Dissertations / Theses on the topic 'CPF1'

CPF1 (centromere promoter factor 1) of Saccharomyces cerevisiae is a multifunctional protein. This multifunctionality may be associated with the different forms of the protein. To date, no clear function has been attributed to this protein. The general roles currently attributed to CPF1 are: maintenance of proper chromosome segregation or centromere function; methionine prototrophy and chromatin modulation. A series of recent papers have suggested the possibility of CPF1 being involved in protein-protein interactions. This thesis shows that CPF1 has an additional protein-protein interaction domain which is a coiled-coil. A truncation of CPF1 coding for only the C-terminal dimerization motif was cloned into a 17 expression vector and the protein, Dd, was produced in and purified from E. coli. Characterisation of the conformation of the C-terminal dimerization domain of CPF1 show the presence of a leucine zipper in Dd and that Dd forms a dimer under physiological conditions. Sequence alignment analysis of CPF1 and other bHLHZIP proteins identified a conserved serine residue in the basic domain of CPF1 and a conserved asparagine residue in the leucine zipper. Point mutations were introduced separately mutating these residues into either alanine or glutamate. The mutant alleles were cloned into yeast expression vectors. The cloned alleles were used to transform YAG93, a cpf1 deletion strain. Characterisation of the transformants obtained indicate the possibility of CPF1 being involved in the spindle assembly checkpoint control, possibly as a tethering protein. Finally, attempts were made to identify interacting partners of CPF1 and a purification scheme was designed to purify a putative CPF1 complex. Initially, a GST-CPF1 chimera was used to screen radioactively labelled crude yeast extracts for interacting partners of CPF1. This strategy identified five or six potential proteins which might form a multiprotein complex with CPFl. Attempts were then made to purify this putative complex from S. cerevisiae. The partially purified extracts were tested for CDEI activity, indicative of the presence of CPF1 in the samples and. tjhen analysed for protein content. The samples were also tested in a Holiday junction binding assay and in a histone acetyltransferase activity assay. In conclusion, CPF1 is a bHLHZIP protein that has a role in nutrient signalling and checkpoint control, possibly by acting as an intermediary protein that recruits essential factors to gene promoters or the centromere by protein-protein interactions.

Background: Higher global competition and more demanding customers force manufacturing companies to develop and adopt new collaborative strategies; the collaborative planning, forecasting and replenishment concept allows stronger supply chain cooperation, cost saving, improved efficiency and performance. Purpose: Exploratory, to study the criticality of factors that can affect the feasibility of a potential a CPFR implementation in Sweden, from the perspective of a manufacturer in a pre-CPFR implementation stage.Method: Qualitative research, using a multiple case method of two manufacturing firms operating in Sweden. Using content analysis, it revolves around studying factor criticality, all the while showing differences and commonalities in terms capabilities, and future feasibility of CPFR between the two case studies. Findings: High degree of interconnectivity between the factors; the critical success factors for Sweden are relationship and trust, goal alignment, KPI’s and costs, with very important factors such as cross-functional communication, top management support, and lower impacting factors such as IT infrastructure and change management. Relationship and trust, cross-functional communication and change management are developed factors that the Swedish manufacturing firms possess. Implications: The study’s contributions are related to the criticality of factors that can affect CPFR implementations in Sweden’s manufacturing sector, showing the importance of each, contributing academically in attempting to fill in the gap related to CPFR in Sweden, and practically by allowing better strategic decision-making in relation to future implementations. This is even more relevant due to a thorough lack of research in this area.

This thesis intended to evidence the need for change in a SME to reduce levels of inventory based on their process related to their supply chain management. This research focused on CPFR which is a process innovation tool that stands for collaborative planning, forecasting and replenishment (Cassivi, 2006). The idea behind this process is to make collaborative actions on all members of the supply chain to come up with a share vision and objective. Based on the core concept of this process partners along the supply chain share information based on customer trends and needs to create a single forecast that is visualize at all times by its members to react accordingly to sudden changes in demand. The research´s main objective is to describe the main process needed for CPFR implementation in actual SME that is struggling in a supply chain that is under constant pressure and obtain insight on the benefits involved in this process to reduce levels of inventory. The research was made based on the concept of CPFR through the use of databases such as Google scholar to come up with a first idea of the concept and examples of other companies implementing CPFR as a their main process for their supply chain. The second approach was to contact the companies involved in our case and use questionnaires to identify the relationship and the problems involved in their supply chain structure. This methodology was used to establish a comparison between what has been done in other companies and what is needed in our case company. The companies involved in the research are a SME and a large manufacturer. The large manufacturer is working under CPFR but with final retailers setting aside the value of incorporating upstream suppliers. Due to bullwhip effect upstream suppliers are incrementing their warehousing facilities to react to changes on the demand. This is incrementing the cost of the operation and it is creating an unstable supply. The research demonstrated companies in where CPFR is proven to reduce stockouts, markdowns, levels of inventory, time to market, strengthen the relationship and overall reduce costs. Based on the discoveries in the retail industry it was clear the benefits from this process. Companies such as Condis revealed important information based on incorporating upstream suppliers. At the end observations were made based on implementation strategies such as following the 9 step guidelines set up by VICS, developing pilot programs, reducing the number of SKU´s at the beginning, working on seasonal products rather than standard products, and developing and implementing better IT systems to manage the level of information needed.

L’enzim Carnitina palmitoïltransferasa 1 (CPT1) permet l’entrada dels àcids grassos de cadena llarga a la matriu mitocondrial per tal de ser degradats i utilitzats així com a substrat energètic. El malonil-CoA, primer intermediari en la síntesi d’àcids grassos, és un inhibidor al•lostèric de CPT1, fet que permet una regulació coordinada entre la síntesi i l’oxidació d’àcids grassos en un mateix teixit, evitant així que es donin ambdós processos alhora. Una inhibició específica de CPT1, doncs, és una bona aproximació farmacològica per al tractament de desordres metabòlics que impliquin acumulació d’àcids grassos i resistència a insulina, com la diabetis tipus 2 i l’obesitat. La present tesi doctoral s’ha centrat en l’estudi de la regulació per malonil-CoA dels isotips hepàtic (CPT1A) i muscular (CPT1B) de CPT1, així com en l’estudi de l’isotip més recentment descrit de la proteïna d’expressió en cervell, CPT1C. Per tal d’aprofundir en l’estudi de determinants moleculars de CPT1 que marquen el seu grau d’inhibició per malonil-CoA, s’han realitzat construccions mutants i delecionades dels enzims CPT1A i CPT1B i s’han expressat en el llevat “Pichia pastoris”. Mitjançant l’anàlisi del paràmetre IC50, hem demostrat que la regió aminoterminal de l’enzim CPT1A de rata (residus 1-18) exerceix un efecte dominant sobre les posicions Glu590 i Met593 en determinar la sensibilitat a la inhibició per malonil-CoA. Paral•lament, s’han generat construccions amb la seqüència de l’enzim CPT1 de porc, que presenta una gran diferència en el seu comportament enzimàtic quan es compara amb l’enzim humà, tant l’isotip hepàtic com el muscular. En aquest treball s’ha identificat l’existència d’un determinant negatiu en la posició Glu17 que explica parcialment la menor sensibilitat d’aquest enzim a la inhibició pel malonil-CoA. A més a més, s’ha construït un model 3-D in silico de la CPT1B humana, que permet justificar observacions experimentals mostrades en aquest treball, tot i que no explica encara les diferències en el comportament cinètic entre els isotips hepàtics i musculars de la proteïna. CPT1C és l’isotip descrit més recentment, i encara avui se’n desconeix la seva funció, existeix controvèrsia respecte a la seva distribució subcel•lular i no se n’han descrit els mecanismes que en regulen l’expressió. Els resultats mostrats en aquest treball demostren que CPT1C és una proteïna sense activitat enzimàtica, independentment de la característica extensió C-terminal que presenta respecte els altres isotips. D’altra banda, el patró d’expressió observat del gen Cpt1c no és compatible amb el paper que se li ha atorgat sobre la regulació de la ingesta. A més a més, hem identificat l’expressió d’una isoforma soluble de CPT1C en cervell humà adult, que ofereix una eina útil per a l’obtenció d’un cristall de la regió C-terminal citosòlica de CPT1C.
STRUCTURE-FUNCTION RELATIONSHIP OF CPT1 PROTEINS CPT1 (Carnitine palmitoyltransferase 1) enables the entry of long chain fatty acids into the mitochondrial matrix, in order to be degraded and used as energetic substrate. Malonyl-CoA, the first intermediate in the synthesis of fatty acids, is an allosteric inhibitor of CPT1. Through CPT1 inhibition, it establishes a coordinated regulation between fatty acid synthesis and oxidation, thus preventing that both pathways coexist at the same time. A specific inhibition of CPT1 is therefore a good approximation or the pharmacological treatment of metabolic disorders involving accumulation of fatty acids and nsulin resistance, such as type-2 diabetes and obesity. This thesis has focused on the study of malonyl-CoA regulation of both CPT1A and CPT1B isotypes. In addition, we studied the more recently described isotype of the protein, CPT1C. To gain insight into the study of CPT1 molecular determinants of malonyl-CoA inhibition, mutants of both CPT1A and CPT1B enzymes were expressed in yeast Pichia pastoris. Analysis of the IC50 parameter, demonstrated that the aminoterminal region (residues 1-18) of the rat CPT1A enzyme has a dominant effect over Glu590 and Met593 positions in determining its sensitivity to malonyl-CoA inhibition. We also identified the existence of a negative determinant within the sequence of pig CPT1A (position Glu17), which partially explains the low sensitivity of this enzyme to malonyl-CoA inhibition. In addition, we designed an in silico 3-D model of human CPT1B, which still does not explain the kinetic differences between liver and muscle isotypes of the protein, but justifies some of our experimental data. CPT1C is the more recently described isotype of the protein. Its function still remains unknown, and controversy exists regarding its subcellular distribution and the mechanisms that regulate its expression. Here we show that CPT1C is a protein without enzymatic activity, regardless of its characteristic C-terminal extension, compared to the other isotypes. Moreover, the observed gene expression pattern is not compatible with its given role on the regulation of food intake. In addition, we identified the expression of a soluble isoform of CPT1C in human adult brain, providing a useful tool for obtaining a crystalline structure of the cytosolic C-terminal region of CPT1.

