abnormal hemoglobin identified has increased due to improvement in the analysis methodologies, but many routine laboratories are not prepared for the correct mutants identification. The Hb S is a variant Hb well characterized that shows variable prevalence in different regions of Brazil. However, there is a diversity of Hb that present electrophoretic migration on alkaline pH similar to Hb S, named Hb S-like, which can be incorrectly diagnosed and with their frequencies underestimated. At the present study the reference ranges for Hb S obtained by HPLC were established, and the electrophoretic and cromatographic profile for Hb S like were determined. The Hb Hasharon, Hb D-Los Angeles, Hb Korle-Bu, Hb Lepore, Hb D-Iran, Hb tipo G, Hb Queens, Hb Montgomery e Hb Q-Índia were found. Cases of association between two mutants of beta chain were also found. The electrophoresis on alkaline pH and acid pH were utilized to the initial screen of these variants Hb and the globin chains electrophoresis in both pH were performed to identify the mutant globin chain. The chromatographic analysis permitted the identification of the likely variant hemoglobin and also facilitated the exact quantification of variants. Therefore, the association of the classical laboratory methods of diagnostic is fundamental for the identification of variant Hb suspect. The Hb S and Hb S-like profile determined in this study will help in the diagnostic of these variants Hb in healthy service.<br>Mestre

Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2009.<br>Cataloged from PDF version of thesis.<br>Includes bibliographical references (p. 89-93).<br>In this work, I test the hypothesis that the 2-dimensional dependencies of a deterministic model can be correctly recovered via hypothesis-enumeration and Bayesian selection for a linear sequence, and what the degree of 'ignorance' or 'uncertainty' is that Bayesian selection can tolerate concerning the properties of the model and data. The experiment tests the data created by a number of rules of size 3 and compares the implied dependency map to the (correct) dependencies of the various generating rules, then extends it to a composition of 2 rules of total size 5. I found that 'causal' belief networks do not map directly to the dependencies of actual causal structures. For deterministic rules satisfying the condition of multiple involvement (two tails), the correct model is not likely to be retrieved without augmenting the model selection with a prior high enough to suggest that the desired dependency model is already known - simply restricting the class of models to trees, and placing other restrictions (such as ordering) is not sufficient. Second, the identified-model to correct-model map is not 1 to 1 - in the rule cases where the correct model is identified, the identified model could just as easily have been produced by a different rule. Third, I discovered that uncertainty concerning identification of observations directly resulted in the loss of existing information and made model selection the product of pure chance (such as the last observation). How to read and identify observations had to be agreed upon a-priori by both the rule and the learner to have any consistency in model identification.<br>(cont.) Finally, I discovered that it is not the rule-observations that discriminate between models, but rather the noise, or uncaptured observations that govern the identified model. In analysis, I found that in enumeration of hypotheses (as dependency graphs) the differentiating space is very small. With representations of conditional independence, the equivalent factorizations of the graphs make the differentiating space even smaller. As Bayesian model identification relies on convergence to the differentiating space, if those spaces are diminishing in size (if the model size is allowed to grow) relative to the observation sequence, then maximizing the likelihood of a particular hypothesis may fail to converge on the correct one. Overall I found that if a learning mechanism either does not know how to read observations or know the dependencies he is looking for a-priori, then it is not likely to identify them probabilistically. Finally, I also confirmed existing results - that model selection always prefers increasingly connected models over independent models was confirmed, as was the knowledge that several conditional-independence graphs have equivalent factorizations. Finally Shannon's Asymptotic Equipartition Property was confirmed to apply both for novel observations and for an increasing model/parameter space size. These results are applicable to a number of domains: natural language processing and language induction by statistical means, bioinformatics and statistical identification and merging of ontologies, and induction of real-world causal dependencies.<br>by Beracah Yankama.<br>S.M.

Post-translational protein modifications (PTM) vastly increase the complexity and functional diversity of the proteome, to precisely regulate crucial cellular processes. The plant immune system is composed of complex signalling networks that are influenced by various PTMs. Activation of plant immunity is associated with a rapid burst of nitric oxide (NO), which can covalently modify cysteine thiols within target proteins by a process termed S-nitrosylation to form S-nitrosothiols (SNOs), constituting a redox-based PTM. Another key PTM involved in plant immunity is SUMOylation, an essential mechanism involving the conjugation of the small ubiquitin-like modifier (SUMO) peptide to lysine residues within target proteins. Although the targets and mechanisms of S-nitrosylation and SUMOylation are becoming evident, how these key PTMs are themselves regulated remains obscure. Work presented in this thesis reveals that during plant immune signalling, the sole Arabidopsis thaliana SUMO conjugating enzyme, SUMO CONJUGATING ENZYME 1 (SCE1), is S-nitrosylated at a highly conserved, but previously uncharacterized cysteine. S-nitrosylation of SCE1 was shown to inhibit its SUMO conjugating activity in vitro and mutational analysis revealed that the site of this modification, Cys139, is not required for enzyme activity but rather constitutes a redox-sensitive inhibitory switch. Generation and characterization of transgenic Arabidopsis plants overexpressing both wild-type and mutant forms of SCE1 revealed that Cys139 is required for efficient immunity against bacterial pathogens. Furthermore, after immune activation, S-nitrosylation of this residue inhibits global SUMOylation of proteins. These results provide evidence of a novel means of crosstalk between S-nitrosylation and SUMOylation in the context of plant immunity. The abundant cellular antioxidant, glutathione (GSH), is S-nitrosylated to form S-nitrosoglutathione (GSNO), which is thought to constitute a stable reservoir of NO bioactivity. In Arabidopsis, GSNO levels are controlled by the enzyme S-NITROSOGLUTATHIONE REDUCTASE 1 (GSNOR1), which indirectly influences the levels of protein SNOs. In this study, transgenic plants overexpressing FLAG-epitope tagged GSNOR1 were generated in various mutant backgrounds, including nitric oxide overproducer 1 (nox1), to further investigate the roles of GSNOR1 and NO in plant immunity. It was shown that ectopic GSNOR1 expression completely recovers developmental and disease susceptibility phenotypes of gsnor1, but not nox1 mutant plants, highlighting in vivo differences between accumulation of GSNO and free NO. Surprisingly, elevated NO levels in nox1 plants promote S-nitrosylation of GSNOR1, inhibiting its enzymatic activity. This suggests a previously unreported means by which NO might regulate its own bioavailability. Further work in this study revealed that recombinant GSNOR1 can be SUMOylated in vitro, which appeared to increase its enzymatic activity. Several potential SUMO modification sites were identified within GSNOR1 and mutational analysis revealed that at least one of these, Lys191, is SUMOylated. Co-immunoprecipitation experiments revealed that transgenic GSNOR1 might be SUMOylated in vivo, although the site(s) and biological function of SUMOylation were not identified. Nonetheless, these results reveal another possible layer of interplay between S-nitrosylation and SUMOylation.

This thesis deals with the image object detection using Haar--like features and AdaBoost algorithm. The text describes methods how to train and test an object detector. The main contributon of this thesis consists in creation image object detector in Java programming language. Created algorithms were integrated as plugin into the RapidMiner tool, which is widely used and known worldwide as tool for data mining. The thesis contains the instructions for created operators and few exaples for executing in RapidMiner tool. The functionality of image object detector was demonstrated on selected medical images.

Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-04-20T13:30:09Z No. of bitstreams: 1 FelipeAlbuquerqueMarquesTese2015.pdf: 5827285 bytes, checksum: 3dfbabb913ebff137b6f41b2cd1ba5fc (MD5)<br>Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-04-20T13:30:28Z (GMT) No. of bitstreams: 1 FelipeAlbuquerqueMarquesTese2015.pdf: 5827285 bytes, checksum: 3dfbabb913ebff137b6f41b2cd1ba5fc (MD5)<br>Made available in DSpace on 2017-04-20T13:30:28Z (GMT). No. of bitstreams: 1 FelipeAlbuquerqueMarquesTese2015.pdf: 5827285 bytes, checksum: 3dfbabb913ebff137b6f41b2cd1ba5fc (MD5) Previous issue date: 2015-09-25<br>Genetic diseases of the skeleton affect the genesis of skeletal system. They are caused by mutations in genes which act on the cartilage and/or growth plate. The current classification of skeletal anomalies describes more than 456 distinct phenotypes organized into 40 groups. Of this total, 360 phenotypes are associated with defects in 336 genes (thus, 90 diseases remain to have their cause elucidated). The development of high resolution techniques for genomic analysis has enabled more genetic diseases, including skeletal phenotypes, to have molecular basis clarified. This research aimed to identify genes or regions in the human genome associated with genetic diseases of the skeleton. To this end, a pipeline was developed involving experiments and data analysis. Investigation of single nucletoide variation (SNV) was carried out using whole exome sequencing (WES) and submicroscopic structural variations were analyzed by Chromossomal Microarray Analysis (CMA). Moreover, Sanger sequencing, Fluorescence in situ Hybridization, in vitro functional tests (cell culture, qRT-PCR, Western Blot, Immunofluorescence and transcriptome), Immunohistochemistry and histochemistry were employed. 14 patients with rare diseases of skeleton (Craniosynostosis, s. FATCO sindrome, Catel-Manzke-like sindrome, Nager Syndrome and Rodriguez Syndrome) were selected. Craniosynostosis: of three patients, two had their molecular diagnoses elucidated, one with mutation in FGFR3 and the other with a translocation involving chromosomes 17q and 20q. s. FATCO: it wasn't possible to identify the causative mutation for this disease. Catel-Manzke-like Syndrome: initially this patient was diagnosed as Catel-Manzke Syndrome, and there was found a mutation in EXT2. Thus, this patient was reclassified as a new syndrome recently reported as seizures-scoliosis-macrocephaly. Nager and Rodriguez Syndrome: two of four have been diagnosed with mutation in SF3B4. For these patients, the results of qRT-PCR, Western Blot and Immunofluorescence together suggested that the phenotype is caused by SF3B4 haploinsuficiency. Immunohistochemistry and Histochemistry showed the expression of SF3B4 in cartilage tissue and the disorganization of hypertrophic cells in growth plate, respectively. The transcriptome result from cartilage tissue of one patient with SF3B4 mutation showed 12 underexpressed genes involved in skeletogeneses. The combination of techniques like classical cytogenetics and molecular cytogenetics as well as sequencing and in vitro assays were effective to achieve a diagnosis. Although there was an investigative core common to all diseases, investigations were customized to each case, seeking greater efficiency in the detection of the molecular basis and cost optimization of molecular research.<br>As doen??as gen??ticas do esqueleto s??o anomalias que envolvem a g??nese do sistema esquel??tico, causadas por altera????es em genes que atuam principalmente na cartilagem e/ou no n??cleo de crescimento. Na classifica????o atual h?? 456 doen??as do esqueleto categorizadas em 40 grupos. Destes, 360 patologias esquel??ticas est??o associadas a defeitos em 336 genes (portanto, existem 90 displasias esquel??ticas sem causa definida). O surgimento de t??cnicas de alta resolu????o de an??lise gen??mica tem permitido que cada vez mais doen??as gen??ticas, incluindo as doen??as gen??ticas do esqueleto, possam ter suas bases moleculares elucidadas. Esta pesquisa teve como objetivo a identifica????o e caracteriza????o de genes e regi??es do genoma humano associado a doen??as gen??ticas raras do esqueleto. Para isso, implantou-se um pipeline envolvendo experimentos e an??lise de dados. Esta tese investigou varia????es de nucleot??deo ??nico (SNVs) pelo uso Whole Exome Sequencing (WES) e varia????es estruturais submicrosc??picas, pelo uso Chromossomal Microarray Analysis (CMA). Al??m disso, fez-se uso de Sequenciamento de Sanger, Fluorescence in situ Hybridization, testes funcionais in vitro (cultura celular, qRT-PCR, Western Blot, Imunofluoresc??ncia e Transcritoma), Imunohistoqu??ca e histoqu??mica. Foram selecionados 14 pacientes com doen??as raras do esqueleto (Craniossinostose, s??ndrome FATCO, s??ndrome Catel-Manzke-like, s??ndrome de Nager e s??ndrome Rodriguez). Craniossinostose: dos quatro pacientes, dois foram diagnosticados com muta????o em FGFR3 e outro com uma transloca????o envolvendo os cromossomos 17q e 20q. S??ndrome de FATCO: n??o foi poss??vel identificar as bases moleculares da doen??a. ???Catel-Manzke-Like???: inicialmente diagnosticado com Sindrome de Catel-Manzke, o paciente teve uma muta????o detectada em EXT2, sendo reclassificado como uma nova s??ndrome (S??ndrome Convuls??o-Escoliose-Microcefalia). S??ndromes de Nager e Rodriguez: foram diagnosticado muta????es no gene SF3B4 em dois dos quatro pacientes. Nestes mesmos pacientes foram realizados qTR-PCR, Western Blot e Imunofluoresc??ncia que juntos sugeriram que estas patologias sejam causadas pela haploinsufici??ncia do SF3B4. A imunohistoqu??mica e histoqu??mica mostraram a express??o de SF3B4 na placa de crescimento e desorganiza????o de condr??citos hipertr??ficos, respectivamente. O transcritoma de um dos pacientes, com muta????o em SF3B4, evidenciou 12 genes ligados a esqueletog??nese com express??o diminu??da no tecido cartilaginoso. A combina????o de t??cnicas seja citogen??tica cl??ssica, citogen??tica molecular, sequenciamento, bem como an??lises in vitro se mostraram eficientes para se alcan??ar o diagn??stico. Embora houvesse um cerne investigativo em comum a todas as doen??as, as investiga????es foram personalizadas para cada caso, visando maior efici??ncia na detec????o das bases moleculares e otimiza????o dos custos da investiga????o molecular.

Francisella tularensis is a highly virulent zoonotic pathogen that is the causative agent of tularemia in humans. Two subspecies of F. tularensis are the most virulent in humans: tularensis (type A) and holarctica (type B), with less than 10 organisms via aerosol of a type A strain having the ability to cause fatal infection. Over the last decade much research has been done on the pathogenesis of this unique intracellular bacterium and many different virulence factors have been identified. The goal of this dissertation has been to identify and characterize the capsule-like complex (CLC) surface antigen of F. tularensis, and to determine its role in virulence and immunoprotection in a mouse model. In addition, I have investigated the role of CLC in biofilm formation. The CLC appears as a negatively staining material surrounding F. tularensis cells during transmission electron microscopy (TEM). I found that the CLC in the type B live vaccine strain (LVS) could be significantly diminished by deleting two glycosyl transferase genes (LVSΔ1423-22) in the putative polysaccharide locus, FTL_1432-FTL_1421. In addition, I determined that the CLC was not a typical polysaccharide capsule, but was in fact composed of over 50 proteins and glycoproteins including known virulence determinants, such as GroEL, DnaK, and ClpB. Upon further evaluation of the CLC, I determined that it was composed of an increase in production of outer membrane vesicles and tubules (OMV/T). These OMV/T appeared to be self-aggregating into what I visualized through TEM as the CLC. LVSΔ1423-22 was attenuated in the mouse model, and BALB/c mice immunized with CLC and adjuvant were protected against challenge with LVS. In addition to virulence, the CLC appears to play a role in biofilm formation and development. F. tularensis type B strains lacking the surface antigens CLC or CLC and O-antigen, develop a 2-7-fold more robust biofilm than the parent strains. The biofilm matrix contains a glucan-like EPS, proteins, and extracellular DNA, and further characterization may lead to determining if the biofilm acts as an environmental survival mechanism for F. tularensis. In summary, the CLC appears to be a novel surface antigen composed of upregulated OMV/T that is present in type A and B F. tularensis. Deficiency in CLC contributes to increased biofilm formation that could contribute to the survival of F. tularensis in a wide range of environmental niches. Furthermore, the CLC contributes to virulence of type B strains and elicits a protective immune response to type B challenge. A CLC-deficient type A strain could be a candidate for a new live vaccine strain, and therefore further investigation of such a mutant is warranted.<br>Ph. D.

PONTIFÍCIA UNIVERSIDADE CATÓLICA DO RIO DE JANEIRO<br>COORDENAÇÃO DE APERFEIÇOAMENTO DO PESSOAL DE ENSINO SUPERIOR<br>PROGRAMA DE SUPORTE À PÓS-GRADUAÇÃO DE INSTS. DE ENSINO<br>Esta pesquisa tem o objetivo de gerar entendimentos sobre duas escolas estaduais internas a duas instituições de medidas socioeducativas. Sob a ótica da metodologia participativa, o estudo se propõe a analisar os discursos produzidos por professores e alunos destas escolas. A investigação assume um caráter transdisciplinar ao se colocar no terreno da Sociolinguística Interacional e da Prática Exploratória. A partir dessa perspectiva, são analisadas transcrições de áudio das entrevistas realizadas com professores e alunos e as Atividades Pedagógicas com Potencial Exploratório, realizadas nas turmas do 3 , 5 e 9 anos da escola interna à unidade masculina. A análise é feita com o objetivo de: compreender como aconteceu a interação do pesquisador com os alunos; observar as estratégias de gerenciamento de face e estigma; investigar as crenças presentes nas narrativas dos professores sobre esse espaço de ensino; e, a partir dos puzzles gerados pelos alunos, entender os questionamentos dos alunos e criar inteligibilidade sobre a qualidade de vida dessa escola. A metodologia bricoleur de fazer pesquisa permite o intercâmbio de teorias que, ao olhar para o mesmo corpus de dados, possibilita reconhecer a complexidade e fluidez dos mesmos, valorizando o caráter multicultural e dinâmico da sociedade contemporânea. Essa investigação possibilitou compreendermos as crenças dos participantes da pesquisa sobre aquele espaço de ensino de modo coletivo, situado e transitório, destacando-se: o encontro misto entre estigmatizados e não estigmatizados; as estratégias utilizadas pelos participantes das interações, a fim de preservar a face e diminuir o estigma; o caráter instável do estigma; a multiplicidade de discursos sobre esse local presente na análise das narrativas; bem como as reflexões dos alunos sobre a estrutura e o funcionamento da escola, suas trajetórias de vida e seus sentimentos e anseios.<br>This dissertation aims to understand a state school attended by students in conflict with the law. From the perspective of a participative methodology, the study focuses on the discourses produced by the teachers and students of this school. The investigation assumes a transdisciplinary approach by bringing together the fields of Interactional Sociolinguistics and Exploratory Practice. Within this framework, the recorded interviews of teachers and students were transcribed and analyzed. Also, the Pedagogical Activities with Exploratory Potential, performed in the 3rd, 5th e 9th classes of this Brazilian school were analyzed. The analytical work aims to understand how the researcher s interaction with the students occurred; to observe the management strategies of face and stigma; to investigate the beliefs present in the teachers narratives about this educational context; and, based on the students puzzles about their school, understand their puzzlement and create intelligibility about this school s quality of life. The bricoleur research methodology adopted allows the use of a combination of theories to interpret the same data from various perspectives. Thus, enabling the recognition of data complexity and fluidity, as well as valuing the multicultural and dynamic character of contemporary society. This investigation made it possible to understand the beliefs of the research participants about their teaching context in a collective, situated and transitory way, highlighting: the mixed encounterbetween those stigmatized and non-stigmatized; the strategies used by the participants of the interactions in order to preserve each other s face and reduce stigma; the unstable character of stigma; the multiplicity of discourses about this place present in the narrative analysis; and the students reflections about the school structure and functioning, their life trajectories, feelings and aspirations.

