Plant promoters are the principle cis-acting regulatory sequences responsible for the temporal and spatial expression of genes. One method for isolating plant promoters is based on the ability of a common soil bacterium, <i> Agrobacterium tumefaciens </i>, to transfer a specific segment of DNA (T-DNA) into plant cells. This specific T-DNA has been shown to integrate stably into the recipient plant genome. If the T-DNA contains a promoterless marker gene, then T-DNA integration events occurring adjacent and downstream to a promoter region can be detected by the activation of the marker gene. These T-DNA-mediated gene fusions, consisting of an unknown plant promoter sequence and the coding sequence of a marker gene, can be isolated using the marker gene as a promoter tag. The key objective of this work was to develop a novel, bifunctional selectable marker gene and assess its use as: a selectable marker gene in bacterial and plant transformation systems, and as a promoter tag for T-DNA promoter-tagging studies in dicots. A bifunctional fusion gene was produced between phosphinothricin acetyltransferase and neomycin phosphotransferase (PAT::NPT II), by fusing an NPT II coding sequence to the 3' terminus of the PAT gene. The PAT gene product confers tolerance to a non-selective herbicide L-phosphinothricin (Ignite, Hoechst AG). The neomycin phosphotransferase ('npt II') gene allows for direct selection of transformed cells with the antibiotic, kanamycin. Using an <i>in vivo Escherichia coli </i> selection system, a translational fusion gene between these two reporter genes was achieved. The resulting protein had activities of both parent enzymes. This was demonstrated both in transformed <i>Escherichia coli</i> and in transformed <i>Nicotiana tabacum</i> and <i>Brassica napus</i> plants. Using this bifunctional selectable marker gene, a T-DNA promoter tagging vector, pBAU2, was constructed and its utility was demonstrated in <i>Nicotiana tabacum</i>. One of the <i>N. tabacum</i> promoter tagged events was selected for subsequent promoter isolation studies. The promoter from this regenerant was isolated by screening a Lambda subgenomic library and also by thermal asymmetric interlaced (TAIL-)PCR. The isolated upstream regulatory sequence was fused to a reporter gene, â-glucuronidase ('gus'), and subjected to a preliminary evaluation in <i> Nicotiana tabacum</i> and in <i>Brassica napus</i>.

La coexistència de múltiples factors de risc cardiovascular com la síndrome metabòlica, l'obesitat abdominal, la dislipèmia i la resistència a la insulina, determinen l'aparició de la disfunció endotelial, primer event en la patogènia de l'aterosclerosi. La tonometria arterial perifèrica (PAT) és una tècnica novedosa i no invasiva la qual permet la valoració de la funció endotelial a nivell clínic. Per altra banda, evidències científiques recents suggereixen que FABP4, adipoquina secretada pel teixit adipós, circulant podria tenir un efecte directe en teixits perifèrics. Per tant, els nivells elevats de FABP4 podrien estar implicats en el dany arterial i la disfunció endotelial. Per dur a terme la valoració clínica es van reclutar pacients amb risc cardiovascular (CV) moderat i se'ls hi va realitzar el test de la funció endotelial per PAT i l'extracció sanguínia per l'anàlisi dels biomarcadors circulants endotelials, d'inflamació i oxidació lipídica i FABP4. A nivell in vitro, es va treballar en cèl·lules endotelials HUVECs i limfòcits (Jurkat T cells), i es va estudiar la via de senyalització de la PI3K/Akt/eNOS, l'expressió de molècules implicades en la disfunció endotelial i l'assaig d'adhesió dels leucòcits a l'endoteli. Els resultats més significatius a nivell clínic van ser que la sE-selectina (molècula d'adhesió endotelial) i les LDL oxidades s'associen inversament amb la mesura de RHI-PAT en els