A gestão da cadeia de suprimentos, suas complexidades e os desafios relacionados à interação entre os elos, que são formados geralmente por empresas de culturas e estruturas diferentes, trazem desafios quanto à interação desses elos. Algumas iniciativas de gestão da cadeia de suprimentos, com conceitos de colaboração são utilizadas, com propósito de oferecer maior interação ao longo da cadeia de suprimentos. Uma dessas iniciativas denominada CPFR, propõe uma concepção colaborativa nas atividades de planejamento, previsão e reposição, e vem sendo adotada entre varejos e fornecedores. Seus resultados, suas vantagens, suas barreiras e os processos para a sua implantação através de uma metodologia adotada pela VICS® são expostos, entretanto, pouco se tem evidenciado nestes trabalhos a relação quanto à maturidade necessária para implantar uma atividade colaborativa na cadeia de suprimentos varejista. A colaboração talvez exija esforços que podem ser constituídos além das questões estruturais de uma organização. Este trabalho tem como objetivo propor uma avaliação destes aspectos colaborativos na implantação do CPFR. Para isso, o tema CPFR e a concepção da colaboração na cadeia de suprimentos foram revisados e foram utilizados multi-casos de uso onde o CPFR foi implantado no varejo brasileiro. Os resultados obtidos por esses estudos de casos apontam maiores dificuldades aos aspectos da colaboração exigidos na implantação do CPFR ou durante ela, do que os aspectos estruturais ou tecnológicos. Assim, o entendimento desses aspectos colaborativos em uso com a metodologia baseada no modelo da VICS® ou qualquer outra empregada, evidencia que o entendimento dos aspectos colaborativos ao implantar o CPFR nas empresas de varejo é um importante apoio ao processo de implantação.
The management of the supply chain, its complexities and challenges related to the interaction between the links, which are usually formed by companies from different cultures and structures, bring challenges to the interaction of these links. Some initiatives in supply chain management with collaboration concepts are used, with the purpose of providing greater interaction along the supply chain. One such initiative called CPFR, we propose a design in collaborative planning, forecasting and replenishment, and has been adopted between retailers and suppliers. Their results, its advantages, its processes and barriers to its implementation through a methodology adopted by VICS® are exposed, however, little has been evidenced in these studies the relationship in terms of maturity required to deploy a collaborative activity in the retail supply chain. The collaboration may require efforts that can be made in addition to structural issues within an organization. This work aims to propose a collaborative evaluation of these aspects in the implementation of CPFR. For this, the theme and design CPFR collaboration in the supply chain have been reviewed and were used multi-use cases where CPFR has been deployed in Brazilian retail. The results of these case studies point to the most difficult aspects of collaboration required in the implementation of CPFR or during her than structural or technological aspects. Thus, the understanding of these aspects in use with the collaborative methodology based on the model of VICS® or any other employee, shows that the understanding of the collaborative aspects when deploying CPFR in retail businesses is an important support the deployment process.

The current study evaluated the reliability and validity of psychological flexibility self-report measures between typically developing under 18 and over 18 aged individuals. An independent samples t test with a correlational statistical analysis was conducted to evaluate the relationship between the CPFQ, CAMM, AFQY and AAQII. The results demonstrated that three out of four measures are reliable and valid in determining psychological flexibility between the two groups. This study adds to literature that is conducive in the analysis of measures that are designed for younger population to be utilized by practitioners with older populations reliably.

Die Zapfenphotorezeptoren (ZPR) sind verantwortlich für das Scharf- und Farbensehen. Ein totaler Funktionsverlust der ZPR, welcher beim Menschen zur Achromatopsie führt, hat eine Degeneration der ZPR zur Folge. Die pathologischen Vorgänge, die zur Degeneration der ZPR führen sind noch nicht restlos entschlüsselt worden. Zur Analyse der ZPR-Degeneration werden Tiermodelle genutzt, wobei bisher nur wenige Tiermodelle vorhanden sind. Ziel dieser Arbeit ist die Untersuchung und Charakterisierung von Mausmodellen, die Mutationen in funktionell bedeutsamen Genen der ZPR tragen. Hierbei handelt es sich entweder um ein weiteres potentielles Kandidatengen für ZPR-Dystrophien, das SLC24A2 Gen, oder um das PDE6C Gen, welches als ursächlich für Achromatopsie identifiziert wurde. Die Tiere der verschiedenen Mauslinien wurden anhand ihrer Ohrkennung genotypisiert und für die verschiedenen molekularbiologischen Verfahren euthanasiert. Von den Augen wurde entweder das gesamte Auge, das eye cup, oder die Netzhaut weiterverarbeitet. Es wurde eine Methode zur Anrei¬cherung der ZPR mit fluorescence activated cell sorting (FACS) erarbeitet. Des Weiteren kamen Techniken auf der DNA-, RNA- und Protein- Ebene (PCR, RT-PCR, Klonieren, Western blot) sowie histologische und immunhistologische Techniken zur Anwendung. Die Tiere wurden in vivo mithilfe von bildgebenden Verfahren (Scanning Laser Ophthalmology (SLO)) untersucht und funktionell durch die Elektroretinographie (ERG) charakterisiert. Für diese Studie wurde die Cone-GFP (die grünes fluoreszierendes Protein in den ZPR produziert) Mauslinie zur Entwicklung der ZPR-Anreicherung eingesetzt und analysiert. Die slc24a2ENU-Maus¬mutante wurde extern hergestellt, wobei durch eine ENU-induzierte Mutagenese eine Punktmutation im slc24a2-Gen, welches für den NCKX2 Austauscher kodiert, an Position 1722 (c.1722g>t) gene¬riert wurde. Als weiteres Mausmodell wird die spontan aufgetretene Mausmutante cpfl1 (cone photo¬receptor function loss 1) untersucht, die zwei unterschiedliche Mutationen im pde6c-Gen besitzt. Durch Kreuzen dieser Mauslinien untereinander und mit der Trα -/- Mauslinie (knock-out Mutation der Stäbchen Transducin α-Untereinheit) wurden die Doppelmutanten Linien slc24a2ENUxCone-GFP, slc24a2ENUxTrα-/- und cpfl1xCone-GFP hergestellt und untersucht. Bei Cone-GFP Mäusen konnten ZPR mittels FACS angereichert und gezielt untersucht werden. In der angereicherten ZPR-Population wurde bei RT-PCR Analysen zum einen nur eine Spleißvariante des NCKX2-Austauschers detektiert, während bei RT-PCR Analysen von ganzen Augen mehrere Spleißvarianten des NCKX2-Austauschers gefunden wurden. Zum Zweiten wurden weitere Mitglie¬der der NCKX-Familie in der angereicherten ZPR-Population (NCKX-1, 4) detektiert. Bei der slc24a2ENU-Mausmutante konnte die Mutation im slc24a2-Gen sowohl auf DNA- als auch auf RNA- Ebene nachgewiesen werden, jedoch nicht auf Proteinebene. Sowohl bei morphologischen (SLO und Histologie) als auch funktionellen (ERG) Untersuchungen wurden keine Unterschiede zwischen Wildtyp und Mutante in der Retina erkannt. Mithilfe der SLO wurde bei allen Tieren eine verlängerte Persistenz der arteria hyaloidea beobachtet. Das ERG war bei der slc24a2xTrα-/- Dop¬pelmutanten gegenüber der reinen Trα-/- ohne Veränderung. Bei den slc24a2ENUxCone GFP Mäusen und den reinen Cone-GFP Mäusen konnte im Alter von einem Jahr keine Amplitude mehr im ERG gemessen werden. Bei der SLO (bei beiden Doppelmutanten) wurden bei einigen Mäusen autofluo¬reszierende Punkte, so genannte „AF-dots“ beobachtet. Eine Ursache hierfür konnte nicht erbracht werden. Vergleichende genomische DNA-Untersuchungen von Intron 4 des pde6c Gens zwischen der cpfl1-Maus und dem Wildtyp C57Bl/6 zeigten bei der cpfl1-Maus eine im Vorfeld bereits auf RNA-Ebene beschriebene 116bp Insertion als Anteil einer 1522bp großen genomischen Insertion ist. Zusätzlich wurde eine 1bp Deletion im Exon 7 des pde6c Gens bei der cpfl1-Maus detektiert (c.1042delT). Beide Mutationen bedingen eine Verschiebung des Leserasters (frame shift), das wiederum zu einem verfrühten Stopcodon führt (p.E289fsX297 und p.L348fsX362). Vergleichende RT-PCR Untersu¬chungen der pde6c Transkripte zwischen der cpfl1-Maus mit ihrem Parentalstamm BALB/c erbrach¬ten, dass beide die 116bp Insertion tragen. Bei der ERG Untersuchung der cpfl1-Maus (im Alter von drei und sechs Wochen) konnte keine ZPR-Antwort registriert werden. Bei einer vergleichenden SLO-Verlaufsstudie (3., 4., 6. und 8. Lebenswoche) zwischen der Cone-GFP und der Doppelmutanten cpfl1xCone-GFP Maus zeigte sich, dass die Anzahl der ZPR zu Beginn der Studie gleich sind, im weiteren Verlauf bei der Doppelmutante immer stärker abnehmen und mit 8 Wochen nur noch im ventralen Bereich vorhanden sind. Es konnte in dieser Arbeit gezeigt werden, dass die Cone-GFP Maus ein potentiell sehr wichtiges Mausmodell werden kann, um weiterführende Grund-lagenforschung an ZPR zu ermöglichen. Zum einen können ZPR durch FACS angereichert werden und zum anderen ist eine eindeutige und einfache Darstellung der ZPR in vivo durch die SLO-Technik möglich. Jedoch ist es wichtig, den Untersuchungszeitpunkt innerhalb der ersten 2 Monate anzusetzen, da die ZPR der Cone-GFP Maus danach fortschreitend degenerieren. Aufgrund der RT-PCR Untersuchung der NCKX-Familie in der angereicherten ZPR-Population der Cone-GFP Maus konnten weitere Mitglieder der NCKX Austauscher Familie in den ZPR identifiziert werden, wodurch die Aussage, dass der NCKX2-Austauscher die einzige Möglichkeit darstellt, um Ca2+-Ionen aus dem Außensegment der ZPR zu schleusen, relativiert werden muss. Die Slc24a2ENU-Mausmutante zeigt keinen offensichtlichen degenerativen retinalen Phänotyp. Die einzige Besonderheit stellt die sporadisch auftretende persistierende Arteria hyaloidea dar. Der bei einigen doppelmutanten Tieren auftretende retinale Phänotyp, der „AF-dots“-Phänotyp, ist möglich¬erweise auf den Stammhintergrund (C3H) zurückzuführen. Weitere Untersuchungen sind zur Klärung dieser Frage notwendig. Die cpfl1-Maus ist das erste Modell für Achromatopsie, bei dem die Pathologie durch Mutationen im pde6c-Gen ausgelöst wird. Das cpfl1-Allel besitzt zwei unterschiedliche Mutationen, die beide zu einem verfrühten Abbruch der Proteinsynthese führen. Durch RNA-Untersuchungen konnte gezeigt werden, dass die cpfl1-Maus sich nur in der 1 bp Deletion von dem Parentalstamm BALB/c unterscheidet. Jedoch scheinen beide Mutationen einen Anteil an der Herausbildung des Phänotyps der cpfl1-Maus zu haben. Im Rahmen der vergleichenden Verlaufsstudie mit Doppelmutanten cpfl1xCone-GFP wurde als pathologischer Vorgang eine Degeneration und keine Aplasie der ZPR nachgewiesen, da im Alter von 3 Wochen noch eine normale Anzahl an ZPR vorhanden ist.