Internucleosomal DNA cleavage is often regarded as the biochemical hallmark of apoptosis and can be reproduced in vitro in rat liver nuclei. This chromatin cleavage in rat liver nuclei was further characterised and studies described in this thesis showed that the DNA was initially cleaved into 700kbp, 200-250kbp and 30-50kbp fragments via a Mg2+ dependent process which was potentiated by Ca2+. Further investigations of the DNA cleavage processes were carried out in primary hepatocytes using TGF-1 to induce apoptosis. The studies showed for the first time that treatment of hepatocytes with TGF-1 resulted in multi-step DNA cleavage as seen in rat nuclei. Cycloheximide and an interleukin-1-converting enzyme (ICE)-like inhibitor, benzyloxycarbonyl-Val-Ala-Asp-(OMe) fluoromethyl ketone (Z-VAD.FMK) and Z-Asp-Glu-Val-Asp-(OMe) fluoromethyl ketone (Z-DEVD.FMK), a potent inhibitor of CPP32, blocked DNA cleavage and apoptosis. During apoptosis there was a time dependent increase in CPP32-like proteolytic activity in lysates isolated from TGF-1 treated hepatocytes, which was detected with the fluorogenic assay using Z-DEVD-amino-trifluoromethyl-coumarin (Z-DEVD.AFC). This activity was abolished when hepatocytes were pre-treated with either Z-VAD.FMK, Z-DEVD.FMK or cycloheximide. Unlike cycloheximide, Z-VAD.FMK and Z-DEVD.FMK were potent inhibitors of activated lysate from TGF-1 treated hepatocytes. Immunoblotting showed the processing of pro-CPP32 to its active form. In conclusion, this study shows for the first time that TGFF-1 mediated apoptosis involves the activation of ICE-like proteases and that cycloheximide inhibits apoptosis by blocking upstream of this ICE-like activity.

Elevated levels of the cysteine protease cathepsin S (cat S) are found in Cystic fibrosis (CF) lung secretions, however, the role of cat S in CF lung disease is unclear. Cat S is capable of maintaining its activity at a neutral pH allowing it to remain active outside of the cell. Consequently, cat S has the capacity to promote remodelling of the extracellular matrix via its potent elastolytic activity. In addition, cat S can cleave and inactivate key antimicrobials in the CF airways. On the basis of findings to date, we hypothesise that active cat S contributes to the pathogenesis of CF lung disease and represents a viable therapeutic target for the treatment of chronic lung disease. The βENaC transgenic mouse model recapitulates essential features of chronic CF lung disease such as airway mucus obstruction, inflammatory lung damage and increased levels of cat S activity in the lungs of the βENaC transgenic mouse compared to wild-type controls. Pharmacological knockdown of cat S activity was achieved in the βENaC mouse using the cat S inhibitor VBY-999. In this thesis we have demonstrated that inhibition of cat S reduces inflammatory cell infiltration into the lung as well as levels of pro-inflammatory cytokines in bronchoalveolar lavage fluid in both the early and late stage lung disease in the βENaC mouse model. Furthermore, concomitant reductions in mucus plugging were also observed. Late treatment with the cat S inhibitor had no effect on lung tissue damage, however, mucus plugging was reduced. We generated a βENaC mouse in which cat S was genetically deleted, the results from which supported our findings from the inhibitor study. These results support the hypothesis that active cat S plays a role in the pathogenesis of chronic lung disease and may be a viable and promising target in the treatment of chronic lung diseases such as CF and COPD.

El present treball es centra en la integració de processos innovadors per a la remoció biològica de Nitrogen i Sofre. La desnitrificació parcial ha sorgit com a procés innovador per actuar en sinergia amb el anammox, com una alternativa a la nitritació parcial. En el cas de la desnitrificació impulsada per sulfurs, la desnitrificació autotròfica parcial (PAD) és una altra alternativa valuosa. La integració de la PAD amb el procés anammox (PAD/A) és d'especial interès per als tractaments d'aigües residuals riques en compostos de nitrogen i sofre, com les derivades de refineries de petroli, curtiembres i processament de peix. En el PAD/A, la desnitrificació autotròfica s'ha d'operar de manera que converteixi el nitrat a nitrit, per proporcionar el nitrit requerit pel anammox; la dessulfuració de l'biogàs podria aconseguir en la mateixa unitat, com a alternativa sostenible als tractaments fisicoquímics convencionals. En aquest marc, la present tesi va tenir com a objectiu aprofundir en l'aplicació de l'innovador procés PAD/A, en aigües residuals d'alta càrrega en nitrogno i sofre. El anammox i la PAD s'han estudiat en línies de treball paral·leles, abordant diferents qüestions relacionades amb els dos tractaments innovadors. Les aigües residuals de curtiembres es van seleccionar com a target per al PAD / A i es va estudiar la inhibició de la biomassa anammox. Es va aconseguir una ràpida posada en marxa d'un reactor gas-lift anammox amb càrregues volumètriques fins 0,48 gN-NO2-/l/d. Es va observar la precipitació mineral en els grànuls després de 130 dies d'operació. Es va realitzar un estudi extens per caracteritzar la composició de l'precipitat, així com el seu impacte en el rendiment de l'reactor, l'activitat de la biomassa i la població microbiana. Els resultats ofereixen coneixements útils per a l'estabilitat a llarg termini de sistemes granulars anammox. El possible efecte inhibidor dels compostos biorrefractarios i la salinitat de les aigües residuals de curtiembres es va avaluar mitjançant experiments en batch. Els resultats revelen que no es va poder atribuir una inhibició evident als compostos biorefractarios, mentre que es va observar una influència moderada deguda a el contingut de salinitat, que es pot abordar mitjançant una adequada aclimatació de la biomassa. Es conclou que no hi ha cap limitació tècnica per a l'aplicació de l'procés anammox en les aigües residuals de curtiembres. El PAD es va estudiar en un reactor CSTR, alimentat amb nitrat i sulfur. El treball va avaluar l'efecte de les ràtios S/N de l'influent i les condicions de SRT sobre el rendiment de l'acumulació de nitrits. Es va observar una acumulació reeixida de nitrit després d'una setmana de condició estable de limitació de S i es van assolir alts nivells de nitrit durant tot el treball experimental. En totes les fases experimentals es va observar una eficiència d'acumulació de nitrit de l'70-100%. Es va observar un clar canvi de població microbiana: el gènere Sulphurimonas va mostrar una abundància relativa de l'90% en l'inòcul i va ser superat gairebé del tot pel gènere Thiobacillus, després de 80 dies d'operació. S'especula que l'eficiència bioenergètica microbiana és una possible raó subjacent de l'canvi de població. Es va realitzar un estudi de termodinàmica per estudiar el canvi catabòlic observat. Els resultats indiquen que tot i que la desnitritación és una reacció molt favorable, la desnitratación podria ser una resposta energètica valuosa en cas de limitació de l'donador d'electrons. La estequiometria catabòlica de el procés es presenta de manera agregada com a eina de suport per al disseny i l'optimització de l'complex procés de desnitrificació autrotrófica, ja que ofereix totes les reaccions intermèdies i les possibles combinacions d'e-donador i e-acceptor.<br>El presente trabajo se centra en la integración de procesos novedosos para la remoción biológica de Nitrógeno y Azufre, específicamente en el proceso de anammox y la desnitrificación autotrófica a base de sulfuros. La desnitrificación parcial ha surgido como proceso innovador para actuar en sinergia con el anammox, como una alternativa a la nitritación parcial. En el caso de la desnitrificación impulsada por sulfuros, la desnitrificación autotrófica parcial (PAD) es otra alternativa valiosa. La integración de la PAD con el proceso anammox (PAD/A) es de especial interés para los tratamientos de aguas residuales ricas en compuestos de nitrogeno y azufre, como las derivadas de refinerías de petróleo, curtiembres y procesamiento de pescado. En el proceso PAD/A, la desnitrificación autotrófica debe operarse de manera que convierta el nitrato en nitrito, para proporcionar el nitrito requerido por el anammox; la desulfuración del biogás podría lograrse en la misma unidad, como alternativa sostenible a los tratamientos físico-químicos convencionales. En este marco, la presente tesis tuvo como objetivo profundizar en la aplicación del novedoso proceso PAD/A, en aguas residuales de alta carga en nitrogno y azufre. El anammox y la PAD se han estudiado en líneas de trabajo paralelas, abordando diferentes cuestiones relacionadas con los dos tratamientos innovadores. Las aguas residuales de curtiembres se seleccionaron como target para el PAD/A y se estudió la inhibición de la biomasa anammox. Se logró una rápida puesta en marcha de un reactor gas-lift anammox con cargas volumétricas hasta 0,48 gN-NO2-/l/d. Se observó la precipitación mineral en los gránulos después de 130 días de operación. Se realizó un estudio extenso para caracterizar la composición del precipitado, así como su impacto en el rendimiento del reactor, la actividad de la biomasa y la población microbiana. Los resultados ofrecen conocimientos útiles para la estabilidad a largo plazo de sistemas granulares anammox. El posible efecto inhibidor de los compuestos biorrefractarios y la salinidad de las aguas residuales de curtiembres se evaluó mediante experimentos en batch. Los resultados revelaron que no se pudo atribuir una inhibición evidente a los compuestos biorefractarios, mientras que se observó una influencia moderada debida al contenido de salinidad, que se puede abordar mediante una adecuada aclimatación de la biomasa. Se concluye que no existe ninguna limitación técnica para la aplicación del proceso anammox en las aguas residuales de curtiembres. El PAD se estudió en un reactor CSTR, alimentado con nitrato y sulfuro. El trabajo evaluó el efecto de los ratios S/N del influente y las condiciones de SRT sobre el rendimiento de la acumulación de nitritos. Se observó una acumulación exitosa de nitrito después de una semana de condición estable de limitación de S y se alcanzaron altos niveles de nitrito durante todo el trabajo experimental. En todas las fases experimentales se observó una eficiencia de acumulación de nitrito del 70-100%. Se observó un claro cambio de población microbiana: el género Sulphurimonas mostró una abundancia relativa del 90% en el inóculo y fue superado casi por completo por el genero Thiobacillus, después de 80 días de operación. Se especula que la eficiencia bioenergética microbiana es una posible razón subyacente del cambio de población. Se realizó un estudio de termodinámica para estudiar el cambio catabólico observado. Los resultados indican que a pesar de que la desnitritación es una reacción muy favorable, la desnitratación podría ser una respuesta energética valiosa en caso de limitación del donador de electrones. La estequiometría catabólica del proceso se presenta de forma agregada como herramienta de apoyo para el diseño y la optimización del complejo proceso de desnitrificación autrotrófica, ya que ofrece todas las reacciones intermedias y las posibles combinaciones de e-donador y e-aceptor.<br>The present work focuses on the integration of novel processes for the biological removal of Nitrogen (N) and Sulphur (S), specifically the anammox process and the sulphide-based autotrophic denitrification. In the recent years, partial denitrification (PD), i.e. the reduction of nitrate to the stage of nitrite typically over the oxidation of an organic electron donor, has emerged as a possible innovative process to act in synergy with anammox, as an alternative to partial nitritation whose long-term stability is reported to be critical in many applications. In case of sulphide-driven denitrification, partial autotrophic denitrification (PAD) is another valuable alternative. The novel integration of PAD with the anammox process (PAD/A) is of particular interest for the treatments of wastewaters highly rich in nitrogen and sulphur compounds, such as those derived from petroleum refineries, tanneries and fish processing. In the perspective of PAD/A implementation, S-driven denitrification should be operated in such a way to convert nitrate to nitrite only, in order to provide the nitrite supply required by the anammox biomass. Besides, biogas desulphurization could be accomplished in the same unit, as a sustainable alternative to the typically adopted chemical-physical treatments. Within this framework, the present thesis aimed at gaining insight on the application of the novel PAD/A process, in high-strength wastewaters. The anammox and the PAD process have been studied in parallel lines of work, addressing different issues related to the two innovative treatments. Industrial wastewater originated from vegetable tannery industry was selected as target stream for PAD/A implementation and the potential inhibitory drawbacks on the anammox granular biomass were studied. A fast start-up of an anammox gas-lift reactor was achieved with volumetric loads up to 0,48 gN-NO2-/l/d. Mineral precipitation on the granules&#8217; surface was observed after 130 days of operation. An extensive study was conducted in order to characterise precipitate composition as well as its impact on reactor performance, biomass activity and microbial population. The possible inhibitory effect of bio-refractory compounds and high salinity of vegetable tannery wastewater was assessed through batch test experiments. The outcomes revealed that no evident inhibition could be ascribed to the organic tannins-related fraction of the industrial matrix, whereas a moderate disturbance was observed for the salinity content, that can be addressed through proper biomass acclimation. Thereby, it is concluded that there is no technical limitation for the application of anammox process on vegetable tannery wastewaters. The PAD process was studied in a CSTR reactor fed by nitrate and sulphide. The work aimed at evaluating the effect of influent S/N ratios and SRT conditions on nitrite accumulation. Successful nitrite accumulation was observed after one week of stable and strict S-limiting condition and maintained throughout the experimental work. Nitrite accumulation efficiency of 70-100% and nitrite conversion efficiency of 70% were observed at all the experimental phases, calculated over the converted and influent nitrate, respectively. A clear microbial population shift was observed: Sulfurimonas showing 90% relative abundance in the seeding sludge was almost completely out-competed by Thiobacillus, after 80 days of operation. It is speculated that microbial bioenergetics is a possible underlying reason of the clear population shift. A thermodynamic-based study was conducted in order to study the catabolic change observed in the experimental work, i.e. from full denitrification to partial denitrification (denitratation). Results indicate that even though denitritation is a highly favourable reaction, the sole denitratation could be a valuable energetic response in case of electron donor limitation. Aggregated information on process catabolic stoichiometry are presented as support tool for process design and optimization of the complex S-based denitrification.<br>Universitat Autònoma de Barcelona. Programa de Doctorat en Ciència i Tecnologia Ambientals