pacients amb risc CV moderat, és a dir amb una pitjor funció endotelial. La FABP4 plasmàtica es relaciona inversament amb el RHI-PAT en pacients amb diabetis tipus 2, per tant a més FABP4 més disfunció endotelial. A nivell in vitro, FABP4 podria ser capaç de modular les vies de senyalització de la insulina, disminuint les cascades de fosforilacions (Akt i eNOS) augmentant l'expressió de les molècules d'adhesió (VCAM1 i E-selectina) i augmentant l'adhesió de leucòcits a l'endoteli, induint la disfunción endotelial i la resistència a la insulina en les cèl·lules endotelials. Segons els resultats obtinguts, podem concloure que la sE-selectina i les LDL oxidades es relacionen amb el RHI-PAT en pacients amb risc CV recolzant el paper del RHI-PAT en la valoració clínica de la funció endotelial i els nivells circulants de FABP4 estan associats amb la disfunció endotelial en pacients diabètics. FABP4 podria induir la disfunció endotelial a nivell molecular, interferint en les cascades de senyalització depenents d'insulina en l'endoteli vascular. / The coexistence of multiple cardiovascular risk factors such as metabolic syndrome, abdominal obesity, dyslipidemia and insulin resistance, determine the onset of endothelial dysfunction, the first event in the pathogenesis of atherosclerosis. The peripheral arterial tonometry (PAT) is a novel and noninvasive technique which allows the assessment of endothelial function at the clinical level. Moreover, recent scientific evidence suggests that FABP4 circulating, adipokyne secreted by adipose tissue, could have a direct effect on peripheral tissues. Therefore, high levels of FABP4 might be involved in arterial damage and endothelial dysfunction. To perform the clinical assessment were recruited patients with moderate cardiovascular risk (CV). The endotehlial function was assessed by PAT. Antropometry, circulating endothelial, lipid oxidation and inflammation biomarkers and FABP4 were measured. In vitro, was working in endothelial cells HUVECs and leukocyte (Jurkat T cells), and were quantified protein expression and mRNA. The results were clinically significant in that the sE-selectin (endothelial adhesion molecule) and oxidized LDL are associated inversely with RHI-PAT in patients with moderate CV risk. The plasma FABP4 was inversely associated with RHI-PAT in patients with type 2 diabetes. In vitro, FABP4 might modulate insulin signaling, decreasing the phosphorylations pathways (Akt and eNOS) and increasing the expression of adhesion molecules (VCAM1 and E-selectin), inducing endothelial dysfunction and insulin resistance in endothelial cells. We conclude that sE-selectin and oxidized LDL are associated with RHI-PAT in patients with CV risk by supporting the role of PAT in the clinical assessment of endothelial function and circulating FABP4 levels are associated with dysfunction endothelial cells in diabetic patients. FABP4 might induce endothelial dysfunction at the molecular level, interfering in insulin-dependent signaling pathways in vascular endothelium.

The study's objective is to relate the development of Pat Metheny's stylistic characteristics from his interpretation of jazz standards to their incorporation into his own compositions. Stylistic elements are established and a sample of his compositions are analysed to compare his solo style in standards with his compositional style. Metheny is a recognised innovator in technique and uses a wide range of instruments in the guitar family, both traditional and radically new. The use of such instruments frees Metheny from some restrictions and the possibility that this freedom is a major influence in his improvisation and composition is remarked on. There is scope for further work based on a wider sampling, and the methodology used in this study could probably be modified to focus on this objective.