In der vorliegenden Arbeit wurde die Funktion der beiden cyanobakteriellen Phytochrome Cph1 und Cph2 untersucht. Dafür wurde zunächst das Wachstum von Mutanten mit einem inaktivierten cph1- bzw. cph2-Gen unter verschiedenen Lichtbedingungen analysiert. Das Wachstum aller Phytochrommutanten war unter Starklicht beeinträchtigt. Dahingegen wuchs die cph1-Mutante im FRL schlechter als der Wildtyp, während das Wachstum der cph2-Mutante im RL vermindert war. Eine cph1/cph2-Doppelmutante zeigte unter allen Lichtbedingungen eine Wachstumsreduktion, die der jeweiligen Einzelmutante ähnlich war. Die genaue Ursache für die Beeinträchtigung des Wachstums der Phytochrommutanten konnte nicht ermittelt werden. Die verschiedenen Aspekte der Photosynthese, wie Pigmentzusammensetzung, maximale Netto-Sauerstofffreisetzungsrate und 77K-Fluoreszenzemission, waren in den Phytochrommutanten nicht signifikant verändert. Bei Synechocystis sp. PCC 6803 konnte eine lichtgerichtete Bewegung beobachtet werden, wobei man aufgrund von Aktionsspektren der Motilität eine Funktion der Phytochrome oder phytochromähnlichen Proteine bei der Steuerung der phototaktischen Bewegung vermutet. Dem Cph1-Protein konnte hierfür keine Rolle zugeordnet werden. Dahingegen scheint der Photorezeptor Cph2 die lichtgerichtete Bewegung der Zellen in Richtung einer Blaulichtquelle zu inhibieren. Eine Interaktion mit einem klassischen Blaulicht-Rezeptor konnte ausgeschlossen werden.
The function of the cyanobacterial phytochromes Cph1 and Cph2 was investigated. At first, the growth of mutants with an inactivated cph1 or cph2 gene was analysed under different light conditions. The growth of all phytochrome mutants was affected under high-light conditions. However, the cph1 mutant grew slower than the wild type in far-red light, whilst the cph2 mutant revealed a reduced growth under red light conditions. A decreased growth of the cph1/cph2 double mutant was observed under all light conditions with a growth rate similar to the corresponding single mutant. The exact reason for the growth impairment of the phytochrome mutants could not be ascertained. Different aspects of photosynthesis (pigment composition, maximal net-oxygen evolution and 77K fluorescence emission) were not changed significantly in the phytochrome mutants. Synechocystis sp. PCC 6803 shows a movement towards a light source. Based on action spectra of motility phytochromes and phytochrome-like proteins are supposed to have a function in regulating the phototactic movement. An influence of the Cph1 protein in the phototactic movement was not demonstrated. Whereas, the Cph2 protein seems to be involved in the inhibition of the cell movement towards blue light. An interaction with a typical blue-light receptor was excluded.

Dans ce memoire, nous decrivons une approche originale de synthese d'arbres moleculaires utilisant une voie organometallique. En effet, la complexation des composes aromatiques par le greffon cpfe#+ permet conjointement l'activation de groupes methyles portes par le noyau aromatique (reaction de peralkylation), l'activation de fonctions soit halogenees (substitution nucleophile aromatique), soit ether aromatique (reaction de coupure de la fonction ether). Utilisant ces possibilites, nous avons developpe la synthese de noyaux polyfonctionnels (oh, i) de multiplicite 3, 6 ou 9 et d'un synthon phenol poly-fonctionnel de progression 3, qui peut etre obtenu soit complexe au greffon cpfe#+, soit purement organique apres photolyse. Ceci a permis la synthese de composes polymetalliques comportant jusqu'a neuf centres redox transferant leur electron au meme potentiel. De facon similaire, l'obtention du phenol para tri-allyle a permis la synthese d'arbres moleculaires comportant 9, 18 et 27 fonctions ethyleniques peripheriques grace a la progression 3 qu'il introduit a partir des noyaux poly-iodes. Ces travaux ont ouvert la voie a de nombreuses perspectives en cours de developpement dans notre groupe de recherche

PONTIFÍCIA UNIVERSIDADE CATÓLICA DO RIO DE JANEIRO
CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO
A prática do Planejamento, Previsão e Reabastecimento Colaborativo (em inglês, Collaborative Planning, Forecasting and Replenishment – CPFR) é um tema recente de integração de cadeias de suprimento que tem sido amplamente abordado na literatura acadêmica. O CPFR é considerado, para muitos pesquisadores, uma iniciativa mais completa e avançada que outros métodos de colaboração na cadeia, sendo, em alguns casos, uma extensão ou aprimoramento dos mesmos. O amadurecimento do tema é representado pela existência de revisões sistemáticas na literatura. Contudo, por ser o CPFR ainda um tema recente com acentuado crescimento de estudos na literatura, esta dissertação tem como objetivo realizar uma atualização destas revisões sistemáticas para verificar novos avanços no assunto, se lacunas ressaltadas já foram pelo menos parcialmente endereçadas e se tendências estão se consolidando. Mais ainda, em função da dificuldade de se implementar o CPFR na prática, um enfoque maior na análise dos estudos empíricos existentes com o objetivo de ir além de resultados parciais e particulares oferecidos na literatura é contemplado nesta dissertação, expandindo assim o escopo das revisões existentes com o foco específico em estudos empíricos. A dissertação apresenta seus resultados guiados em um framework de síntese para integração de cadeias focado em CPFR, tendo como base os passos para a condução de uma pesquisa empírica em gerência de operações, os blocos conceituais do CPFR e a contribuição dos estudos empíricos. Uma agenda para pesquisas futuras finaliza a dissertação.
The practice of Collaborative Planning, Forecasting and Replenishment (CPFR) is a recent topic of supply chain integration that has been extensively covered in academic literature. The CPFR is considered by several researchers a more complete and advanced initiative than other methods of collaboration in the chain, being in some cases an extension or improvement of them. The maturation of the theme is represented by the existence of systematic reviews in the literature. However, since CPFR is still a recent topic with a strong growth of studies in the literature, this dissertation aims to carry out an update of these systematic reviews in order to verify new advances in the subject, if highlighted gaps have been at least partially addressed, and also if trends have been consolidating. Moreover, due to the difficulty of implementing CPFR in practice, a greater focus on the analysis of existing empirical studies with the objective of going beyond partial and particular results offered in the literature is contemplated in this dissertation, thus expanding the scope of the existing reviews with specific focus on empirical studies. The dissertation presents its results guided by a synthesis framework for supply chain integration focused on CPFR, based on the steps for conducting an empirical research in operations management, the conceptual building blocks for CPFR and the contribution of the empirical studies. An agenda for future research concludes the dissertation.

Planejamento, Previsão e Suprimento Integrado (Collaborative Planning Forecasting and Replenisement- CPFR)é um tema que está evoluindo a nível mundial, onde aplicaçõespráticastêm indicadopotencialparapromovera economiae a melhoriano nível de serviço das empresas. O CPFR une a demanda final de produtos com a cadeia produtiva dos mesmos, criando, assim, um processo único em que todos os envolvidos são beneficiados através de relações colaborativas de parceria. A pesquisa apresenta uma revisão teórica sobre o tema, abordando os níveis de colaboração possíveis entre empresas, exemplos de aplicação do CPFR e os resultados obtidos, as diretrizes de implementação com ênfase nos nove passos propostos pela VICS (The Voluntary Interindustry Commerce Standards), os habilitadores necessários para implementação, as etapas envolvidas no desenvolvimento de um projeto piloto e as barreiras para uma implementação. Um estudo de caso é apresentado com uma proposta de aplicação da metodologia CPFR efetuada por uma empresa ligada à produção de insumos para um varejista atuante no mercado moveleiro, onde são destacados os indicadores críticos de performance do processo CPFR. As conclusões obtidas referem-se à aplicabilidade do modelo sugerido pela VICS no Brasil, para a obtenção de resultados semelhantes aos atingidos pelas empresas que já estão utilizando CPFR em seus processos colaborativos de relacionamento.

Orientadores: Sergio Valdir Bajay, Gilberto De Martino Jannuzzi
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Mecânica
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Resumo: Durante as últimas décadas, têm-se introduzido poucas modificações no setor elétrico brasileiro no que se refere às novas modalidades de tarifas para os consumidores finais. As mudanças mais radicais foram implantadas no início da década de oitenta com a introdução das tarifas horosazonais verde e azul. O objetivo desta dissertação é testar as possibilidades de se formular novas tarifas horosazonais no País, através de uma análise crítica da aplicação destas no Brasil e no mundo, e a realização de um estudo de caso envolvendo pesquisas de campo, entrevistas e medições em consumidores industriais atendidos na categoria tarifária A4. A análise da experiência nacional aborda a implantação e o desenvolvimento das tarifas horosazonais - verde, azul e amarela - e tarifas de fornecimento interruptível no País por meio de um histórico completo dos fatos mais importantes, ocorridos desde 1957 até 2006, e de uma avaliação dos resultados a que se chegou com a implantação destas tarifas. Na análise da experiência internacional, examinaram-se os casos de França, Canadá, Estados Unidos e Portugal. O estudo de caso envolveu três pesquisas de campo. A primeira delas teve como objetivo escolher os segmentos industriais com maior potencial para a modulação de carga. Com a segunda pesquisa, conseguiu-se conhecer melhor os consumidores dos segmentos de calçados e de móveis, em termos de perfil de demanda e de detalhes de seus processos produtivos, visando estabelecer, com mais segurança, suas possibilidades de modulação de carga. A terceira pesquisa de campo permitiu o levantamento dos dados técnicos e econômicos, necessários para se fazer simulações e uma avaliação quantitativa dos impactos econômicos decorrentes de um terceiro posto tarifário para estes consumidores, na madrugada. As análises custo/benefício, realizadas no estudo de caso, levaram em conta tanto a ótica do consumidor como a da concessionária. Os resultados das simulações realizadas mostraram que os custos com mão-de-obra chegam a ser 35 vezes maiores do que aqueles com a fatura de eletricidade nestes segmentos industriais. Isto desestimula um possível deslocamento de parte da produção para o período noturno, por conta do adicional noturno no custo da mão-de-obra, mesmo com tarifas de energia elétrica muito baixas neste período
Abstract: During the last decades, the Brazilian electric sector has introduced little changes in tariffs for end-use consumers. The most radical changes were implanted at the beginning of the eighties, with the introduction of a seasonal tariff structure called green and blue. The objective of this work is to test some possibilities to formulate a new seasonal tariff in Brazil, through a critical analysis of these tariffs in Brazil and in other countries. This was made through field researches, interviews and the measuring of some industrial consumers. The national experience analysis includes the implementation and development of the seasonal tariffs - green, blue and yellow -, the supply curtailable rate and a complete historical of the more important facts occurred since 1957 up to 2006, together with an evaluation of the impacts in the implementation of these tariffs. In the analysis of the international experience, it was examined tariffs in France, Canada, United States and Portugal. The case study involved three researches on the field. The first one was to choose the industrial segments with larger potential for load modulation. The second research explored both footwear and furniture industries in terms of theirs demand profile and productive processes, with the objective of establishing their load modulation change possibilities more accurately. The third field research obtained the necessary technical and economical data, to work with simulations in a quantitative analysis of the economic impacts of a third tariff position during night time. The cost-benefit analysis considered both consumers and Utility's point of views. The results of the simulations has shown that the cost of labor is sometimes 35 times higher than electricity bills in the footwear and furniture industries, rendering useless a possible load modulation change during the night. Furthermore, during night time the cost of labor increases
Mestrado
Planejamento de Sistemas Energeticos
Mestre em Planejamento de Sistemas Energéticos