The multicatalytic proteinase (MCP) is a 700 kDa multisubunit complex which is found in eukaryotic cells. MCP is involved in ubiquitin-dependent non-lysosomal protein degradation and has been implicated in the processing of antigens by the major histocompatability complex class 1 pathway. The MCP purified from rat liver is composed of at least 16 distinct subunits of molecular masses between 22 and 34 kDa. MCP is known to possess at least three distinct peptidase activities, described as the 'trypsin-like', 'chymotrypsin-like' and 'peptidylglutamylpeptide hydrolase' activities. However, it is not known which subunits are responsible for the different peptidase activities. The main aims of my study were (1) to establish which subunit(s) possesses the active site responsible for 'trypsin-like' activity, (2) to identify the catalytic amino acid residues responsible for this activity and (3) to determine whether all MCP particles possess these components. 'Protection' experiments with the peptide aldehyde leupeptin, which is a specific reversible inhibitor of the 'trypsin-like' activity, have identified one or two subunits which possess thiol groups essential for 'trypsin-like' activity. These thiol groups are probably not involved in catalysis but may be located very close to the active site responsible for 'trypsin-like' activity. An active site specific peptidyl chloromethane which specifically inactivates the 'trypsinlike' activity, labels two distinct subunits of 23-24 kDa. One of these polypeptides is also labelled by an active site specific peptidyl diazomethane for the 'chymotrypsin-like' activity, in addition to three other distinct polypeptides of 28-23 kDa. Together these results suggest that one active site is responsible for 'trypsin-like' activity while four distinct active sites are responsible for 'chymotrypsin-like' activity. Affinity chromotography was carried out using an immobilised specific inhibitor of the 'trypsin-like' activity. The results of these studies suggest that all MCP particles purified from rat liver possess the catalytic component responsible for the 'trypsin-like' activity.

Thesis (M.A.)--University of Kentucky, 2002.<br>Title from document title page. Document formatted into pages; contains vii, 63 p. : ill. Includes film clips utilizing MPG files. Includes abstract. Includes bibliographical references (p. 60-62).

Plk4 has been termed a `suicide kinase' because it promotes its own destruction to regulate protein levels. We identified numerous autophosphorylated residues within a region of Plk4 called the Downstream regulatory element (DRE). We find that phosphorylation of a single residue is sufficient for Slimb recruitment and phosphorylation of the surrounding residues builds a high affinity Slimb-binding site. These autophosphorylation events are dependent on Plk4 homodimerization, although the domains that mediate this dimerization are unknown. We show that Plk4 homodimerization is mediated by interactions between the PB1-PB2 cassette. We find that like all Polo kinases, Plk4 encodes a mechanism of kinase autoinhibition. Unlike other Polo kinases, which rely on external inputs for relief of inhibition, Plk4 is self-sufficient in relieving kinase inhibition. This relief of autoinhibition is regulated by PB3 of Plk4 and is dependent on homodimerization, thereby making homodimerization a necessary step in formation of the Slimb phosphodegron on Plk4. Polo Boxes are known as multifunctional domains, and the Polo Boxes of Plk4 are no different. We identified numerous Slimb-mediated ubiquitination sites on PB1 as well as PB2. Furthermore, the PB1-PB2 cassette mediates the interaction between Plk4 and the N-terminus of Asterless. In Drosophila cells, Plk4 requires Asterless for centriolar localization and Asterless overexpression drives centriole amplification in a Plk4 dependent manner. This is a fascinating result as endogenous Plk4 protein levels are undetectable in S2 cells, making it hard to envision a scenario where overexpression of Asterless could shuttle a non-existent Plk4 population to the centriole to initiate duplication. We found that in addition to shuttling Plk4 to the centriole, Asterless stabilizes Plk4, likely protecting Plk4 at the centriole to allow it to `license' the centriole for duplication. Moreover, we show that Asterless encodes two distinct Plk4 binding sites: the previously described N-terminal binding site as well as a novel C-terminal binding site. We found that the interaction between the C-terminal of Asterless and Plk4 is necessary for centriole duplication while the interaction between the N-terminal of Asterless and Plk4 is expendable. Together these findings provide significant insight into Plk4 biology and the mechanisms which limit its activity.

Error Propagation Analysis (EPA) is a technique for understanding how errors affect a program’s execution and result in program failures. For this purpose, EPA usually compares the traces of a fault-free (golden) run with those from a faulty run of the program. This makes existing EPA approaches brittle for multithreaded programs, which do not typically have a deterministic golden run. In this thesis, we study the use of likely invariants generated by automated approaches as alternatives for golden run based EPA in multithreaded programs. We present Invariant Propagation Analysis (IPA), an approach and a framework for automatically deriving invariants for multithreaded programs, and using the invariants for EPA.We evaluate the invariants derived by IPA in terms of their coverage for different fault types across six representative programs through fault injection experiments. We find that stable invariants can be inferred in all six programs, although their coverage of faults depends on the application and the fault type.<br>Applied Science, Faculty of<br>Electrical and Computer Engineering, Department of<br>Graduate

Since their discovery, subglacial lakes beneath the Antarctic ice sheet have been a focus of scientific curiosity and study. It has been postulated that subglacial lakes present a viable habitat for microbial life and are underlain by sediments containing a record of changes in the ice sheet above. Subglacial Lake Ellsworth, West Antarctica, is the subject of a continued project aiming to achieve clean access in order to explore these hypotheses. One of the key outcomes of lake sampling is to establish the hydrochemistry, from which deductions about the limnology of the system can be made. A new theoretical insight into the hydrochemistry of subglacial lakes is achieved through reanalysis of the geochemical dataset published from meteoric and accretion ice from above Subglacial Lake Vostok, East Antarctica. It is shown that Subglacial Lake Vostok is a well-mixed system at steady-state. Of the flux of water to the lake, ~60% drains away from the lake and ~40% freezes at the ice-water interface. A downstream subglacial hydrological catchment is strongly indicated. An average cationic denudation rate of ~9meq∑+·m-2·yr-1 is inferred at the lake-sediment interface. The methods and outcomes of the Subglacial Lake Vostok model form the theoretical basis to scope the controls on the hydrochemistry of Subglacial Lake Ellsworth. A hydrologically open lake system leads to dilute, isotopically heavier water. A hydrologically more isolated lake system results in more concentrated waters with a stable water isotope composition that is removed from the local meteoric water line. Geochemical weathering reactions are expected to contribute the majority of dissolved ionic species and meteoric ice is shown to deliver oxygen and nitrogen to the lake. The model results provide a theoretical framework for the interpretation of analyses from lake water samples. The model results are also used to guide a sampling and analysis strategy which would optimise the scientific gains of subglacial lake sampling. The precise nature of the processes within Subglacial Lakes Ellsworth and Vostok remain untested, but this thesis provides a robust basis for interpreting the hydrochemistry of both lakes.

