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Orientador: Márcia Rossini Justino Mutton / Banca: Eduardo da Silva Martins / Banca: João Martins Pizauro Júnior / Resumo: A utilização de fermentação em estado sólido a partir de subprodutos agroindustriais possibilita a obtenção de várias biomoléculas de interesse industrial. As amilases são enzimas com grande aplicação nas indústrias têxtil, farmacêuticas, alimentícias e sucroalcooleira, representando aproximadamente 25% do mercado mundial. Neste trabalho foram isoladas três linhagens fúngicas (Rhizopus oryzae, Malbranchea pulchella e Chrysosporium zonatum), que apresentaram potencial amilolítico, quando cultivadas em três resíduos agroindustriais (quirera de arroz e de milho e farelo de trigo). Para a cepa que apresentou maior atividade enzimática, avaliou-se melhor substrato, tempo de cultivo, teores de umidade, fontes suplementares de nitrogênio, pH e temperatura de incubação, objetivando otimizar as condições de cultivo. Observou-se que a maior atividade enzimática foi obtida com a cepa de R. oryzae em 24 horas de fermentação, em meio de cultura contendo farelo de trigo como substrato, em temperatura de 35°C, utilizando solução de sais composta de NH4NO3 a 0,1% e MgSO4.7H2O a 0,1% como fonte de nitrogênio. Alterações de pH entre 4,0 e 6,0 não afetaram significativamente a síntese da enzima. A condições ótimas de atuação foram temperatura de 75°C e pH de 4,5. As enzima manteve-se estável a 75°C na ausência de substrato por 25 minutos / Abstract: The use of fermentation in a solid state from agroindustrial byproducts permits the obtaining of several biomolecules of interest the industry, like the enzymes. The amylases are enzymes with great application on the textile, pharmaceutical and food industries, representing around 25% of the worldwide market. In this work three ungal strains (Rhizopus oryzae, Malbranchea pulchella and Chrysosporium zonatum) were isolated. They presented amylolytic potential when cultivated in three agroindustrial by products (brewers rice, corn grits and wheat bran). To the strain that presented the greatest enzyme activity, it was evaluated the best substrate, cultivation time, moisture content, additional sources of nitrogen, pH and incubation temperature, aiming the optimization of cultivation conditions. I was observed that the greatest enzyme's activity was obtained with 24 hours of fermentation, in R. orizae in a culture medium that contained wheat bran as the substrate, at 35°C, using salt solution made with NH4NO3 0.1%, MgSO4.7h2O 0.1% and (NH4)2SO4 0.1% as nitrogen source. pH alterations between 4.0 and 6.0 didn't change significantly the enzyme's activity. The optimum conditions of performance of the enzyme were temperature of 75°C and pH 4.5. The enzyme kept stable in the lack of substrate for 25 minutes in 75°C / Mestre

Sociologists have been interested in how individuals in modern society are bound to each other since the inception of the discipline. The 'community question' has seen three broad paradigms in this time. The 'community lost' perspective argues that community belonging and modernity are incompatible. In contrast, 'community saved' research suggest that the nature of modernity has made the benefits of community more salient than ever. Finally, the 'community transformed' approach posits that community has metamorphosed to become more relevant to the modern world. This thesis seeks to explore the form that communities may take in contemporary urban settings, and investigate the processes that underlie their continuation. To this end, an ethnographic case study approach was employed to examine an organisation that appears to both offer and exploit community – Alpha. Alpha is an eleven-week catechetical course that seeks to integrate non-Christians into the Christian community. Identifying and employing three 'ideal type' categories of Alpha guests, I examine the methods that the organisation uses in its efforts to attract new members, keep them attending, and integrate them into the community of the church. I argue that Alpha offers three distinct forms of community, with each appealing to different ideal type guests. The first is a gateway to the larger community of the local congregation. The second is a low-commitment community-in-itself. Finally, Alpha represents a community that offers material benefits to members. Alpha suggests that communities can be successful in attracting members by creating hybridised, multifaceted forms of belonging but that fostering long-term commitment is more problematic. This supports the 'community transformed' position that argues that the forms of belonging that were dominant in pre- and early modernity are less salient today.