Pressurized chemical looping combustion (PCLC) is a promising next generation carbon capture technology which operates on the fundamentals of oxyfuel combustion to concentrate carbon dioxide in the flue gas stream. Oxygen is supplied through cyclic oxidation and reduction of a solid metal oxide between an air reactor and fuel reactor to prevent the direct contact of fuel and air. CanmetENERGY-Ottawa, in collaboration with Hatch Ltd., is designing a pilot scale PCLC system which uses ilmenite as the oxygen carrier and a novel fluidized bed design called the Plug Flow Internally-recirculating Reactor (PFIR). The PFIR consists of an annular bubbling fluidized region in which particles are circulated by angle jets through two reactive zones separated by baffles. The overall objective of this thesis was to provide key design parameters and insight for the construction of the pilot facility. Experimental work was first conducted investigating the minimum fluidization velocity (Umf), gas bubble size, and tube-to-bed heat transfer coefficients of different ilmenite particle size distributions (PSDs) at varying pressures up to 2000 kPa. The data was compared to a variety of literature correlations. The Saxena & Vogel (1977) constants for the Wen-Yu type correlations (Remf=√C12+C2Ar-C1) resulted in the best fit for predicting the Umf of the PSDs with Sauter mean diameters (SMD) less than 109 μm, while the Chitester et al. (1984) constants resulted in better predictions for the larger particle size distributions (SMD greater than 236 μm). Gas bubble size was found to be marginally impacted by pressure, with the Mori & Wen (1975) correlation best fitting the data. The heat transfer coefficient was found to also be marginally increased by pressure with the the Molerus et al. (1995) correlation matching the atmospheric data. A computational particle fluid dynamic (CPFD) model of the experimental unit was then created and validated using the obtained data for minimum fluidization velocity and bubble size. The accuracy of the model was found to be dependent on the particle close packing factor input variable, with a value of 0.58 resulting in the best results for each of the ilmenite PSDs modeled. Finally, a CPFD model was created for a cold flow design of the PFIR to investigate the impacts of different operating parameters on the solids circulation rate and gas infiltration rate between the two reactor zones. This model used the validated parameters of the previous CPFD model to add confidence to the results. The impacts of increasing superficial gas velocity, fluidizing gas jet velocity, bed height, and pressure were all found to increase the solids circulation rate through their respective impacts on the momentum rate of the fluidizing gas. A polynomial function was fit between these two variables resulting in a method to predict the solids circulation rate. Similarly, the rate of gas infiltration between sections was found to be dependent on the solids circulation rate, allowing for a function to be made to predict the gas infiltration at different operating conditions.

The accumulation of salts in soil is an important agricultural problem that limits crop productivity. Salts containing sodium (Na⁺) are particularly problematic, as cytosolic Na⁺ can interfere with cellular metabolism and lead to cell death. Maintaining low levels of cytosolic Na⁺, therefore, is critical for plant survival during growth in salt. Mechanisms to regulate Na⁺ accumulation in plant cells include extrusion of Na⁺ from the cell and sequestration of Na⁺ into intracellular compartments. Both of these processes are controlled in part through the action of Na⁺/H⁺ exchangers belonging to the Cation/Proton Antiporter-1 (CPA1) gene family. Genes belonging to this family have been identified in both salt-sensitive and salt-tolerant species, suggesting that salt-tolerant species may have evolved salt tolerance through modification of these existing pathways. The research presented here has focused on understanding how salt tolerance has evolved in Brassicaceae species, and particularly on the role that CPA1 genes have played in the adaptation to salinity of Eutrema salsugineum. Specific projects have sought to understand 1) how copy number variation and changes in coding sequences of CPA1 genes contribute to salt tolerance in E. salsugineum and its salt-tolerant relative Schrenkiella parvula, 2) whether functional or regulatory changes in Salt Overly Sensitive 1 (SOS1) from E. salsugineum (EsSOS1) contribute to its enhanced salt tolerance, and 3) whether accessions of Arabidopsis thaliana differ significantly in their response to salt stress.The results indicate that EsSOS1 and SOS1 from S. parvula (SpSOS1) both confer greater salt tolerance in yeast than SOS1 from A. thaliana (AtSOS1) when activated by the complex of the SOS2 kinase and SOS3 calcium-binding protein, whereas only EsSOS1 confers enhanced salt tolerance in the absence of activation. When expressed in A. thaliana, EsSOS1 also confers greater salt tolerance than AtSOS1 through regulatory changes that likely involve differences in expression pattern. Together, the results presented here suggest that mechanisms regulating cellular Na⁺ accumulation that exist in salt-sensitive crop species could be altered to enhance growth in salty soils. In addition, the 19 A. thaliana accessions used to create the MAGIC population were shown to differ significantly in their response to salt stress.

The protection of sensitive data and technologies is critical in preserving United States (U.S.) national security and minimizing economic losses. However, during a cyber attack, the operational capability to constrain the exfiltrations of sensitive data and technologies may not be available. A cyber preparedness methodology (CPM) can improve operational capability and cyber security. The CPM enables a corporation to (a) characterize cyber threats; (b) determine the level of preparedness necessary to ensure mission success; (c) facilitate strategic planning for cyber security (CS); and (d) establish priorities for CS investment planning and management decisions. The cyber preparedness framework (CPF) underlies the CPM. A corporation's leadership articulates its fundamental approach to risk management (RM) and mission assurance, and determines its target level of preparedness. Typically, corporations utilize the CPF to (a) characterize the caliber of the threat; (b) assess the technical and operational capabilities to counter the threat; and (c) develop the governance and processes necessary to achieve its cyber preparedness level. The problem that was investigated in this case study was how to construct a CPF for Lockheed Martin (LM) that works in conjunction with a risk management process (RMP). The goal was to extend the CPF into an RMP to construct a risk management framework (RMF) paradigm that can aid similarly large-sized private sector U.S. Government (USG) contractors in implementing the CPM. In this investigation, the author identified the corporate (a) security categorization, (b) cyber threats, (c) cyber threat level, (d) cyber preparedness level, (e) capabilities the corporation should utilize to counter cyber threats, and (f) governance and processes necessary to achieve the cyber preparedness level for a large-sized private sector USG contractor. The results of this investigation were organized in terms of RMP phases. Based on the results, the author constructed an RMF paradigm that can aid similarly large-sized USG contractors in implementing a CPM.

Molti metodi di compressione lossless si basano sulle idee che nel 1977 i ricercatori israeliani Abraham Lempel e Jacob Ziv hanno presentato nell’articolo “A universal Algorithm for sequential Data Compression”. In questa tesi viene descritto il metodo di fattorizzazione LZ77, illustrato appunto da Lempel e Ziv, e vengono esposte le strutture dati fondamentali per la sua realizzazione. Sono inoltre descritti due algoritmi CPS1 e CPS2 che realizzano LZ77. Infine, sfruttando i dati raccolti sperimentalmente da Al-Haffedh et al. in “A Comparison of Index-Based Lempel-Ziv LZ77 Factorization Algorithms” [2012], gli algoritmi descritti vengono confrontati in termini di spazio e tempo.

Historically, South African manufacturers and suppliers to the mass retail environment have been compelled to manage significant business risks as a result of the generic buying strategies employed by the mass retailing format. More recently, best practice initiatives such as SCOR’s collaborative planning, forecasting and replenishment have risen to the fore of supply chain management as ways to mitigate the undesired effects of theses generic buying practices for all participants in the value chain.Traditional thinking centred on optimising only the merchandise activities and function, through cost based performance measures, have caused a number of undesired effects and invalid assumptions. These factors in turn have impacted the competitiveness and sustainability of manufacturers and suppliers as well as the supply chain ecosystem as a whole. Systemic theory suggests that in order to identify these conflicting and invalid assumptions one must approach the problem through sufficiency based thinking processes that communicate the core conflict and map out possible solutions for managers. Data for this study was collected based on the widely accepted best practice framework of supply chain management for the mass retail environment. With this in mind, this research aims to provide an academic foundation for deeper collaboration between mass retailers and their vendors, as well as an understanding of the practical implications of decisions for managers and executives, on both the mass retail, and manufacturing and supply sides of the value chain.While statistical variation is a reality in the retailing environment, the mass retailing format and its supply chain partners are particularly susceptible to the negative effects of ‘bullwhip’ due to the large scale of promotional activities undertaken. Much of this problem can be mitigated through collaboration on a meaningful bases that allows not only for responsiveness for supply chain partners but greater profitability for all participant in the value chain. It is argued that an improvement in throughput will have a positive impact on the competitiveness and sustainability of the local supply and manufacturing organisations in South Africa.
Dissertation (MBA)--University of Pretoria, 2012.
Gordon Institute of Business Science (GIBS)
unrestricted