The spermatozoon is a highly complex cell which was thought to be transcriptionally inactive and solely delivers the paternal genome, centrioles and proteins to the oocyte. During the past few years, a large number of different sperm RNA species have been discovered, however, the functionality and the gene expression networks they may be involved remain unclear to date. Different studies showed that sperm RNAs provide a historical record of gene activity during spermatogenesis and that they are delivered to the oocyte. The studies reported herein had the following foci: Firstly, the spermatozoal transcriptomes of bovine, ovine, porcine and human were compared to their respective testis transcriptomes. Secondly, the spermatozoal transcriptome of each species was analysed and compared to identify shared gene expression networks that may be indicative of common functionality in the processes of spermatogenesis, fertilisation and reproduction. The results revealed common functional pathways, indicating a possible post-fertilisation role in the developing embryo. In addition, bioinformatic analysis revealed 23 mutually shared transcripts. To further characterise the potential transfer to the oocyte and to investigate the potential function and stability of these paternal transcripts, 16 transcripts were selected and followed in the developing bovine embryo. No clear potential functions for spermatozoal RNAs post-fertilisation and during embryo development could be derived from these experimental data. Assisted reproductive processes and the breeding industry rely on freezing spermatozoa and choosing the best candidate spermatozoon. Therefore as an additional focus, RNA profiles of frozen and fresh spermatozoa from the same donor were analysed and did not reveal major changes in the spermatozoal transcriptome caused by cryopreservation processes. Additionally as a last focus, the transcriptome of hyaluronic acid (HA) selected spermatozoa was compared to unselected spermatozoa and revealed no significant differences in RNA expression. However, a trend towards an increased expression of MOSPD3 was observed and investigated as a potential fertility marker. Expression of MOSPD3 protein was seen to be upregulated in motile spermatozoa compared to less motile by both Western Blot analysis and immunocytochemistry. The present data supports the suggestion that sperm binding to HA may represent more viable populations and that MOSPD3 is a potential marker of spermatozoa viability that could be developed into a diagnostic tool.

Adult hemoglobin (HbA) is made up of two pairs of globin chains. Some rare mutations of the globin chains can result in high affinity towards hemoglobin molecule thus changing the equilibrium of normal oxygen loading in lungs and the delivery of same to the tissues. Because of change in the affinity to the oxygen these mutations can result in erythrocytosis (polycythemia). Here we discuss a case of Familial Polycythemia likely due to novel hemoglobin variant. 42-year Caucasian male presents to the clinic with high hemoglobin for several years but otherwise denies any symptoms of headache, vision changes, chest pain. He had history of multiple phlebotomies. His past medical history is significant for Polycythemia, PICC line associated clot,Type2 Diabetes, Hypertension, Epidural abscess. Social history is significant for smokeless tobacco but otherwise non-smoker, non-alcoholic, not an IVDA and doesn’t use testosterone. Family history is significant for his sister, her two 14year old daughters and multiple other family members with elevated hemoglobin and undergo phlebotomies, maternal grandfather died of cancer in his 30s. Physical examination is only significant for BMI of 32.46 otherwise no skin discoloration or cyanosis. Laboratory data WBC 9.4 with normal differential, hemoglobin 18.2, hematocrit 55.4, platelet count 180,000, peripheral blood smear was within normal limits, negative for JAK-2, normal erythropoietin, negative for hemochromatosis, Oxygen dissociation p50 of 19 which is low indicating left shifted dissociation curve, hemoglobin electrophoresis HbA: 61.2%, HbA2: 3%, HbF: 0%, Beta variant: 35.8%. In order to further characterize beta variant Bi-directional sequence analysis for Molecular alterations was performed and the following alterations were detected Gene: HBB, DNA change: Codon 39, heterozygous CAG>CCG Protein change: P.G1n39Pro. [glutamine (Q) to proline (P)]. HGVS: c.119A>C, p.Q40P, Classification: Likely deleterious variant. (GenBank accession number NM_000518.4). This is a previously unreported beta chain hemoglobin variant present. This hemoglobin variant is named as Hemoglobin Hyden based on the place where this is found in Hyden, Kentucky. There have been four variants reported at codon 40 of the beta globin gene, which is an external contact site between beta globin and alpha-2 globin. One variant, Hb vassa, is associated with mild hemolytic anemia. The three other variants (Hb Alabama, Hb Tianshui and Hb San Bruno) are not associated with clinical or hematological abnormalities. In our opinion p.Q40P is likely a cause of erythrocytosis. In order to further establish the causality it may be beneficial to test first degree relative to in this family in order to determine whether the p.Q40P alteration tracks with disease and is not present in unaffected individuals. Hemoglobin threshold for phelebotomy to lower the risk of thrombosis and cardiovascular events is yet to be defined.

In questa tesi vengono mostrati i risultati delle prove di compressione, trazione e flessione effettuate su culmi di bambù di provenienza italiana. Le specie analizzate sono due e sono la Phyllostachys Edulis (MOSO) e la Phyllostachys Viridiglaucescens. Le prove sono state effettuate seguendo le norme ISO 22156 e ISO 22157, le uniche norme internazionali relative al bambù strutturale. Dai risultati ottenuti, sono stati successivamente definiti i valori caratteristici di resistenza a compressione e flessione dei culmi delle due specie italiane analizzate. Inoltre, è stato ideato e testato in laboratorio un nuovo e versatile collegamento per le strutture in bambù. Il collegamento, realizzato mediante un cilindro in legno solidarizzato con viti autofilettanti, è stato progettato in modo tale da essere prontamente impiegabile sia per la realizzazione di capriate sia per le strutture reticolari spaziali. La scelta di questa tipologia di collegamento è stata motivata da due aspetti principali. Il primo aspetto è che l’utilizzo di viti autofilettanti non richiede particolari capacità da parte del costruttore. Il secondo aspetto risiede nel fatto che per il fissaggio delle viti è richiesto un preliminare foro accomodante, il quale garantisce l’integrità del culmo durante l’avvitamento, evitando così spaccature longitudinali della canna di bambù. Nella seconda parte della tesi viene trattato il problema dell’instabilità lineare (linear buckling) per le aste sottoposte a sforzi di compressione, con la particolarità che la rigidezza flessionale sia variabile lungo l’asse longitudinale, caratteristica peculiare dei culmi di bambù. Primariamente viene esposta una breve introduzione storica del problema e, successivamente, viene proposto un metodo per la determinazione della soluzione approssimata in forma chiusa dell’equazione differenziale che regola il problema. Il metodo consiste nel rappresentare l’effettiva rigidezza flessionale, descritta da un polinomio di quinto grado, con una funzione lineare. Questo metodo permette altresì di valutare il carico critico euleriano attraverso una semplice formula, risultando così agevole per scopi professionali. Il metodo proposto, inoltre, permette di valutare approssimativamente in forma chiusa la deformata trasversale delle aste di bambù sottoposte a carico assiale eccentrico e in presenza di imperfezioni.<br>In this thesis the results of compression, tensile and bending tests on Italian bamboo culms are discussed. The analyzed bamboo are of two species, namely Phyllostachys Edulis (MOSO) and Phyllostachys Viridiglaucescens. These tests are carried out following the rules of ISO 22156 and ISO 22157. The characteristics values of the compressive and flexural strength of Italian bamboo culms are determined. A new and versatile connector for bamboo structures is developed and tested to evaluate its effectiveness. This connector is realized by wood parts and screws and it is appropriate for constructing trusses and space frames. The use of the screws to connect wood and bamboo is due to two main aspects. First, the screws are easy to use and no particular skill is needed. Second, no splitting arises along the culm since a preliminary drilling is needed to introduce the screw. In the second part of the thesis, the linear buckling problem of non-uniform columns is treated. Firstly, a brief overview of the history of the linear buckling is described. Since bamboo culms present non-uniform flexural stiffness along the length, an approach to evaluate the approximate analytical solution of the governing equation is presented. This method can calculate the critical load in a simple way, directly accessible to the designers. The proposed method gives a closed-form estimate of the bamboo column deflection, subjected to an eccentric axial load and initial curvature (crookedness).