Este trabalho inova a aplicação de revestimentos duros usando o processo de soldagem SATG (Soldagem ao Arco Tungstênio com Atmosfera Gasosa) utilizando ligas, em forma de pó, incorporadas diretamente à poça de solda. Devido à boa resistência ao desgaste, utilizou-se a liga Stellite 6, no substrato SAE 1020. Dados de microdureza Vickers, taxa de deposição, diluição, análise micrográfica e desgaste foram comparados com resultados do PAT-P (Plasma por Arco Transferido-Pó) e permitiram avaliar a técnica proposta. Os resultados mostraram um revestimento pelo processo SATG-pó livre de poros, sem oxidação na superfície e diluição superior, ao processo PAT-P. Porém, o consumo de gás foi menor, devido à redução da vazão do gás de arraste e da não utilização de gás plasma. A análise microestrutural evidenciou uma microestrutura similar, para ambos os processos, mas o processo PAT-P apresentou uma microestrutura mais refinada. Obteve-se, pelo processo SATG-pó semi-automático, microdurezas entre 400 e 500 HV e uma taxa de diluição de 5%, próximas aos valores do processo PAT-P. Quanto ao desgaste, observaram-se valores também bem próximos para os três processos. / This work innovates the application of hardfacing by using the welding process GTAW (Gas Tungsten Arc Welding) using alloys in the powder form and directly embedded in the weld pool. Due to the good wear resistance it was used Stellite 6 alloy for hardfacing the substrate SAE 1020. Vickers hardness data, deposition rate, dilution, micrographic analysis and wear results were compared with the PTA-P (Plasma Transferred Arc-Powder) in order to evaluate the proposed technique. The results showed a coating produced by the process GTAW-powder free of pores, without surface oxidation and higher dilution in comparison with the PTA-P process. However, gas consumption was lower due to reduced flow of the carrier gas and as it was not used as plasma gas. Microstructural analysis showed a similar microstructure for both processes, but the PTA-P process presented a more refined microstructure. It was obtained, by the semi-automatic GTAW-powder process, micro hardness between 400 and 500HV and rate dilution of 5%, values close to the PTA-P process. As for the wear it was observed values very close too, for the three process.

Submitted by Geyciane Santos (geyciane_thamires@hotmail.com) on 2015-07-01T13:29:52Z No. of bitstreams: 2 Dissertação -Pilar Maria de Oliveira Moraes 1.pdf: 186319 bytes, checksum: b1332ee55824a51f80b4f32860e1af90 (MD5) Dissertação -Pilar Maria de Oliveira Moraes 2.pdf: 604576 bytes, checksum: 341618618f83b7aceabf5e7fc3c2d051 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-07T13:41:41Z (GMT) No. of bitstreams: 2 Dissertação -Pilar Maria de Oliveira Moraes 1.pdf: 186319 bytes, checksum: b1332ee55824a51f80b4f32860e1af90 (MD5) Dissertação -Pilar Maria de Oliveira Moraes 2.pdf: 604576 bytes, checksum: 341618618f83b7aceabf5e7fc3c2d051 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-07T13:39:34Z (GMT) No. of bitstreams: 2 Dissertação -Pilar Maria de Oliveira Moraes 1.pdf: 186319 bytes, checksum: b1332ee55824a51f80b4f32860e1af90 (MD5) Dissertação -Pilar Maria de Oliveira Moraes 2.pdf: 604576 bytes, checksum: 341618618f83b7aceabf5e7fc3c2d051 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-07T14:35:48Z (GMT) No. of bitstreams: 2 Dissertação -Pilar Maria de Oliveira Moraes 1.pdf: 186319 bytes, checksum: b1332ee55824a51f80b4f32860e1af90 (MD5) Dissertação -Pilar Maria de Oliveira Moraes 2.pdf: 604576 bytes, checksum: 341618618f83b7aceabf5e7fc3c2d051 (MD5) / Made available in DSpace on 2015-07-07T14:35:49Z (GMT). No. of bitstreams: 2 Dissertação -Pilar Maria de Oliveira Moraes 1.pdf: 186319 bytes, checksum: b1332ee55824a51f80b4f32860e1af90 (MD5) Dissertação -Pilar Maria de Oliveira Moraes 2.pdf: 604576 bytes, checksum: 341618618f83b7aceabf5e7fc3c2d051 (MD5) Previous issue date: 2010-12-14 / Outras / This study aims to identify cardiovascular risk factors and the impact of nutritional intervention in the industrial sector workers in Greater Metropolitan Belém, Pará, companies accredited to the Workers' Food Program (WFP). We studied 97 employees of the Experimental group (ExpG) and 90 control group (GCON), were included in the randomization workers showed that cardiovascular risk factors for elevated waist circumference (WC). Participated in the randomization of 18 workers and 20 ExpG GCON. For the randomization tests, we used the paired Student t test and Wilcoxon test. Anthropometric variables were identified as Body Mass Index (BMI) and WC, blood levels of cholesterol, triglycerides and glucose levels and socioeconomic characteristics, and used the chi-square test for evaluating the results. The analysis of food consumption was made by means of food frequency questionnaire using a methodology based on the level of consumption: <0.33. Food consumption low,> 0.33 and <0.66 average consumption of food;> 0.66 Food consumption