CP1 (encoded by the gene CEP1) is a sequence-specific DNA-binding protein of Saccharomyces cerevisiae that recognizes a sequence element (CDEI) found in both yeast centromeres and gene promoters. Strains lacking CP1 are viable but exhibit defects in growth, chromosome segregation, and methionine biosynthesis. To investigate the basis of the methionine requirement, a YEp24-based yeast genomic DNA library was screened for plasmids which suppressed the methionine auxotrophy of a cep1 null mutant. The suppressing plasmids contained either CEP1 or DNA derived from the PHO4 locus. PHO4 encodes a factor which positively regulates transcription of genes involved in phosphate metabolism via an interaction with CDEI-like elements within the promoters of these genes. Subcloning experiments confirmed that suppression correlated with increased dosage of PHO4. PHO4c, pho80, and pho84 mutations, all of which lead to constitutive activation of the PHO4 transcription factor, also suppressed cep1 methionine auxotrophy. The suppression appeared to be a direct effect of PHO4, not a secondary effect of PHO regulon derepression, and was dependent on a second transcriptional regulatory protein encoded by PHO2. Spontaneously arising extragenic suppressors of the cep1 methionine auxotrophy were also isolated; approximately one-third of the them were alleles of pho80. While PHO4 overexpression suppressed the methionine auxotrophy of a cep1 mutant, CEP1 overexpression failed to suppress the phenotype of a pho4 mutant; however, a cep1 null mutation suppressed the low-Pi growth deficiency of a pho84 mutant. The results suggest that CP1 functions as a transcriptional regulator of MET genes, and that activation of PHO4 restores expression to those genes transcriptionally-disabled by the cep1mutation. The results also suggest the existence of a network that cross-regulates transcription of genes involved in methionine biosynthesis and phosphate metabolism. A direct molecular approach to investigate CP1's role in MET gene expression was also taken. CDEI sites are associated with the promoter regions of most MET genes, but only MET16, the gene encoding PAPS reductase, has been shown to require CP1 for expression; both PAPS reductase activity, and MET16 mRNA are absent in cep1 mutants. Results of the present study demonstrate that CP1 participates in two systems which regulate expression of MET16, one triggered by methionine starvation and requiring the transactivator MET4 (pathway-specific control), and the other triggered by starvation for many different amino acids and requiring GCN4 (general control). CP1 was shown to mediate its regulatory function through the upstream CDEI site, and to act directly or indirectly to modulate the chromatin structure of the MET16 promoter. In addition, the pho80 mutation was found to partially restore MET16 expression to the cep1 strain, confirming the proposed nature of PHO4 suppression. A second methionine biosynthetic gene MET25, was also analyzed. Like MET16, MET25 was found to be regulated by both pathway-specific and general control mechanisms, but in contrast to MET16, CP1 only participated in the pathway-specific response of this gene. The results demonstrate that CP1, possibly by modulating changes in chromatin structure, assists the regulatory proteins MET4 and GCN4 in activating transcription of MET genes.

CP1 is a sequence specific DNA-binding protein of the yeast Saccharomyces cerevisiae which recognizes the highly conserved centromere DNA element I (CDEI) of yeast centromeres. The gene encoding CP1, which was designated CEP1 for centromere protein 1, was cloned and sequenced. CEP1 encodes a highly acidic protein of molecular weight 39,400. CEP1 was mapped to a position 4.6 centiMorgans centromere distal to SUP4 on the right arm of chromosome X. Phenotypic analysis of cep1 mutants demonstrated that yeast strains lacking CP1 are viable but have a 35% increase in cell doubling time, a ninefold increase in the rate of mitotic chromosome loss, and are methionine auxotrophs. Detailed analysis of the mitotic chromosome-loss phenotype showed that the loss is primarily due to chromosome nondisjunction (2:0 segregation). During meiosis cep1 null mutants exhibited aberrant segregation of centromere containing plasmids, chromosome fragments, and chromosomes. The predominant missegregation event observed was precocious sister segregation. The mutants also displayed a nonrandom 20% decrease in spore viability. Missegregation of chromosomes accounted for some but not all of this decreased spore viability, the remainder of which is presumed to be related to the pleiotropic consequences of the cep1 mutation. Together with the observed mitotic missegregation phenotype the results are interpreted as suggesting that CP1 promotes sister chromatid-kinetochore adhesion. The following conclusions are based on my mutational analysis of CP1: (1) CP1 is normally present in functional excess, (2) the C-terminal 143 amino acids are sufficient for full CP1 function in chromosome segregation and methionine metabolism, and (3) while DNA binding is apparently necessary for function, DNA binding per se is not sufficient. All of the mutations which caused an observable phenotype affected both centromere function and methionine metabolism. In addition, a direct correlation was observed in the degree to which both phenotypes were affected by different mutations. None of the mutant proteins displayed trans-dominant effects in a wild type background; however, two nonfunctional DNA binding-competent mutants exerted a dominant negative effect on the ability of PHO4 to suppress cep1 methionine auxotrophy. The data are consistent with a model in which CP1 performs a similar function at centromeres and promoters.

La β-oxidació dels àcids grassos al mitocondri és una de les principals fonts per a l’obtenció d’energia per a les cèl·lules. Per a que ocorri, cal que els àcids grassos siguin transportats a la matriu mitocondrial; aquesta funció és duta a terme per enzims amb activitat aciltransferasa. Diversos enzims pertanyen a aquesta família, segons la grandària de l’àcid gras emprat com a substrat: La carnitina acetiltransferasa (CrAT o CAT) utilitza acetil-CoA com a substrat. La carnitina octanoiltransferasa (CrOT o COT) exerceix el transport d’àcids grassos d’entre 8 i 10 carbonis entre els peroxisomes i els mitocondris Les carnitines palmitoiltransferases (CPT) 1 i 2 exerceixen el transport d’àcids grassos de cadena llarga (LC-CoA, de l’acrònim en anglès) d’entre 16 i 20 carbonis