Orientador: Maria de Fátima Sonati / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-16T20:41:19Z (GMT). No. of bitstreams: 1 Suemasu_CintiaNatsumi_M.pdf: 6481597 bytes, checksum: f9093ca98b4efb60bfe4f81a7e80efee (MD5) Previous issue date: 2010 / Resumo: A talassemia alfa (a) constitui um grupo de doenças hereditárias causadas pela redução ou ausência da síntese das cadeias 'alfa' da hemoglobina. Os genes responsáveis pela codificação dessas cadeias são duplicados ('alfa' 2 e 'alfa'1) e estão localizados em um agrupamento gênico ou cluster, situado próximo ao telômero do cromossomo 16 (16p13.3). As deleções são as principais causas da doença, podendo afetar um ou ambos os genes 'alfa' do cromossomo (formas 'alfa' + e 'alfa' 0, respectivamente). Nos últimos anos, várias técnicas foram desenvolvidas para identificar as deleções envolvendo o cluster dos genes a no cromossomo 16. A gap- PCR, o Southern blot e o FISH são comumente aplicados com esta finalidade; entretanto, muitas deleções ainda não são detectadas utilizando-se essas estratégias metodológicas. Em 2005, uma técnica simples e adequada a análises quantitativas rápidas, o Multiplex Ligation-dependent Probe Amplification (MLPA), foi adaptada ao diagnóstico das hemoglobinopatias. Esta técnica é baseada na ligação e na amplificação, em reação de PCR, de dois oligonucleotídeos (sondas) que se hibridizam de forma adjacente ao DNA alvo. Cada oligonucleotídeo é projetado para gerar um produto de comprimento único e, por possuírem terminações comuns, todas as sondas podem ser amplificadas utilizando um único par de iniciadores. Por fim, o uso de fluorocromos permite a separação dos fragmentos, que foram gerados pela amplificação das sondas, em sistema de seqüenciamento por capilaridade. No presente estudo nós utilizamos o MLPA para investigar as bases moleculares da talassemia alfa em nove pacientes não relacionados, cinco deles com Doença da Hb H, cujos diagnósticos moleculares não puderam ser esclarecidos pelas técnicas usuais, incluindo o seqüenciamento de DNA. No total, foram utilizadas 44 sondas, distribuídas desde o telômero ao gene MSLN, cobrindo uma região de cerca de 800 kb, na região 16p13.3. Oito diferentes deleções foram detectadas. Enquanto quatro delas comprometem uma região de grande extensão que inclui todo o locus a e seu elemento regulatório, com tamanhos mínimos de 240 kb, 470 kb, 500 kb e 720 kb, respectivamente (posições 97000-334571, 97000-570532, 97000-602492 and 97000- 816477 do UCSC Genome Browser, Fevereiro, 2009, respectivamente), quatro outras encontram-se limitadas às regiões que contém o elemento regulatório, deixando os genes a intactos, porém sem expressão. Essas últimas apresentam tamanhos mínimos de 0.4 kb, em um dos casos, e de 95 kb a 100 kb nos demais (posições entre 163464-163904, 97000- 193847, 97000-202417 e 97000-202417 do UCSC Genome Browser, Fevereiro, 2009, respectivamente). Em um dos pacientes com fenótipo talassêmico heterozigótico foi demonstrada a presença de uma quadruplicação de cerca de 230 kb de DNA, incluindo todo o cluster a e seu elemento regulatório (97000 a 231370 do UCSC Genome Browser, Fevereiro, 2009). Este estudo representa o primeiro no Brasil a empregar o método de MLPA na caracterização das bases moleculares da talassemia a. A ampla diversidade de alterações identificadas enfatiza a necessidade de se investigar todos os casos cujos valores hematológicos revelem hipocromia e microcitose, na ausência de deficiência de ferro e níveis normais de HbA2 / Abstract: Alpha (a) thalassemia is an inherited hemoglobin disorder characterized by reduction or absence of the a-globin chain synthesis. These chains are encoded by duplicated genes ('alpha' 2 and 'alpha' 1) that lie in a gene cluster near the telomeric end of the short arm of chromosome 16 (16p13.3). Deletions are the major molecular cause of the disease and may affect one or both 'alfa genes in the chromosome (the 'alpha' + and 'alpha' 0 forms, respectively). In recent years several techniques have been developed to identify deletions involving the 'alpha' -globin cluster in chromosome 16. The molecular tests commonly used to identify these deletions are gap- PCR, Southern Blot analysis and FISH. However, several thalassemia deletions remain