high. It was identified a young population, most living with a partner, with family incomes from 1 to 2 minimum wages, having completed high school. Before the dietary intervention was observed anthropometric findings with a high prevalence of overweight and high levels of lipid fractions in GExp and GCON. The consumption of foods considered cardiovascular risk was high when compared to consumption of foods considered protective in GExp and GCON. The characteristics found related to lifestyle were the improvement in the practice of smoking revealed a low percentage of smokers and former smokers, moderate, moderate to high alcohol consumption and low frequency of physical activity in both groups. After randomization the GCON showed only one variable with significant difference and there is a decrease in systolic blood pressure. In the analysis of anthropometric findings were observed high prevalence of overweight and high levels of lipid fractions in both groups before the intervention and significant improvement of nutritional values in the intervention group nutrition. In the experimental group, there was a significant difference in three variables: the average decrease in BMI, HDL cholesterol increased, demonstrating an apparent improvement after intervention and LDL cholesterol and a decrease of the mean, ie, demonstrating beneficial effects of the action guideline the group of workers ExpG. There was a decrease in CC ExpG signaling a decrease in an important risk factor for developing cardiovascular disease does so with GCON. The action of nutritional intervention results showed significant improvements for the group that was submitted when associated with the control group, confirming that the nutrition education strategy proposed in the TSP, it has practical effects of positive health worker. / Este estudo tem por objetivo identificar os fatores de riscos cardiovasculares e o impacto da intervenção nutricional em trabalhadores do setor industrial na Região Metropolitana de Belém-Pará, de empresas credenciadas ao Programa de Alimentação do Trabalhador (PAT). Foram estudados 97 trabalhadores do Grupo Experimento (GEXP) e 90 do Grupo Controle (GCON), sendo incluídos na randomização trabalhadores que apresentaram fatores de risco cardiovascular elevados para Circunferência de Cintura(CC). Participaram da randomização 18 trabalhadores do GEXP e 20 do GCON. Para análises da randomização, foi utilizado o teste t de Student pareado e o teste de Wilcoxon. Foram identificadas variáveis antropométricas como Índice de Massa Corporal (IMC) e CC, níveis sanguíneos de colesterol, triglicerídeos e glicemia e características socioeconômicas, sendo usado o teste Qui-quadrado para avaliação dos resultados. A análise do consumo alimentar foi feita por meio de questionário de frequência alimentar utilizando a metodologia baseada no nível de consumo:< 0,33. Alimento de consumo baixo; >0,33 e <0,66 alimento de consumo médio; >0,66 Alimento de consumo elevado. Foi identificado uma população jovem, a maioria residindo com companheiro, com renda familiar de 1 a 2 salários mínimos, possuindo ensino médio completo. Antes da intervenção nutricional foi observado resultados antropométricos com alta prevalência de excesso de peso e índices elevados de frações lipídicas no GEXP e GCON. O consumo de alimentos considerados de risco cardiovascular foi elevado quando comparado ao consumo de alimentos considerados protetores no GEXP e GCON. As características encontradas relacionadas ao estilo de vida foram: a melhoria na prática do tabagismo revelando um percentual baixo de fumante e um moderado de ex-fumantes, moderado a alto consumo de álcool e baixa freqüência de atividade física nos dois grupos. Após a randomização o GCON apresentou apenas uma variável com diferença significativa, verificando-se diminuição da pressão arterial sistólica, podendo ser atribuída a Síndrome do Jaleco Branco, presença do profissional de saúde associada a elevação de pressão arterial. Na análise dos resultados antropométricos foram observados alta prevalência de excesso de peso e índices elevados de frações lipídicas nos dois grupos antes da intervenção nutricional. No grupo experimental, verificou-se a diferença significativa em três variáveis: diminuição da média do IMC, aumento do colesterol HDL, demonstrando aparente melhora após intervenção e o colesterol LDL, com a diminuição da média, ou seja, demonstrando efeitos benéficos da ação orientadora para o grupo de trabalhadores do GEXP. Houve diminuição CC no GEXP sinalizando uma diminuição de um fator de risco importante para desenvolvimento de doenças cardiovascular o mesmo não acontecendo com o GCON. A ação de intervenção nutricional apresentou resultados importantes de melhorias para o grupo a que foi submetido quando associado ao grupo controle, confirmando que a estratégia de educação nutricional proposta no PAT, tem efeitos práticos positivos para a saúde do trabalhador.