Patients with inflammatory or infectious conditions such as periodontitis, peri-implantitis, osteomyelitis, rheumatoid arthritis, septic arthritis and loosened joint prosthesis display varying severity of destruction in the adjacent bone tissue. Bone loss in inflammatory diseases is considered a consequence of cytokine induced RANKL and subsequent enhanced osteoclast formation. Hence, osteotropic cytokines and their receptors have been suggested to be important for the pathogenesis of inflammation-induced osteolysis. It is, here, suggested that bacterial components, so called “pathogen associated molecular patterns=PAMPs”, may also be involved. Varieties of cells express receptors for PAMPs, including Toll-like receptors (TLRs) which are the first line of defence in the innate immune system. LPS (lipopolysaccharide), fimbria and lipoproteins from pathogenic bacteria such as P. gingivalis, S. aureus are ligands for TLR2 and flagellin from pathogenic flagellated bacteria like S. typhimurium is a ligand for TLR5.   Since the susceptibility to, or the severity of inflammation-associated bone diseases are likely related to differences in the tissue response, and the mechanisms by which PAMPs interact with bone cells are not fully understood, we aimed to elucidate the importance of different TLRs for inflammation induced bone loss by conducting in vitro and in vivo investigations. Activation of TLR2 and TLR5 in organ cultured mouse parietal bones increased bone resorption in a time- and concentration-dependent manner by a process inhibited by OPG and bisphosphonate, showing the crucial role of RANKL-induced osteoclast formation. In addition, the number of osteoclasts, expression of osteoclastic genes and osteoclastogenic transcription factors were increased. In the bones and in osteoblasts isolated from the bones, TLR2 agonists increased the expression of RANKL without affecting OPG, while TLR5 activation resulted in enhanced RANKL and decreased OPG. Activation of both TLR2 and TLR5 stimulated the expression in both bones and osteoblasts of prostaglandins and pro-inflammatory cytokines, known to stimulate RANKL. By blocking the cytokines and prostaglandin, we showed that TLR2 and TLR5 induced bone resorption and RANKL expression are independent of these molecules. Activation of TLR2, but not TLR5, in mouse bone marrow macrophage cultures inhibited RANKL-induced osteoclast formation, an effect not observed in committed pre-osteoclasts. Local administration in vivo of TLR2 and TLR5 agonists on the top of mouse skull bones enhanced local and systemic osteoclast formation and bone resorption. Using knockout mice, we showed that the effects by LPS from P. gingivalis (used as TLR2 agonist) and flagellins (used as TLR5 agonists) are explicit for TLR2 and TLR5 ex vivo and in vivo, respectively. These data show that stimulation of TLR2 and TLR5 results in bone resorption in vitro and in vivo mediated by increased RANKL in osteoblasts and thus may be one mechanism for developing inflammatory bone loss. Interestingly, histological analyses of skull bones of mice treated locally with TLR2 and TLR5 agonists revealed that the bones not only reacted with locally increased osteoclastogenesis (osteoclast formation), but also with locally increased new bone formation. This was observed on both periosteal and endosteal sides of the bones, as well as in the bone marrow compartment. The formation of new bone was seen close to osteoclasts in some parts, but also in other areas, distant from these cells. The response was associated with active, cuboidal osteoblasts, extensive cell proliferation and increased expression of genes coding for bone matrix proteins and osteoblastic transcription factors. In conclusion, activation of TLR2 and TLR5 in osteoblasts results in bone loss associated with enhanced osteoclast formation and bone resorption, as well as with increased osteoblast differentiation and new bone formation, indicating that inflammation causes bone modeling. The data provide an explanation why LPS from P. gingivalis and flagellin from flagella-expressing bacteria can stimulate bone loss. Since TLR2 and TLR5 can be activated not only by bacterial components, but also by endogenous ligands produced in inflammatory processes, the data also contribute to the understanding of inflammation induced bone loss in autoimmune diseases. Hopefully, these findings will contribute to the development of treatment strategies for inflammation induced bone loss.

Eisenmangel ist ein weltweites Ernährungsproblem für Pflanzen und allen von Ihnen abhängigen Sekundärkonsumenten. Er reduziert den Ertrag, die Qualität und Produktivität von z.B. Kulturpflanzen, was wiederum zu Mangelerscheinungen beim Menschen führen kann. Die Nahrungsmittelforschung hat ein großes ökonomisches und wirtschaftliches Interesse daran, die Eisenverfügbarkeit und -Aufnahme der Pflanzen zu erhöhen. In der vorliegenden Arbeit wurde eine kleine Familie von fünf (VTL1-5) neuen potentiellen Eisentransportern in Arabidopsis thaliana identifiziert und charakterisiert und somit ein weiterer Baustein zu der Aufklärung der Eisenhomöostase hinzugefügt. Bei den fünf Transportern handelt es sich um CCC1-like Proteine, von denen vier (VTL1-3 und 5) eine eisenabhängig regulierte Expression zeigen. Für VTL1 kann mittels Protein-tag Markierung sehr überzeugend eine Lokalisation in der Vakuolenmembran und ein damit verbundener Eisentransport in die Vakuole, analog zur Funktion von VIT1, gezeigt werden. Da vermutlich ein knockout von VTL1 zur Embryo Lethalität führt und auch die vit1-1 Mutante einen sehr verkümmerten Phänotyp unter Eisenmangel zeigt, scheinen beide Proteine getrennt voneinander an verschiedenen Schlüsselpositionen im Eisenhaushalt zu wirken. Auch für VTL2 und VTL5 kann eine Lokalisation an der Vakuolenmembran und der damit verbundene Eisenimport postuliert werden. Die heterologe Expression aller drei Gene in ∆ccc1 Zellen führt zur Erhöhung der vakuolären Eisenkonzentration. Für VTL4 wird mittels Protein-tag Markierung eine Lokalisation an der Plasmamembran mit einer Exportfunktion vorgeschlagen. VTL3 und VTL4 heterolog exprimierende ∆ccc1 Hefe-Zellen weisen einen geringeren Eisengehalt als Kontrollzellen auf. Alle fünf Proteine sind in der Lage sowohl den Mutanten-Phänotyp der ∆ccc1 Hefe-Mutante, wie auch der Arabidopsis vit1-1 und nramp3/nramp4 Doppelmutante zu komplementieren. Anhand dieser Tatsachen konnte die Eisentransportfähigkeit bewiesen werden.<br>Iron deficiency is a worldwide nutritional problem for plants and in general all heterotrophic organisms. Iron deficiency reduces crop productivity, quality and yield, which in consequence lead to iron deficiency symptoms in humans. Because approximately 50 % of the global human caloric intake is derived from a cereal grain diet, it is not surprising that the emphasis of nutrition research is on uptake, transport and storage of iron in plants, specifically in seeds. In this doctoral thesis a small family of five potential iron transporters (VTL1-5) in Arabidopsis thaliana has been identified and characterized. These five transporters are CCC1-like proteins, four of which (VTL1-3, 5) show a pattern of iron-dependent expression. Through the use of a protein tag, VTL1 is shown to be localized on the vacuolar membrane and associated with import of iron into the vacuole. In this respect VTL1 displays an analogous function to VIT1. Since the knockout of VTL1 likely leads to embryo lethality and seedlings of the vit1-1 mutant display an ephemeral phenotype caused by iron deficiency, both proteins seem to uniquely influence the iron homeostasis. As for VTL1 both VTL2 and VTL5 are localized on the vacuolar membrane and catalyze iron import. The heterologous expression of all three genes in ∆ccc1 cells leads to an increased vacuolar iron concentration. In contrast, VTL3 and VTL4 may function as iron exporters and play possibly roles in xylem loading. This conclusion is supported by the facts that a mCherry-VTL4 signal is localized to the plasma membrane of tobacco leaf cells and that ∆ccc1 cells in which VTL3 and VTL4 are heterologous expressed, show a decreased iron content compared to control cells. All five proteins are able to complement the ∆ccc1 yeast mutant and the two Arabidopsis vit1-1 and nramp3/nramp4 mutant phenotypes, thereby demonstrating a function in iron transport.

This paper suggests that a critical mass of elements is forming in China which, if not better controlled, will lead to some form of political regime change. The paper will (a) elaborate on China's major problems, grouped into "backbone changes" and "catalysts," (b) provide a balance sheet of remedies the government has attempted or proposed to date, and (c) examine the remedies' relative success or failure. Among the "backbone changes": decentralization, corruption, the emergence of interest groups, the government's possible loss of legitimacy, people's increased exposure to procedural democracy, the increase in the number of students receiving a foreign education, the privatization of education and divisions within the Chinese Communist Party. The "catalysts" include: massive urban and rural unemployment, corruption, forced displacement and the gentrification of China's cities.

Approved for public release; distribution is unlimited<br>This thesis investigates and explores the factors and conditions that have led Iran to pursue its aggressive ambitions. It begins with an overview of Iran's history prior to the revolution, specifically during the Shah's reign, which became the impetus for an Islamic Revolution. It moves on to describe Iran's history its culture, its geographical location, political structure and economy to provide some insights about how these factors have helped shape its ambitions. It continues by investigating Iran's ambitions through three lenses-Defensive, Aggressive, and Unstable Iran. The consequences and the implications of Iran's wider ambitions on a regional and global scale are further explored, as well as the reactions and actions taken by the international community to contain Iran's ambitions. Lastly, the thesis talks about courses of actions that might convince Iran to back down from its aggressive ambitions and transform itself into a peaceful country that will contribute stability to the region as well as resume friendly relations with the international community.

Individuals sensitive to domestic cat allergen Fel d 1 experience a variety of symptoms including eye irritation, respiratory irritation, asthma, and severe respiratory distress. Fel d 1 is a protein produced in the saliva and on the skin of domestic cats. Previous studies have demonstrated that Fel d 1 adheres to clothing, upholstery, and human hair and has been found in non-cat environments in levels high enough to cause allergic reactions in sensitive individuals. In a general sense, two very different approaches have been adopted to study Fel d 1. One area of the literature focuses on the molecular biology of Fel d 1 and its functions at the cellular level. These studies hold long-term promise for an effective clinical response to this persistent allergen. An entirely separate literature focuses on immediate practical solutions that remove Fel d 1 from the domestic environment. Within this literature there has been minimal emphasis on the possibility that different fabrics may have different affinities for Fel d 1. Therefore, the affinity of Fel d 1 for different fabrics is the focus of this study. The findings from this study will be of use in reducing allergic reactions in sensitive individuals through the choice of appropriate fabrics in clothing and upholstery. Forty domestic household cats were chosen for this study. Each cat was rubbed, in a manner similar to petting, with an assembled fabric square based on a Latin-square design. Each Latin-square design consisted of a 6x6 fabric grid and included the fabrics silk dupioni, wool suiting, cotton denim, cotton damask, polyester suede and polyester knit. The random organization of the fabrics into the grid removed bias for the location of fabrics within the square during Fel d 1 collection. After rubbing, the Latin-square fabric block was disassembled and Fel d 1 was extracted from each fabric type and analyzed via quantitative ELISA.; The results were statistically analyzed with a univariate ANOVA. Fabrics significantly differ (pless than]0.001) in Fel d 1 retention and fall into three groups. Silk dupioni collected the least amount of Fel d 1. Wool suiting, cotton denim and cotton damask were intermediate in Fel d 1 collection, while polyester suede and polyester knit collected the highest amounts of Fel d 1. Samples were also collected for a time study to determine if Fel d 1 bound on fabric degrades, or otherwise diminishes, over time. 14 weeks (approximately 3 months) after collection, Fel d 1 was extracted from fabrics and quantified by ELISA. A paired T-test was used to evaluate changes in Fel d 1 levels on specific fabrics over the 14 week period. When compared to extractions performed immediately after exposure, the amount of Fel d 1 released from specific fabrics after 14 weeks was significantly reduced. From these studies I conclude that an individual allergic to Fel d 1 may be able to limit their allergen exposure by selecting fabrics less likely to collect the allergen for their environment. Natural fibers (silk, wool, and cotton) collected less Fel d 1 than polyester fabrics, suggesting that natural fibers are recommended over fabrics containing polyester for persons allergic to cats.<br>ID: 030423055; System requirements: World Wide Web browser and PDF reader.; Mode of access: World Wide Web.; Thesis (M.S.)--University of Central Florida, 2011.; Includes bibliographical references (p. 35-38).<br>M.S.<br>Masters<br>Biology<br>Sciences