[EN] Carbamoyl phosphate synthetase 1 (CPS1), a 1462-residue mitochondrial enzyme, catalyzes the entry of ammonia into the urea cycle, which converts ammonia, the neurotoxic waste product of protein catabolism, into barely toxic urea. The urea cycle inborn error and rare disease CPS1 deficiency (CPS1D) is inherited with mendelian autosomal recessive inheritance, being due to CPS1 gene mutations (>200 mutations reported), and causing life-threatening hyperammonemia. We have produced recombinantly human CPS1 (hCPS1) in a baculovirus/insect cell expression system, isolating the enzyme in active and highly purified form, in massive amounts. This has allowed enzyme crystallization for structural studies by X-ray diffraction (an off-shoot of the present studies). This hCPS1 production system allows site-directed mutagenesis and enzyme characterization as catalyst (activity, kinetics) and as protein (stability, aggregation state, domain composition). We have revealed previously unexplored traits of hCPS1 such as its domain composition, the ability of glycerol to replace the natural and essential CPS1 activator N-acetyl-L-glutamate (NAG), and the hCPS1 protection (chemical chaperoning) by NAG and by its pharmacological analog N-carbamyl-L-glutamate (NCG). We have exploited this system to explore the effects on the activity, kinetic parameters and stability/folding of the enzyme, and to test the disease-causing nature, of mutations identified in patients with CPS1 deficiency (CPS1D). These results, supplemented with those obtained with other non-clinical mutations, have provided novel information on the functions of three non-catalytic domains of CPS1. We have introduced three CPS1D-associated mutations and one trivial polymorphism in the glutaminase-like domain of CPS1, supporting a stabilizing and an activity-enhancing function of this non-catalytic domain. Two mutations introduced into the bicarbonate phosphorylation domain have shed light on bicarbonate binding and have directly confirmed the importance of this domain for NAG binding to the distant (in the sequence) C-terminal CPS1 domain. The introduction of 18 CPS1D-associated missense mutations mapping in a clinically highly eloquent central non-catalytic domain have proven the disease-causing nature of most of these mutations while showing that in most of the cases they trigger enzyme misfolding and/or destabilization. These results, by proving an important role of this domain in the structural integration of the multidomain CPS1 protein, have led us to call this domain the Integrating Domain. Finally, we have examined the effects of eight CPS1D-associated mutations, of one trivial polymorphism and of five non-clinical mutations, all of them mapping in the C-terminal domain of the enzyme where NAG binds, whereas we have re-analyzed prior results with another four clinical and five non-clinical mutations affecting this domain. We have largely confirmed the pathogenic nature of the clinical mutations, predominantly because of decreased activity, in many cases due to hampered NAG binding. A few mutations had substantial negative effects on CPS1 stability/folding. Our analysis reveals that NAG activation begins with a movement of the final part of the ß4-¿4 loop of the NAG site. Transmission of the activating signal to the phosphorylation domains involves helix ¿4 from this domain and is possibly transmitted by the mutually homologous loops 1313-1332 and 778-787 (figures are residue numbers) belonging, respectively, to the carbamate and bicarbonate phosphorylation domains. These two homologous loops are called from here on Signal Transmission Loops.
[ES] La carbamil fosfato sintetasa 1 (CPS1), una enzima mitocondrial, cataliza la entrada del amonio en el ciclo de la urea, que convierte esta neurotoxina derivada del catabolismo de las proteínas en urea, mucho menos tóxica. El déficit de CPS1 (CPS1D) es un error innato del ciclo de la urea, una enfermedad rara autosómica recesiva, que se debe a mutaciones en el gen CPS1 (>200 mutaciones descritas) y que cursa con hiperamonemia. Hemos producido CPS1 humana recombinante (hCPS1) en un sistema de expresión de células de insecto y baculovirus, y la hemos aislado en forma activa, muy pura y en cantidad elevada. Este sistema de producción de hCPS1 permite la realización de mutagénesis dirigida y la caracterización de la enzima como catalizador (actividad, cinética) y como proteína (estabilidad, estado de agregación y composición de dominios). Hemos revelado características de la hCPS1 antes no exploradas como es la composición de dominios, la capacidad que tiene el glicerol para reemplazar al activador natural y esencial de la CPS1, N-acetil-L-glutamato (NAG), y la protección de la hCPS1 por NAG y por su análogo farmacológico N-carbamil-L-glutamato (NCG) (chaperonas químicas). Hemos utilizado este sistema para explorar los efectos en actividad, parámetros cinéticos y estabilidad/plegamiento de la enzima, y para comprobar la naturaleza patogénica de mutaciones identificadas en pacientes con CPS1D. Estos resultados, junto con los obtenidos con otras mutaciones no clínicas, han aportado información novedosa sobre tres de los dominios no catalíticos de CPS1. Las observaciones realizadas tras introducir en el dominio de tipo glutaminasa de la enzima tres mutaciones asociadas a CPS1D y un polimorfismo trivial, apoyan la contribución de este dominio no catalítico a la estabilidad y a aumentar la actividad de la enzima. Dos mutaciones introducidas en el dominio de fosforilación de bicarbonato han arrojado luz sobre el modo de unión del bicarbonato (un sustrato). Los resultados de estas mutaciones también han confirmado la contribución de este dominio para la unión de NAG, cuyo sitio de unión se encuentra en el dominio C-terminal de CPS1, bastante alejado (en la secuencia) del dominio de fosforilación de bicarbonato. Además, hemos introducido 18 mutaciones de cambio de sentido asociadas a CPS1D, las cuales están localizadas en un dominio no catalítico, central y de elevada elocuencia clínica. Estos resultados han demostrado la naturaleza patogénica de estas mutaciones, ya que en la mayoría de los casos estas mutaciones producen un mal plegamiento o/y desestabilización de la enzima. Debido a que estos resultados han puesto de manifiesto el importante papel de este dominio en la integración estructural de la proteína multidominio CPS1, lo hemos llamado Dominio Integrador. Finalmente, hemos examinado los efectos de 8 mutaciones asociadas a CPS1D, de un polimorfismo trivial y de 5 mutaciones no clínicas, todas localizadas en el dominio C-terminal de la enzima, donde se une NAG. Además, hemos reanalizado resultados anteriores con otras 4 mutaciones clínicas y 5 no clínicas afectando a este dominio. Hemos confirmado el carácter patogénico de las mutaciones clínicas, las cuales predominantemente causan una disminución en la actividad enzimática, en muchos casos debida a que la unión de NAG se encuentra obstaculizada. Unas pocas mutaciones mostraron efectos negativos en la estabilidad/plegamiento de CPS1. Nuestros análisis revelan que la activación por el NAG empieza con un movimiento de la parte final del bucle ß4-¿4 del sitio de NAG. La transmisión de la señal activadora a los dominios de fosforilación implica a la hélice ¿4 de este dominio y posiblemente se transmite a través de los bucles homólogos 1313-1332 y 778-787 (numeración de residuos) pertenecientes, respectivamente, a los dominios de fosforilación de carbamato y bicarbonato. Por ello, hemos llamado a ambos bucles Bucles de
[CAT] La carbamil fosfat sintetasa 1 (CPS1), un enzim mitocondrial, catalitza l'entrada d'amoni en el cicle de la urea, que convertix l'amoni, producte neurotòxic del catabolisme de les proteïnes, en urea, una molècula molt poc tòxica. El dèficit de CPS1 (CPS1D) és un error innat del cicle de la urea, una malaltia rara autosòmica recessiva, que es deu a mutacions en el gen CPS1 (>200 mutacions descrites) i que cursa amb hiperamonièmia. Hem produït CPS1 humana recombinant (hCPS1) en un sistema d'expressió de cèl·lules d'insecte i baculovirus, i l'hem aïllada en forma activa, molt pura i en gran quantitat. Això ha permés la cristal·lització de l'enzim per a estudis estructurals amb difracció de raios-X (treball no inclòs en esta tesi Aquest sistema de producció de hCPS1 permet la realització de mutagènesi dirigida i la caracterització de l'enzim com a catalitzador (activitat, cinètica) i com a proteïna (estabilitat, estat d'agregació i composició de dominis). Hem revelat característiques de la hCPS1 no explorades abans com és la composició de dominis, la capacitat que té el glicerol per a reemplaçar l'activador natural i essencial de CPS1, N-acetil-L-glutamat (NAG), i la protecció de la hCPS1 per NAG i pel seu anàleg farmacològic N-carbamil-L-glutamat (NCG) (xaperones químiques) . Hem utilitzat aquest sistema per a explorar els efectes en l'activitat, els paràmetres cinètics i l'estabilitat/plegament de l'enzim, i per a comprovar la naturalesa patogènica de mutacions identificades en pacients amb CPS1D. Aquestos resultats, junt amb els obtinguts amb altres mutacions no clíniques, han aportat informació nova sobre tres dels dominis no catalítics de la CPS1. Les observacions, després d'introduir tres mutacions associades a CPS1D i un polimorfisme trivial en el domini tipus glutaminasa de CPS1, recolzen la contribució d'aquest domini no catalític a l'estabilitat i a l'optimització de l'activitat enzimàtica. Dues mutacions introduïdes en el domini de fosforilació de bicarbonat han esclarit el mode d'unió de bicarbonat. Els resultats d'aquestes mutacions també han confirmat la contribució d'aquest domini per a la unió de NAG, el lloc d'unió de la qual es troba en el domini C-terminal de CPS1, prou allunyat (en la seqüència) del domini de fosforilació de bicarbonat. A més, hem introduït 18 mutacions de canvi de sentit associades a CPS1D, les quals estan localitzades en un domini no catalític, central i d'elevada eloqüència clínica. Aquestos resultats han demostrat la naturalesa patogènica d'aquestes mutacions, ja que, en la majoria dels casos produïxen un mal plegament o/i desestabilització de l'enzim. Pel fet que aquestos resultats han posat de manifest l'important paper d'aquest domini en la integració estructural de la proteïna multidomini CPS1, l'hem anomenat Domini Integrador. Finalment, hem examinat els efectes de huit mutacions associades a CPS1D, un polimorfisme trivial i cinc mutacions no clíniques, totes elles localitzades en el domini C-terminal de l'enzim, on s'unix NAG. A més, hem reanalitzat resultats anteriors amb altres quatre mutacions clíniques i cinc no clíniques que afecten aquest domini. Hem confirmat el caràcter patogènic de les mutacions clíniques, les quals predominantment causen una disminució en l'activitat enzimàtica, en molts casos pel fet que la unió de NAG es troba obstaculitzada. Unes poques mutacions van mostrar efectes negatius substancials en l'estabilitat/plegament de CPS1. Les nostres anàlisis revelen que l'activació de NAG comença amb un moviment de la part final del bucle ß4-¿4 del lloc de NAG. La transmissió del senyal activadora als dominis de fosforilació involucra l'hèlix ¿4 d'aquest domini i es transmet, possiblement, a través dels bucles homòlegs 1313-1332 i 778-787 (numeració dels residus), pertanyents, respectivament, als dominis de fosforilació de carbamato i bicarbonat. Per això, hem anomenat a ambd
Díez Fernández, C. (2015). USING RECOMBINANT HUMAN CARBAMOYL PHOSPHATE SYNTHETASE 1 (CPS1) FOR STUDYING THIS ENZYME'S FUNCTION, REGULATION, PATHOLOGY AND STRUCTURE [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/52855
TESIS

Metabolites derived from dietary choline and L-carnitine, such as trimethylamine N-oxide and betaine, have recently been identified as novel risk factors for atherosclerosis in mice and humans. We sought to identify genetic factors associated with plasma betaine levels and determine their effect on risk of coronary artery disease (CAD). A two-stage genome-wide association study (GWAS) identified two significantly associated loci on chromosomes 2q34 and 5q14.1. The lead variant on 2q24 (rs715) localizes to carbamoyl-phosphate synthase 1 (CPS1), which encodes a mitochondrial enzyme that catalyses the first committed reaction and rate-limiting step in the urea cycle. Rs715 is also significantly associated with decreased levels of urea cycle metabolites and increased plasma glycine levels. Notably, rs715 yield a strikingly significant and protective association with decreased risk of CAD in only women. These results suggest that glycine metabolism and/or the urea cycle represent potentially novel sex-specific mechanisms for the development of atherosclerosis.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Os micro-organismos estão se tornando cada vez mais resistentes aos antimicrobianos e cepas de Candida albicans resistentes aos antifúngicos tem sido isoladas, assim, torna-se importante e necessário a realização de pesquisas que avaliem os efeitos de novos métodos terapêuticos, como a inativação fotodinâmica antimicrobiana (aPDI). Assim, o objetivo deste estudo foi verificar os efeitos da inativação fotodinâmica sobre biofilmes de Candida albicans, avaliando seus efeitos sobre a expressão dos genes TEC1 (fator de transcrição), HWP1 (proteína de parede celular das hifas), EFG1 (regulador transcricional relacionado com a morfogênese), BCR1 (regulador da formação de biofilme e da parede celular), CPH1 (regulador transcricional envolvido na morfogênese) e ALS3 (adesina) de C. albicans. Foram avaliadas 30 amostras isoladas de pacientes portadores de HIV e 30 amostras de pacientes com estomatite protética, quanto a produção de biofilme, peso seco e filamentação. Destas, foram selecionadas as amostras mais virulentas de cada grupo que apresentaram melhor capacidade de formação de biofilme e filamentação. Assim, foi utilizada uma amostra clínica de C. albicans isolada de paciente portador de HIV, uma amostra clínica de C. albicans isolada de paciente com estomatite protética e uma cepa padrão ATCC 18804. A quantificação da expressão dos genes foi relacionada à produção desses genes nas amostras clínicas e na cepa de referência utilizando-se ensaio de PCR em tempo real. Para a aPDI, foram utilizados os fotossensibilizadores azul de metileno a 300 μM e eritrosina a 400 μM sensibilizados com laser de Índio-Gálio-Alumínio-Fósforo de baixa potência (vermelho visível, 660 nm) e LED verde (532 ± 10 nm), respectivamente. Foram avaliados quatro grupos experimentais para a aPDI: a) F+L+: sensibilização com o corante e irradiação com luz; b) F+L-: somente tratamento com o fotossensibilizador; c) F-L+: somente irradiação com luz e d) F-L-: sem sensibilização com o corante e ausência de luz. Os resultados foram analisados por t-test, com um nível de significância de 5%. Após a análise fenotípica, as amostras Ca30 e 39S foram selecionadas para a realização da aPDI. Como esperado, apenas para o grupo F+L+, quando comparado com o grupo F-L-, todos os genes analisados foram sub expressos após a aPDI. O fold-decrease para os genes ALS3, HWP1, BCR1, TEC1, CPH1 e EFG1 foram 0,73; 0,39; 0,77; 0,71; 0,67 e 0,60; para laser, respectivamente, e 0,66; 0,61; 0,50; 0,43; 0,54 e 0,66; para LED, respectivamente. Pode-se concluir que a aPDI mostrou uma redução na expressão dos genes de C. albicans, sugerindo a diminuição de sua virulência.
Micro-organisms are becoming increasingly resistant to antimicrobial agents and Candida albicans resistant strains to antifungal has been isolated, so it is important and necessary to carry out studies that evaluates the effects of new therapeutic methods, such as antimicrobial photodynamic inactivation (aPDI). The objective of this study was verify the effects of aPDI on C. albicans biofilms, evaluating its effects on genes expression: TEC1 (transcription factor), HWP1 (cell wall protein hyphae), EFG1 (transcriptional regulator related to morphogenesis), BCR1 (regulator of biofilm formation and cell wall), CPH1 (transcriptional regulator involved in morphogenesis) and ALS3 (adhesin) of C. albicans. Were evaluated 30 samples isolated from patients with HIV and 30 samples from patients with denture stomatitis, as the production of biofilm, dry weight and filamentation. Of these, the most virulent strains of each group that presented better biofilm formation capacity and filamentation were selected. Therefore, were used a clinical sample of C. albicans isolated from HIV positive patient, a clinical sample of C. albicans isolated from patient with denture stomatitis and a standard strain ATCC 18804. The quantification of gene expression was related to the production of these genes in clinical samples and in the reference strain using PCR assay in real time. For aPDI, were used the photosensitizer methylene blue at 300 uM and erythrosine at 400 uM, sensitized with low power laser Indium-Gallium-AluminumPhosphorus (visible red, 660 nm) and green LED (532 ± 10 nm), respectively. Were evaluated four groups for aPDI: a) P+L+: sensitization with the photosensitizer and irradiation with light; b) P+L-: only treatment with the photosensitizer; c) P-L+: only irradiation with light and d) P-L-: without sensitization with the dye and absence of light. The results were analyzed by t-test, with a significance level of 5%. After the phenotypic analysis, the samples Ca30 and 39 S were selected for aPDI . As expected, only in the group P+L+ when compared with the group P-L-, all analyzed genes were downregulated after aPDI. The fold-decrease for the genes ALS3, HWP1, BCR1, TEC1, CPH1 and EFG1, were 0.73, 0.39, 0.77, 0.71, 0.67 and 0.60, for laser, respectively, and 0.66, 0.61, .050, 0.43, 0.54 and 0.66, for LED, respectively. It could be concluded that aPDI showed a reduction in the expression of C. albicans genes, suggesting its virulence decrease.
2013/22897-2