La qualité des produits commercialisés représente un enjeu essentiel dans le domaine pharmaceutique. De plus en plus, les industriels cherchent à contrôler les différentes étapes de leurs procédés, les produits générés à toutes les étapes de fabrication ainsi que leurs produits finis, le but de tout cela étant une meilleure compréhension des procédés, ainsi qu’un gain de temps et une réduction des pertes financières. La chaire de recherche sur les Technologies d’Analyse de Procédés (PAT) en partenariat avec Pfizer travaille sur le développement de connaissances sur les phénomènes physico-chimiques régissant le comportement des procédés, et sur les critères de transferts technologiques basés sur des modèles phénoménologiques ou mathématiques. C’est dans le cadre de cette chaire que s’est déroulée cette maîtrise. Les travaux présentés mettent en lumière le développement d’une méthode permettant d’analyser de manière non destructive 100% des comprimés fabriqués, à vitesse réelle de production. À l’aide de la spectroscopie Proche-Infrarouge, l’objectif est de quantifier la teneur en principe actif dans chaque produit et dans le but d’identifier tous ceux étant sur ou sous-dosés. Également, l’adaptation de cette méthode à d’autres types d’analyses qui ont un intérêt dans l’industrie, comme la quantification d’humidité des comprimés, et l’enrobage, a été étudiée. Dans chaque cas, des spectres proche-infrarouge de produits fabriqués ont été acquis à l’aide d’une sonde installée sur un système de tri fonctionnant à une vitesse supérieure à celle d’une presse (c’est à dire généralement de 60,000 à 160,000 comprimés par heure). Les données ont été utilisées pour bâtir des modèles multivariés et prédire les valeurs souhaitées. Les résultats obtenus ont montré qu’il était possible d’analyser l’intégralité des échantillons d’un lot, à une vitesse garantissant de ne pas ralentir la production. L’utilisation d’une telle méthode dans l’industrie permettra de contrôler la globalité des produits finis, et donc permettre une qualité de 100% des produits commercialisés et éviter les rappels. Elle permettra également d’être utilisée en développement, lorsque la génération de beaucoup de données est nécessaire, comme pour le développement d’un algorithme par exemple.

According to ICH Q8 process analytic technology (PAT) should be established to monitor critical quality attributes (CQAs) during manufacturing processes. Ethinylestradiol (EE) is a highly active and low dosed steroid hormone that is prone to oxidative degradation. The stability of this compound is therefore a critical quality attribute that has to be considered during drug formulation development. Beside the stability of EE, granule particle size and moisture are CQAs influencing the tableting ability of the resulting granules and the stability of EE. Both CQAs should be monitored during the production process. The investigations described in this thesis evaluate the implementation of in-line-sensors for monitoring of particle size (spatial filtering technology, SFT) and granule moisture (microwave resonance technology, MRT) during the fluid bed granulation process and the influence of process-variations on the stability of EE. The aim of these investigations was to develop an effective and mild fluid bed granulation process for a new model formulation based on microcrystalline cellulose as replacement for lactose as main filler excipient. The EE degradation products 6-alpha-hydroxy-EE, 6-beta-hydroxy-EE, 9(11)-dehydro-EE and 6-oxo-EE were quantified as an index for the stability of EE. It could be demonstrated that the surface of the filler substance influences the stability of EE due to the impact of water molecules. Hence, spraying sequence was determined to be a useful tool to improve the stability of EE. Correlations could be established for 6-oxo-EE with granule moisture and thermic parameters. The implementation of the SFT-sensor in the granulation process was successful. Measurement with the MRT-sensor for monitoring of granule moisture has to be improved.:Chapter I 1 1 Introduction Chapter II 41 2 Materials and Methods Chapter III 54 3 In-line Monitoring of Particle Size in a Fluid Bed Granulator: investigations concerning positioning and configuration of the sensor Chapter IV 72 4 Influence of in line monitored fluid bed granulation process parameters on the stability of Ethinylestradiol Chapter V 90 5 Influence of filler excipients on stability of EE Chapter VI 105 6 Discussion and Conclusion Chapter VII 131 7 Summary Chapter VIII 138 8 Zusammenfassung Reference List 146 Appendix 158