This thesis describes the development of a rapid antistaphylococcal bioassay and chemical investigation of antistaphylococcal extracts derived from marine invertebrates collected from northern New South Wales. This thesis is divided into three sections. Chapter 2 describes the development of a novel HTS antistaphylococcal bioassay and the subsequent screening of a marine invertebrate extract collection using the developed assay. Chapters 3-7 report on the isolation and structure elucidation of natural products from antistaphylococcal extracts derived from three sponges and two bryozoans. Finally, in chapter 8, the isolated natural products were investigated to determine if they were active against Staphylococcus aureus. The MIC80 of active compounds were reported as was the bacteriostatic or bactericidal mode of action of these bioactive compounds.<br>Thesis (PhD Doctorate)<br>Doctor of Philosophy (PhD)<br>Griffith School of Environment<br>Science, Environment, Engineering and Technology<br>Full Text

The purpose of this study is to contribute to previous research on the subject of language transition in Tanzania. The aim is also that the information gained in this study can be used to improve students' prospects when they undergo the linguistic transition from primary school to secondary school. This is a qualitative study with quantitative elements, using method triangulation, which examines how students experience the linguistic transition from Kiswahili to English and how it affects them. The study also highlights the students' own opinions about which language they would prefer as the language of instruction in secondary school. Relevant information about the history of Tanzania and the linguistic situation in the country are outlined. Previous research on the subject is also presented. The two theories that are used are Bourdieu’s social theory and Said's theory of Orientalism. The study uses focus groups and surveys in oder to answer the questions at hand. The research itself is conducted in two government schools in Dar es Salaam in Tanzania. The three questions at issue are as follows: Do the students feel that they have sufficient knowledge of English in order to be ready for the transition of the language of instruction from Kiswahili to English in secondary school? How do the students perceive that the transition of languages affects their performance in school and in their future? If the students had the opportunity to choose, which language would they prefer as the language of instruction in Secondary school in Tanzania? The conclusions are that the students feel that the linguistic transition is difficult and that they lack sufficient knowledge of English. Although several of the students struggle with the vocabulary and the pronunciation, they feel that English is the path to higher education and a good job. These are contributing factors to the fact that the majority of the students prefer English as their language of instruction in secondary school, to their own national language, Kiswahili.<br><p>MFS-uppsats</p>

Upon its discovery in 2006, the young L7.5 companion to the solar analog HD 203030 was found to be approximate to 200 K cooler than older late-L dwarfs, which is quite unusual. HD. 203030B offered the first clear indication that the effective temperature at the L-to-T spectral type transition depends on surface gravity: now a well-known characteristic of low-gravity ultra-cool dwarfs. An initial age analysis of the G8V primary star indicated that the system was 130-400 Myr old, and so the companion would be between 12 and 31 M-Jup. Using moderate-resolution near-infrared spectra of HD. 203030B, we now find features of very low gravity comparable to those of 10-150 Myr old L7-L8 dwarfs. We also obtained more accurate near-infrared and Spitzer/IRAC photometry, and we find a (J - K) MKO color of 2.56 +/- 0.13 mag-comparable to those observed in other young planetary-mass objects-and a luminosity of log (L-bol/L-circle dot) = -4.75 +/- 0.04 dex. We further re-assess the evidence for the young age of the host star, HD 203030, with a more comprehensive analysis of the photometry and updated stellar activity measurements and age calibrations. Summarizing the age diagnostics for both components of the binary, we adopt an age of 100 Myr for HD 203030B and an age range of 30-150 Myr. Using cloudy evolutionary models, the new companion age range and luminosity result in a mass of 11 M-Jup with a range of 8-15 M-Jup, and an effective temperature of 1040 +/- 50 K.

The backprojection-then-filtering algorithm was applied to proton CT data to reconstruct a map of proton stopping power relative to water (RSP) in air, water and bone. Backprojections were performed along three commonly used path estimates for the proton: straight line path, cubic spline path, and most likely path. The proton CT data was obtained through simulations using the GEANT4 simulation toolkit. Two elliptical phantoms were inspected, and an accuracy of 0.2% and 0.8% was obtained for the RSP in water and bone respectively in the region of interest, while the RSP of air was significantly underestimated.

Thesis (MTech (Built Environment))--Peninsula Technikon, 2001<br>The construction industry globally, contributes between 18% and 24% of the GDP, and because of its labour intensive characteristics, contributes handsomely to total employment, forming important backward and forward linkages with the rest of the economy. Nevertheless, the extent and sophistication of these linkages crucially depend on the relative development of the construction industry relative to the overall economy. In the developing countries, these linkages are not very strong because of the use of informal materials, which is not commercialised and whose opportunity costs are often zero, and the huge imports of construction materials used in the modem sector of the economy. However, whether in the developed or developing economies, the construction industry is a major contributor to economic growth and development by providing the necessary infrastructure that facilitates production, consumption and recreational activities. In fulfilling these activities, the construction industry generates huge wastes of which only a tiny proportion are recycled and reused. However, in economies and countries where adequate and functional housing is a problem mainly due to lack of affordability, recycling and reuse of construction waste is a necessary prerequisite to enhancing housing affordability in these countries. This is the current situation that South Africa finds itself "''here because of its past history of 'apartheid', economic opportunities and amenities were denied to the blacks. There is nowhere that this deprivation is more pronounced than in the built environment sector where housing shortages and general disamenities prevail. High levels of unemployment further exacerbate the situation, - - which is a consequence of low skills and high illiteracy-rates. Thus, housing demand and supply by this group of the population are most likely, on the evidence available, to fall predominantly within the low-income housing category. Presently, all households falling into this category rely on financial assistance from the government to facilitate low-cost housing consumption because of pervasive poverty, which itself is due to the very high unemployment rate, illiteracy, lack of skills and general deprivation: a legacy of 'apartheid' policies enforced by previous government. The dilemma however is how to meet the huge housing demand within the limited resources available to the government on the one hand, and on the other, to satisfy such demand without compromising the environmental sustainability of the physical environment. Thus, the thesis aims to determine ways in which the construction industry could contribute to the sustainability of the carrying capacity of the biophysical environment and enhance social sustainability by facilitating affordability through the possible reductions to construction costs through recycling and reuse. By means of questionnaires and detailed interviews, underscored by a qualitative research approach, the potential of construction recycling and the possible contributions to environmental sustainability and housing affordability are determined. At completion, it is expected that this work will not only contribute to existing knowledge but would be of significance in terms of policy formulation to construction industry practitioners, central and local government policy makers, and other governmental and non-governmental organisations operating in the area of housing.

The case study research sought to investigate and establish the attitude of the Africa diaspora community, mainly associated with the University of Cape Town, towards the modeling of a diaspora pension fund as a likely instrument for the financing of the continent's development infrastructure. The case study further sought to test, using chi-square and logistic regression, whether the independent variables of age, gender and education have an effect on the willingness/support of the members of the diaspora in the establishment of the diaspora pension fund for the financing of the continent's infrastructure. The results show that members of the sampled diaspora community support the modeling and also showed their willingness to be part of the diaspora pension fund and to have part of their pension contributions invested for the financing of the development infrastructure. The resulting model shows that males, who are middle-aged and well-educated, are more likely to be in support of the modeling of the diaspora pension fund as a likely instrument for financing the continent's development infrastructure. The results show that when modeling the diaspora pension fund factors such as age, education, gender, remaining years towards retirement, governance, corruption, regulatory issues, and the rule of law and the infrastructure of investment portfolio diversification have to be taken into account as they affect the likelihood of support amongst the diaspora community. The research will help policy makers, global pension funds, governments in Africa, and the bigger community of the Africa diaspora in assessing the feasibility of commercialising the diaspora pension fund as an innovative financing instrument.