Orientador: Antonio Olavo Cardoso Jorge
Banca: Juliana Campos Junqueira
Banca: Graziella Nuernberg Back Brito
Banca: Martha Simões Ribeiro
Banca: Célia Regina Gonçalves e Silva
Resumo: Os micro-organismos estão se tornando cada vez mais resistentes aos antimicrobianos e cepas de Candida albicans resistentes aos antifúngicos tem sido isoladas, assim, torna-se importante e necessário a realização de pesquisas que avaliem os efeitos de novos métodos terapêuticos, como a inativação fotodinâmica antimicrobiana (aPDI). Assim, o objetivo deste estudo foi verificar os efeitos da inativação fotodinâmica sobre biofilmes de Candida albicans, avaliando seus efeitos sobre a expressão dos genes TEC1 (fator de transcrição), HWP1 (proteína de parede celular das hifas), EFG1 (regulador transcricional relacionado com a morfogênese), BCR1 (regulador da formação de biofilme e da parede celular), CPH1 (regulador transcricional envolvido na morfogênese) e ALS3 (adesina) de C. albicans. Foram avaliadas 30 amostras isoladas de pacientes portadores de HIV e 30 amostras de pacientes com estomatite protética, quanto a produção de biofilme, peso seco e filamentação. Destas, foram selecionadas as amostras mais virulentas de cada grupo que apresentaram melhor capacidade de formação de biofilme e filamentação. Assim, foi utilizada uma amostra clínica de C. albicans isolada de paciente portador de HIV, uma amostra clínica de C. albicans isolada de paciente com estomatite protética e uma cepa padrão ATCC 18804. A quantificação da expressão dos genes foi relacionada à produção desses genes nas amostras clínicas e na cepa de referência utilizando-se ensaio de PCR em tempo real. Para a aPDI, ... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Micro-organisms are becoming increasingly resistant to antimicrobial agents and Candida albicans resistant strains to antifungal has been isolated, so it is important and necessary to carry out studies that evaluates the effects of new therapeutic methods, such as antimicrobial photodynamic inactivation (aPDI). The objective of this study was verify the effects of aPDI on C. albicans biofilms, evaluating its effects on genes expression: TEC1 (transcription factor), HWP1 (cell wall protein hyphae), EFG1 (transcriptional regulator related to morphogenesis), BCR1 (regulator of biofilm formation and cell wall), CPH1 (transcriptional regulator involved in morphogenesis) and ALS3 (adhesin) of C. albicans. Were evaluated 30 samples isolated from patients with HIV and 30 samples from patients with denture stomatitis, as the production of biofilm, dry weight and filamentation. Of these, the most virulent strains of each group that presented better biofilm formation capacity and filamentation were selected. Therefore, were used a clinical sample of C. albicans isolated from HIV positive patient, a clinical sample of C. albicans isolated from patient with denture stomatitis and a standard strain ATCC 18804. The quantification of gene expression was related to the production of these genes in clinical samples and in the reference strain using PCR assay in real time. For aPDI, were used the photosensitizer methylene blue at 300 uM and erythrosine at 400 uM, sensitized with low power laser... (Complete abstract click electronic access below)
Doutor

Les entreprises manufacturières oeuvrant dans des domaines de haute technologie sont aujourd'hui confrontées à de multiples impératifs qui les incitent à collaborer avec des partenaires pour l'ensemble des leurs activités. Cette étude analyse l'impact de la planification de la collaboration sur la performance d'une chaîne d'approvisionnement. L'étude a été menée en deux phases dans la chaîne d'approvisionnement d'un équipementier du secteur des télécommunications. La première phase, l'étude de terrain, a comme objectifs de bien cerner les méthodes de collaboration et d'identifier les intérêts des différents membres de la chaîne d'approvisionnement. La deuxième phase, le questionnaire électronique, a comme objectifs d'analyser l'impact de la planification de la collaboration sur l'innovation au sein des entreprises de la chaîne d'approvisionnement, et d'évaluer l'effet médiateur des innovations sur la relation entre la planification de la collaboration et la performance de la chaîne.

En el presente estudio se intentó incrementar el proceso de reducción de disonancia cognitiva a través de la estimulación de la corteza pre-frontal (CPF) izquierda por contracciones con la mano contraria (la derecha). Se esperó que mayor actividad en un hemisferio de la CPF se reflejara en un aumento de la temperatura de la membrana timpánica (TMT), del hemisferio estimulado. Sin embargo, no se observó cambio alguno en la TMT opuesta a la mano que realizó las contracciones. Por el contrario, se observó una disminución de la TMT ipsilateral a la mano estimulada. Asimismo, no hubo diferencias en el proceso de reducción de disonancia entre aquellos que realizaron contracciones con la mano derecha y aquellos que realizaron contracciones con la mano izquierda.
The present study attempted to increase cognitive dissonance reduction stimulating the left prefrontal cortex (PFC) by unilateral right hand clenching. More activity in the left PFC was expected to increase the ipsilateral tympanic membrane temperature (TMT). However no change was observed in the TMT opposite to the clenched hand. Contrary to the expected, a decrease of the TMT ipsilateral to the hand clenched was observed. No differences in dissonance reduction were observed between experimental groups.
Tesis

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CAPS - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
RESUMO Por meio da presente dissertação, apresento os encadeamentos do processo de encenação que aconteceram no Conjunto Penal Feminino de Salvador (CPF), cartografados na travessia entre o teatro e o presídio, com o intuito de investigar as interações entre o espaço físico do sistema prisional, suas disciplinas e circunstâncias normativas, com vistas à ideação da existência de um lugar onde as fronteiras são permeáveis, constituindo uma zona de transgressão, não restrita ao espaço físico, mas resultante da articulação do espaço das diversidades, um espaço dinâmico, espaço “de e para” criação, onde somos confrontados com os nossos próprios limites. Assim, discorro sobre quem penso que sou, porque escrevi esta dissertação e como materializei essa produção, em parte dialética e em parte subjetiva, na qual me coloco na posição de sujeito errante de uma trajetória rizomática. O propósito foi discutir “no e sobre” o presídio, um lugar de encontros e desencontros, lugar de relações com um outro que poderia ser nós mesmos. A partir das formulações de Lemgruber, Goffman, Foucault, Carvalho Filho, Maia e colaboradores, acerca do poder, controle e vigilância das instituições totais, das consequências e privações do encarceramento, destaco aspectos percebidos que poderiam despir noções e preconceitos a respeito de quem está presa ou preso e com isso alterar configurações do pensamento de quem está fora dos muros dos cárceres. Nesse trajeto, os princípios e técnicas do TO forneceram considerável suporte para o desenvolvimento das oficinas e, junto com os indutores do jogo proposto por Ryngaert, propiciaram a elaboração de proposições que me levaram a compreender o descobrir/saber/fazer teatro em situações adversas. A dissertação divide-se em três pistas que contêm chaves nas quais discuto questões específicas do presídio, do método e da oficina de teatro realizada no CPF. Assim, reflito sobre o presídio como instituição total e molar detentora do tempo de pessoas e sobre as consequências do encarceramento; sobre o processo como método da pesquisa, mapeando tudo que pensava em conjunção com as leituras, experimentos, músicas e as vozes que ecoavam, pois precisava visualizar os territórios por onde me deslocava e, sendo assim, elaborei mapas visuais para entender as possíveis conexões de força entre eles e os dispositivos de pergunta/resposta/pergunta a partir dos quais fagulhas do pensamento de Deleuze e Guattari sustentaram o trajeto e as possibilidades de cruzamento que dele surgia. Descrevo as singularidades da criação no Conjunto Penal Feminino – Salvador, que foram divididos no período do Projeto Dialogando com a Liberdade em 2014 e durante o reencontro com às mulheres presas, que aconteceu em 2017/2018, período desta investigação. Assim, aproximei e ao mesmo tempo delimitei as fronteiras entre o teatro e o presídio (liberdade e encarceramento), para encadeá-las na escrita desta dissertação. Deste modo, nas considerações nada finais concebo o teatro como ato de resistência que projetou a voz das mulheres presas.
RÉSUMÉ Grâce à cette thèse, je présente les fils du processus de mise en scène qui a eu lieu dans le création la Institution pénale feminine de Salvador (CPF), mis en correspondance à la frontière entre le théâtre et la prison, afin d'étudier les interactions entre l'espace physique du système pénitentiaire, leurs disciplines et les circonstances réglementaires, en vue de l'idéation de l'existence d'un lieu où les limites sont perméables, constituant une zone de transgression, ne se limite pas à l'espace physique, mais en raison de l'espace commun de la diversité, un espace dynamique, l'espace pour la création, où nous sommes confrontés à nos propres limites. Donc, je parle au sujet de qui je pense que je suis, le pour quoi j'ai écrit cette thèse et comment cette production a etait matérialisée, dans la dialectique de la pièce et une partie subjective, où je me place dans la position du sujet errant d'une trajectoire de rhizome. Le but était de discuter “à l'intérieur et a propos” de la prison, un lieu d'accords et désaccords, un lieu de rencontre avec ceux qui pourraient être nous-mêmes. A partir de conception Lemgruber, Goffman, Foucault, Carvalho Filho, Maia et collaborateus, sur le pouvoir, le contrôle et la surveillance des institutions totales, les conséquences et l'emprisonnement de privation, et mettre en évidence les aspects qui pourraient dépouiller les notions et les préjugés au sujet de qui est arrêtés ou emprisonnés et altérant ainsi la pensée de ceux qui sont en dehors des murs de la prison. Dans cette voie, les principes et les techniques du théâtre de l'opprimé on fourni un soutien considérable pour le développement des ateliers et en collaboration avec les inducteurs du jeu proposé par Ryngaert, a conduit à l'élaboration de propositions qui m'a amené à comprendre la découverte / savoir / faire du théâtre dans des situations difficiles. La thèse est divisée en trois pistes qui contiennent des clés sur des questions spécifiques qui traitent de la prison, la méthode et l'atelier de théâtre a eu lieu à CPF. Ainsi, je réfléchis à la prison en tant qu’institution totale et molaire détenant le temps des personnes et aux conséquences de l’incarcération; au sujet du processus en tant que méthode de recherche, la cartographie toute pensée en conjonction avec les lectures, les expériences, la musique et les voix qui résonnaient, pour visualiser les territoires où me déplacés, par conséquent jai’s élaboré des cartes visuelles pour comprendre les possibles connexions de force entre eux et les techniques de question / réponse / question qui suscite la pensée de Deleuze et Guattari ont soutenu le chemin et les possibilités de le traverser. Je décris les singularités de la création dans le cadre pénal féminin - Salvador, qui ont été divisées pendant la période du projet Dialogando com a Liberdade en 2014 et lors de la rencontre avec les femmes détenues en 2017/2018, période de l'enquête. Ainsi, je me suis approché et en même temps délimité les frontières entre le théâtre et la prison (liberté et emprisonnement), pour les relier à la rédaction de cette thèse. Ainsi, dans les dernières considérations, je conçois le théâtre comme un acte de résistance qui projette la voix des femmes détenues.