uncharacterized by these methods. In 2005 a simple technique suitable for rapid quantitative analysis ¾ multiplex ligationdependent probe amplification (MLPA) ¾ was adapted for the diagnosis of thalassemias. The approach is based on the ligation and PCR amplification of two adjacently hybridizing oligonucleotides (probes). Each oligonucleotide pair is designed to give a product of unique length, and by using common ends all the probes can be amplified with one primer pair. Finally, the use of a fluorescent label allows the probe to be separated in a capillary sequencing system. In this study we used MLPA to investigate the molecular basis of thalassemia in nine unrelated patients (five with Hb H disease) in whom the molecular causes of the disease could not be detected by sequence analysis or other conventional techniques. In total, 44 probe pairs were used, covering approximately 800 kb from the telomere to the MSLN gene in the 16p13.3 region. Eight different deletions were detected. While four of them affect a large region including the entire ? -globin gene locus and its upstream regulatory element, spanning genomic regions of at least 240 kb, 470 kb, 500 kb and 720 kb (positions 97000-334571, 97000-570532, 97000-602492 and 97000-816477 of the UCSC Genome Browser, February, 2009, respectively). The other four deletions were limited to a region that contains the upstream regulatory element; the ? -globin genes, although intact, are therefore not expressed. These deletions span a region of at least 0.4 kb in one case and from 95 kb to 100 kb in the other cases (positions 163464-163904, 97000- 193847, 97000-202417 and 97000-202417 of the UCSC Genome Browser, February, 2009, respectively). One carrier who was suspected of having heterozygous ? -thalassemia showed a segmental quadruplication spanning about 230 kb and including the a cluster and upstream regulatory element (97000-231370 do UCSC Genome Browser, February, 2009). This study is the first in Brazil to use the MLPA method to characterize thalassemias. The variety of rearrangements identified highlights the need to investigate all cases presenting with microcytosis and hypochromia without iron deficiency or elevated Hb A2 levels / Mestrado / Ciencias Biomedicas / Mestre em Ciências Médicas

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L’alpha-foetoprotéine (AFP) est la protéine majoritaire du sérum fœtal de mammifère. C’est une glycoprotéine produite et sécrétée par l’endoderme viscéral du sac vitellin, les hépatocytes fœtaux et dans une moindre mesure, par l’intestin fœtal. Son profil d’expression est onco-fœtal :la synthèse de cette protéine chute fortement après la naissance mais peut reprendre en cas de régénération hépatique ou en cas de tumeurs diverses. Cette protéine est capable de fixer les oestrogènes et jouerait un rôle dans la différenciation sexuelle du cerveau femelle.<p>\ / Doctorat en sciences, Spécialisation biologie moléculaire / info:eu-repo/semantics/nonPublished

Glucagon-like peptide-1 (GLP-1) has received much attention as a novel diabetes therapeutic due to its pleotropic blood glucose-lowering effects, including enhancement of glucose-stimulated insulin secretion, inhibition of gastric emptying and glucagon secretion, and promotion of beta-cell survival and proliferation. GLP-1 is produced in the intestinal L-cell via processing of the proglucagon precursor by prohormone convertase (PC) 1/3. Proglucagon is also expressed in the pancreatic alpha-cell; however, in this tissue PC2 is typically expressed instead of PC1/3, resulting in differential cleavage of proglucagon to yield glucagon as the major product. We hypothesized that expression of PC1/3 in the alpha-cell would induce GLP-1 production in this tissue, and that this intervention would improve islet function and survival. Initial studies in alpha-cell lines demonstrate that adenoviral delivery of PC1/3 to alpha-cells increases GLP-1 production. By encapsulating and transplanting either PC1/3- or PC2-expressing alpha-cells, the following studies show that while PC2-expressing alpha-cells increase fasting blood glucose and impair glucose tolerance, PC1/3-expressing alpha-cells decrease fasting blood glucose and dramatically improve glucose tolerance in normal