Purpose: This study assessed effectiveness of five tools recommended by the US Preventive Services Task Force (USPSTF), designed to help primary care clinicians determine which unaffected patients to refer to genetics specialists for breast cancer risk assessment based on concerning family history. Design: This descriptive secondary analysis included 85 women aged 40-74. All participants had a first-degree female relative previously diagnosed with breast cancer who also had uninformative negative BRCA1/2 tests. Methods: Each pedigree was evaluated using the five tools including the Family History Screen-7 (FHS-7), Pedigree Assessment Tool (PAT), Manchester Scoring System, Referral Screening Tool (RST), and Ontario-Family History Assessment Tool (Ontario-FHAT). All five tools were applied to each study participant. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated to describe each tool’s ability to identify women with elevated risk as calculated by the Claus model. Receiver operating curves (ROC) were also plotted. Differences between areas under the curve (AUCs) for all possible pairs of tools were estimated through logistic regression to assess for differences in tool performance. Results: Claus calculations identified 14 women out of 85 whose lifetime risk of breast cancer was elevated at > 15%. Only two tools, the Ontario-FHAT and FHS-7, identified all 14 women with elevated risk, a sensitivity of 100%. The FHS-7 tool flagged all 85 participants, meaning its specificity was zero. The Ontario-FHAT flagged 59 participants as needing referral (specificity 36.2%) and had a negative predictive value (NPV) of 100%, indicating that if a woman was not found to need a referral to a genetics professional, it is likely she did not have an elevated lifetime risk of developing breast cancer. AUC values were not significantly different between tools (all p values > .05), and thus were not helpful in discriminating between the tools. Conclusion: In this population, the Ontario-FHAT out-performed other tools in terms of sensitivity and negative predictive value; however, low specificity and positive predictive value must be balanced against these findings. Thus, the Ontario-FHAT can help determine which women would benefit from referral to a genetics specialist.

In 1991, British novelist Patricia Barker published Regeneration, the first of three novels that portrayed the exploits of both factual and fictional characters during the darkest days of WWI. Barker's Eye in the Door (1993), followed by The Ghost Road (1995) for which she won the Booker Prize for Fiction, completed the series that explored the effects of combat on the human psyche. What emerges as a dominant feature of Barker's war novels is her depiction of the ocular sense. Reminiscent of Orwellianism, Barker's texts contain a seemingly ubiquitous ocular presence. For example, neurasthenic patients are scrutinized by army psychiatrists, objectors and subversives are spied upon or imprisoned so that their activities may be observed, and combatants are faced with the challenge of reconciling the horrifying events they have witnessed in combat. This study investigates the role and importance of Pat Barker's depiction of eyes and visuality in her war trilogy. The overreaching goal of the thesis to examine Barker's aestheticized notion of ocularity. It is my aim to come some conclusions about how vision / ocularity signal the emergence of a few central themes in the texts such as power relationships, objectification, exposure and the transgression of boundaries. The social and linguistic theories of Michael Foucault, Roland Barthes, Georges Bataille, Martin Jay and others who have addressed the themes of perception and ocular symbolism will be introduced into my discussion with the aim of providing a theoretic foundation to many of my assertions. Chapters will begin with an interpretation of a piece of theoretical writing by one of these authors followed by an analysis of Barker's texts that incorporates the major tenets of that theory. These tenets will serve as a basis to my discussion and it is my hope that, through the creative application of theoretical writing, I will address a number of aspects of Barker's work, especially in relation to her ocular imagery, that that have thus far gone unexplored.

No description available.