碩士
國立臺灣大學
生化科學研究所
107
Mitochondria are power generators in eukaryotic cells, but the oxidative reaction makes mitochondrial DNA (mtDNA) more susceptible to damages and leads to mutations. More than 50 diseases are caused by mtDNA mutations, and they are difficult to study and nearly impossible to cure. Programmable nucleases such as ZFN and TALEN have been used to target mtDNA (mostly for depletion of mutant mtDNA), but the design and construction are labor-intensive and time-consuming. By contrast, RNA-guided CRISPR gene technology is simpler. In this thesis, I aimed to engineer CRISPR-Cpf1 for human mtDNA editing. CRISPR-Cpf1 system has several advantages over CRISPR-Cas9. Cpf1 has shorter crRNA that may allow better entry into the mitochondria. In addition, Cpf1 has RNA nuclease activity that can process an array of crRNAs into individual crRNAs of the same or multiplex target sequences. Finally, Cpf1 cleavage generates stagger cuts to potentially allow DNA insertion via microhomology-mediated end joining (MMEJ), which is reported to be the only active double-strand break (DSB) repair pathway in human mitochondria. To reach and edit mtDNA in the mitochondrial matrix, it is necessary to engineer mitochondria-targeting Cpf1 and crRNA (termed mito-Cpf1 and mito-crRNA, respectively). To achieve this, I have screened several protein and RNA mitochondrial targeting sequences, and validated the localization of mito-Cpf1 and mito-crRNA by mitochondrial extraction, Western blotting, immunofluorescent microscopy and RT-qPCR. My next step was to reconstitute active Cpf1 complexes in mitochondria to target and cleavage specific sites on mtDNA. My goal was to detect evidence of mtDNA cleavage and subsequent reduction in mitochondrial oxidative respiration. A robust CRISPR-based mtDNA editing system would open many possibilities to study mtDNA maintenance and to pave the path for therapeutic editing of mtDNA mutations.

碩士
國立臺北科技大學
機電整合研究所
95
In the competitive environment of global supply chain at this present, in order to satisfy more and more demands and expectations from customers, enterprises face many pressures such as cost reduction, short lead-time, quality promotion, and global operation management…etc. But enterprises can’t resolve this pressure by their own; instead, it shall be achieved by the collaboration and integration of overall business entities of supply chain based on the same targets. The VICS committee has published the standards for Collaborative Planning, Forecasting and Replenishment (CPFR), and CPFR has the revolutionary influence on supply chain. However, according to the relevant research and investigation, the applications of CPFR in Taiwan are not prevalent yet currently. Most of enterprises still have the issue of forecast and replenishment with their business entities, and can not reduce cost and shorten lead time effectively. The major cause is enterprises haven’t implemented a CPFR system to link and coordinate overall business entities of supply chain. According to this, this research are based on the enterprise’s demands on CPFR system and literatures review, and use seven-stages development methodology which are including define objectives, process modeling, data modeling, apply business rules, UI interface design, coding and testing. Then focus on the key stages of define objectives and process modeling, and provide a collaborative process modeling that is suitable for overall business entities of supply chain (including four organizations: global operation, subsidiary, factory and buyer). Hopefully, the process modeling will help who are planning to implement a CPFR system. Besides, in order to promote the service capability of orders with multiple deliveries, the process modeling in this research integrate the technology of real-time CTP to shorten lead time and reduce cost based on the well operation of capacity and inventory.

碩士
國立政治大學
資訊管理研究所
96
CPFR (Collaborative Planning, Forecasting, and Replenishment) is one of the applications of collaborative business. The stressed concept is the cooperation process of sellers and buyers on the supply chain in order to increase the handling efficiency. In the future, the industries would compete on the whole supply chains behind products—only the industry that is capable of making accurate predictions according to the constantly changing market and reacts immediately has the chance of winning. Being able to control the inventory and supply effectively would be one of the key factors leading to an industry’s success. The replenishment model of CPFR is to fill out the order according to the sales prediction, order prediction, inventory strategy, and supply information. The precision of the replenishment model could affect both suppliers and customers. The former can distribute products properly and meet the different demands from the upcoming orders so as to reduce inventory; the latter are able to revise the inventory strategy and amount of order according to the order prediction. A few research papers aimed at the replenishment model, though, most still focus on the management issues like the process framework of CPFR and the implementation benefit. Hence, establishing both an information system that coordinates customer demand with suppliers and a collaborative replenishment model that increases the accuracy of predictions is fairly important. The phase of replenishment, as the subject of this study, will approach on parameters the collaborative replenishment model needs to input and combine evolution strategies with tabu search to establish a replenishment model under the process of CPFR.

碩士
東吳大學
企業管理學系
97
In the fast-changing industry enviroment, enterprises have paid a lot of money to invest new technology and new market to gain their own competition advantage. But the new technology progressing and material cost rasing make enterprises faced more difficult business competition. Therefore, the various strategic alliance and cooperation are established. This research is to make the in-depth case analysis for three MFP makers how to cooperate with CIS suppliers and enhance their operational agility by CPFR (Collaborative Planning, Forecasting,and Replenishment ) with information sharing。 We can investigate and survey their individual cooperating result. Usually, the data of order forecasting exists uncertainly and vagueness which forced CIS suppliers to adjust their productin plan to catch the tight delivery, and the replenishment system of JIT and VMI caused the high inventory and low material cycling rate. Therefore, Rolling Forecasting has become a well-known and criticial solution. The research reveals some valuable sugestions are the best model of the order forecasting is to implement“VMI + Rolling Forecasting” system. It is helpful to decrease the high inventory & transportation cost to improve the supply chain performace. This model also can be a reference case for other industries.

碩士
開南大學
物流與航運管理學系
98
In recent years, supply chain management has become an essential business strategy. When the enterprise, supplier and customer construct a supply chain of production and marketing cooperating relationship, to enhance sharing information would make the operating integration more smoothly. After financial tsunami, hypermarkets are seeking approaches to increase sales. Besides, under the premise that the effect of the attitude of consumer spending cautiously makes businesses have to use various strategies to attract the consumers to purchase products. For this reason, retailers and wholesalers devoted to develop private brand (also named store brand) which can keep profits and attract consumers by favorable price. This study adopts system dynamics to simulate the operation of private brand products under CPFR (Collaborative Planning, Forecasting, and Replenishment) model between the hypermarket and the supplier. The aim is to discuss whether the CPFR model can raise the operating performance or not, and also try to understand how the suppliers and hypermarkets to create win-win situation in a cooperative method. Applying CPFR model can not only make hypermarket maintain lower stocks and reduce the potential loss of stock, but also minimize all the uncertainty of supply and demand; moreover, bringing the benefits of lower costs and increase sales. Three types of private brand products are selected to compare with each other in this study for the sake of learning the cost and customer satisfaction under the CPFR model; furthermore, providing advices of the implementation of priority order.

碩士
國立高雄應用科技大學
工業工程與管理系碩士班
93
VICS association proposed the Collaborative Planning, Forecasting and Replenishment (CPFR ) model in 1998, it is one of the latest and important ways in the supply chain collaboration management field. Through the nine steps of the CPFR that will lead the forecasting in the collaborative supply chain to be more promissory, and let the replenishment be more efficient. In order to let CPFR be carried out smoothly, this research set the development cycle of the Key performance index, develops the information sharing mechanism of the across organization, and constructs collaborative management by exception mechanism model. Through the three core operation mechanisms constructed in this research that can help enterprises when inject into CPFR to be depend on operation procedure followed, in order to let enterprises succeed while carrying out the CPFR. A shaft bearing company is considered as case study, through the case company to verify the three core operation mechanisms in CPFR which constructed in this research are able to be carried out. Besides, this research can provide the case company to understand the implementation structure and procedure of the whole CPFR even more. At the same time , according to the detail operation procedure of three core operation mechanisms which constructed in this research could to remedy and strengthen stipulating in KPI of case company, could help the case company to choice the information sharing partners in the supply chain and remedy the deficiency of the handling exception mechanism. In general, the three core operation mechanisms lead the case company to execute the CPFR going smoothly, and make it obtain success.

碩士
國立臺北科技大學
製造科技研究所
97
Because the global supply chain competition, enterprises are facing pressure on the operation must be by members of the supply chain collaboration to address the pressure. CPFR has been made, and foreign enterprises also confirmed the validity of the solution. To make it easier for domestic companies to import CPFR, the research enterprise into CPFR can be reference to the business model. In order to avoid the application of business models fail because supply chain members do not know the scope of work and working order, clear definition of each role is responsible for the content of the work and the work order, from the definition of the VICS standard CPFR processes carried out 22 kinds of group work. In addition, to enable the successful completion of work, the research and analysis of electronic systems CPFR functional structure of modules, a total of five modules. However, when the use of members of the supply chain changes, the business model of the processes will change. Therefore, this study analyzes the operation of domestic enterprises, according to the operation of domestic enterprises and demand characteristics, identifies the three members of the supply chain combinations, respectively, for manufacturers - traders - distributors, suppliers - brand business - distributors, manufacturers - their own brand retailers. To prove the proposed business model is feasible, to the second supply chain portfolio analysis as a case, CPFR applications into the company''s business model, operation of the system the screen showed. And analysis of the supply chain model of the other two cases the difference between applications in order to facilitate the supply chain model different enterprise applications of the model.