mice and in mouse models of diabetes. We further show that transplantation of PC1/3-expressing alpha-cells prevents streptozotocin (STZ)- induced hyperglycemia. We also found that PC1/3-expressing alpha-cells also improve cold-induced thermogenesis in db/db mice, demonstrating a previously unappreciated effect of one or more of the PC1/3-derived proglucagon products. Studies in isolated mouse islets demonstrate that adenoviral delivery of PC1/3 to isolated islets increases islet GLP-1 secretion and improves glucose-stimulated insulin secretion and islet survival. Experiments with diabetic mice show that these GLP-1-producing islets are better able to restore normoglycemia in recipient mice following islet transplantation. Taken together, these studies demonstrate that the alpha-cell can be induced to process proglucagon into PC1/3-derived products, and that this shift redirects the alpha-cell from a hyperglycemia-promoting fate to a blood glucose-lowering one. This research opens up avenues for further investigating the therapeutic potential of inducing islet GLP-1 production in isolated human islets and in vivo in diabetes patients, and may represent a novel way to intervene in the progressive loss of beta-cells that characterizes diabetes. / Medicine, Faculty of / Cellular and Physiological Sciences, Department of / Graduate

Over the past decade, advances in ethylene oligomerization have witnessed explosive growth of interest from both commercial and academic standpoint, with chromium metal invariably being the metal of preference. A common feature in this literature was the extended long debate regarding the mechanism, metal oxidation states responsible for selectivity and the role of the ligand. This thesis work embarked on the isolation and characterization of new active intermediates called “single component catalysts” (or self activating) to address two important questions: (1) how the catalyst precursors re-arrange upon activation and (2) the real oxidation state of the activated species. Four different ligands systems have been examined for this purpose. The first part is a study on the NPIIIN ligand which can be described as a dynamic and non-spectator ligand. Upon aluminum alkyl activation, a series of single component chromium catalysts for selective ethylene oligomerization and polymerization have been isolated, fully characterized and tested. New selective single component chromium(I) catalysts have also been isolated and tested positively for ethylene trimerization. The second part includes a new series of chromium complexes based on the NPVN ligand. This ligands enabled to obtain the first polymer-free extremely active catalytic system. In both NPN ligand systems, a new activation pathway was discovered by using vinyl Grignard reagent [(CH2=CH)MgCl] as activator and/or reducing agent. The third part explores new modified pyrrole-chromium complexes which were found to be highly active and selective ethylene trimerization catalysts. This part was a continuation of previous work from our lab to complete the mechanistic picture of this highly successful pyrrole-chromium catalyst independently commercialized by Phillips-Chevron and Mitsubishi. Interestingly upon aluminum alkyl treatment, the first example of a Schrock-type chromium ethylidene complex has been isolated and characterized and found to be a potent catalyst for selective ethylene trimerization. Finally, the other ligands introduced in this thesis are new systems called pyridine-SNS and Si-SNS that introduce some modification to the known commercial SNS catalyst (Sasol technology). The introduction of a pyridine ring or a silyl unit in the ligand scaffold has allowed to understand the mechanism of action of this remarkable system.

This thesis proposes a novel chroma keying method based on stereo images, which can be applied to post-process the alpha matte generated by any existing matting approach. Given a pair of stereo images, a new matting Laplacian matrix is constructed based on the affinities between matching pixels and their neighbors from two frames. Based on the new matting Laplacian matrix, a new cost function is also formulated to estimate alpha values of the reference image through the propagation between stereo images.