Probiotics have been defined as “live microorganisms which when administered in adequate amounts confer a health benefit on the host”. The beneficial effects of probiotics in the gut are well described and roles including immunomodulation and colonisation resistance have been documented. Recent reports suggest that topical use of probiotic bacteria may be an effective strategy to promote skin health or inhibit disease. Therefore, in this thesis the potential of probiotics to protect skin from pathogenic bacteria was assessed using primary keratinocytes as a model system, and the skin pathogen, Staphylococcus aureus. The ability of three probiotics, L. reuteri ATCC 55730, L. rhamnosus AC413 and L. salivarius UCC118 to inhibit the growth of S. aureus was tested using well-diffusion assays and spot on the lawn assays. All three probiotics inhibited the growth of S. aureus in well-diffusion assays, though this property was dependent on growth medium. Inhibition of S. aureus growth was principally via the production of organic acids rather than bacteriocin production. Next, to determine whether probiotics could protect keratinocytes, confluent normal human epidermal keratinocytes (NHEK) were infected with S. aureus (106 CFU/ml) in the presence or absence of the probiotic (108 CFU/ml). NHEK viability was measured using trypan blue exclusion assays. L. reuteri had a significant protective effect on NHEK when applied 1h prior to (P=0.0003), or simultaneously with S. aureus (P=0.002). L. reuteri did not however protect NHEK when applied 1h after S. aureus addition. There was no change in the number of viable S. aureus in cell culture assays. To determine whether the protective effect was due to the inhibition of adhesion, NHEK were either pre-exposed to the probiotic for 1h, simultaneously exposed to the probiotic and S. aureus for 1h, or exposed to the probiotic 30 minutes after S. aureus addition for 1h. Pre-exposure of NHEK to L. reuteri (exclusion) and simultaneous exposure to L. reuteri and S. aureus (competition) resulted in significantly less staphylococci adhering to NHEK (P=0.03 and P=0.008 respectively). However when L. reuteri was added after S. aureus (displacement), the number of adherent staphylococci was not reduced. The necessity of S. aureus adherence for the inactivation of NHEK was demonstrated using a α5β1 integrin blocking antibody. Finally, to compare the innate response of NHEK to probiotics with S. capitis and S. aureus, TLR-2, antimicrobial peptide (AMP) expression and IL-8 production were measured. TLR-2 protein (but not mRNA) expression was reduced in the presence of S. aureus (P=0.018). NHEK pre-exposed to S. capitis prior to S. aureus infection however, exhibited elevated TLR-2 protein and mRNA expression (P<0.0001 and P=0.009 respectively). NHEK pre-exposed to L. reuteri prior to S. aureus had no significant change in TLR-2 expression compared to untreated controls. ELISAs demonstrated that IL-8 production was significantly increased in NHEK pre-exposed to L. reuteri prior to S. aureus infection (P=0.0001). In conclusion, L. reuteri protected NHEK from the toxic effects of S. aureus at least partly through competitive exclusion of binding sites on NHEK. Finally, NHEK innate responses to probiotic bacteria were akin to those to the skin commensal, S. capitis. L. reuteri induced expression of a neutrophil chemoattractant, suggesting it could be of importance in priming the innate immune response against S. aureus infections. Taken together, these results suggest that probiotic bacteria could be used prophylactically within skin creams and soaps to prevent S. aureus colonisation and infection in skin.

Made available in DSpace on 2015-04-14T13:36:05Z (GMT). No. of bitstreams: 1 390396.pdf: 742166 bytes, checksum: ff6f453c7552409274aceb3384b4bfd9 (MD5) Previous issue date: 2007-03-08<br>Base te?rica: Atualmente, a avalia??o progn?stica do c?ncer de mama inicial ? principalmente baseada nas caracter?sticas cl?nicas das pacientes e histol?gicas dos tumores. Recentemente, subtipos moleculares de carcinoma invasor da mama foram reconhecidos atrav?s dos estudos de perfilamento gen?tico por DNA microarray. Essa nova classifica??o pode melhorar a avalia??o progn?stica, mas essa nova tecnologia ainda n?o ? amplamente dispon?vel. Assim, a possibilidade de identificar os subtipos moleculares atrav?s de m?todos mais simples e baratos ? promissora. A identifica??o do subtipo basal-like de c?ncer de mama particularmente importante porque tem uma evolu??o desfavor?vel e menos op??es de tratamento dispon?veis. Adicionalmente, a express?o de Receptor para o Fator de Crescimento Epid?rmico (EGFR) parece ser mais freq?ente nesse subtipo. M?todos: Uma coorte retrospectiva de 112 pacientes consecutivas com carcinoma prim?rio de mama de est?gio patol?gico I ou II, diagnosticadas e tratadas na mesma institui??o entre 1995 e 2000, foi estudada. As caracter?sticas histol?gicas e cl?nicas, bem como dados de evolu??o cl?nica e sobrevida, foram revisados. An?lise imunohistoqu?mica foi realizada em blocos representativos dos tumores com anticorpos contra Receptor de Estr?geno (RE), Receptor para o Fator de Crescimento Epid?rmico Humano-tipo 2 (HER2), CK 5/6 e EGFR. O objetivo prim?rio foi identificar a preval?ncia do subtipo basal-like de c?ncer de mama (RE HER2-negativo e CK 5/6 e/ou EGFR positivo) nessa popula??o. Resultados: 13 dos 112 (11,6%) tumores avaliados foram do subtipo basal-like. Sua idade m?dia foi 49 anos; 77 % eram est?gio patol?gico II e 100 % eram carcinomas ductais invasores. N?o houve diferen?a progn?stica entre o subtipo basal-like e os subtipos luminal (RE positivo e HER2 negativo); com superexpress?o de HER2 (HER2 positivo) e indeterminado (quatro marcadores negativos) com rela??o ? sobrevida livre de doen?a e sobrevida geral. Esse resultado provavelmente se deve ao n?mero limitado de pacientes neste estudo e ao bom progn?stico de pacientes com c?ncer de mama em est?gio inicial. 11 dos 112 (10%) casos foram positivos para EGFR. Tumores positivos para EGFR foram mais freq?entemente negativos para RE (23 x 5%;p=0,01). Conclus?es: Nossos resultados sugerem que ? poss?vel identificar o subtipo basal-like de c?ncer de mama atrav?s da an?lise imunohistoqu?mica do RE, HER2, CK 5/6 e EGFR. Al?m disso, a express?o de EGFR parece ser mais freq?ente em tumores Re-negativos, o que sugere que drogas direcionadas contra o EGFR poderiam beneficiar esse subgrupo de pacientes

Thesis (Honors)--Ohio State University, 2005.<br>Title from first page of PDF file. Document formattted into pages: contains, 28 p. Includes bibliographical references (p. 27-28). Available online via Ohio State University's Knowledge Bank.

The portfolio has three parts. Part one is a systematic literature review, in which the empirical literature relating to the relationship between mental imagery and emotion is reviewed. Part two is an empirical paper which explores self-defining memories, mental imagery and emotion in individuals likely to develop bipolar disorder. Part three comprises the appendices.

Recent election campaigns generated extensive attention for their creative use of Internet, from President Obama’s 2008 Facebook tactic of allowing Facebook friends to share their support with each other to tools that allow supporters to mobilize and influence offline. This thesis asks whether Internet access alone can influence an individual to vote, within the context of American Presidential elections. First, I replicate similar literature by conducting a series of Linear Probability Models that indicate that Internet does have a significant impact on an individual’s decision to vote. However, one major issue that previous studies fail to address is the likelihood of endogeneity between self-reported Internet access and voting behavior. To address this, I introduce a measure of Internet Service Providers available in a given Congressional District as an instrumental variable. Once instrumented, it appears that Internet is largely insignificant. However, there is a key exception in 2008, where Internet access is significant. I argue that this is due to the developments in social media technology that revolutionized the ways in which candidates engaged with voters and voters engaged with one another.

<p> Gas hydrates, specifically methane hydrates, are sparsely sampled on a global scale, and their accumulation is difficult to predict geospatially. Several attempts have been made at estimating global inventories, and to some extent geospatial distribution, using geospatial extrapolations guided with geophysical and geochemical methods. The objective is to quantitatively predict seafloor total organic carbon and subsequently the geospatial likelihood of encountering methane hydrates. Predictions of TOC are produced using a sparsely observed dataset (Seiter et al., 2004) through a k-nearest neighbor (KNN) algorithm using 423 predictors and 7 nearest neighbors. KNN is unsupervised and non-parametric, as I do not provide any interpretive influence on prior probability distribution, so results are strictly data-driven. This TOC prediction, along with other global datasets (seafloor temperature, pressure, sediment thickness, and crustal heat flow) are used as parameters to train a KNN algorithm in identifying likely locations of methane and/or methane hydrate accumulation. I have selected as test sites several locations where gas hydrates have been well studied, each with significantly different geologic settings. These are: The Blake Ridge (U.S. East Coast), Hydrate Ridge (U.S. West Coast), and the Gulf of Mexico. I then use KNN to quantify similarities between these sites via the normalized distance in parameter space. Results on identification of likely methane and/or methane hydrate accumulation indicate the use of KNN as an unreliable method of identifying accumulation. However, global seafloor TOC predictions are reasonably accurate and have been incorporated to provide a potential analysis on hydrocarbon accumulation.</p><p>

A search is presented for vector-like quarks using 20.3 fb⁻¹ of proton-proton collisions at a center-of-mass energy of √S = 8 TeV collected with the ATLAS detector in 2012 at the CERN Large Hadron Collider. Vector-like quarks are predicted to exist in many theories beyond the Standard Model of particle physics that attempt to resolve the hierarchy problem. Events are selected containing jets including at least one b-jet, sizable missing transverse momentum, large scalar sum of the jet and lepton transverse momenta, and either three leptons or two leptons with the same electric charge. Standard Model processes rarely produce this final state and production of vector-like quarks would lead to an enhanced rate of such events. The data are interpreted in the context of a variety of models including pair production of vector-like quarks T and B, which have the same electric charge as the corresponding Standard Model quarks and can appear in either SU(2) weak isospin singlets, doublets or triplets. In addition single and pair production of the vector-like quark T₅/₃ which appears in an SU(2) weak isospin doublet or triplet and has electric charge 5/3, is considered. A moderate excess of data above the SM background expectation is observed with a significance of less than two standard deviations. The data are used to set limits at 95% Confidence Level (CL) on the new heavy quark mass for the various VLQ models considered using the CL_S method. The vector-like quarks T and B in the singlet model are excluded at 95% CL below a mass of 0.59 TeV and 0.62 TeV respectively. The T₅/₃ is excluded at 95% CL below a mass of 0.74 TeV when only pair production is considered and below 0.75 TeV when both pair and single production are considered.