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Understanding the Management Control System Used by Hybrid Companies : A Case Study of IKEA and MIO.Chaussée, Laury, Foray, Marine January 2018 (has links)
Due to the globalization, companies have to review their strategy permanently. In the same way, they need to choose the right management control system which will be in line with their strategy. Recent research has been made and it was highlighted that new strategies have been created to face up to the globalization and stay competitive. One of the new strategies is called “hybrid strategy”. It consists of combining the low-cost competitive advantage with the differentiation competitive advantage. By doing that, a company differentiates itself from its competitors with a special feature and, by the same time, has the lowest cost in its market. This new strategy came after the three generic strategies from Porter, which explains that a company needs to choose a competitive advantage if it wants to be profitable. The competitive advantages are either low-cost or differentiation. Most of the research made about the hybrid strategy was about the efficiency of this strategy. A very few research was made about the management of companies using the hybrid strategy. This is why in this paper, we would like to understand what kind of management control companies using this strategy have, more particularly do they use more a tight or a loose control. We also are looking at what kind of objectives companies using hybrid strategies have. If it is more a short-term or a long- term perspective and if it is more focused on monetary goals or non-monetary goals. For that, we compared the management control implemented in a hybrid company with the management controlled implemented in a low-cost company in Sweden. Four interviews were made at IKEA and MIO, which are two Swedish furniture companies in Sweden. To be able to draw a conclusion, we choose one hybrid company and one pure company to compare them. IKEA is the company using the hybrid strategy. These interviews gave us a lot of information to better understand the type of management control both companies have implemented. Then, the data were analyzed and discussed in order to highlight a potential conclusion. The gathered data suggest that hybrid companies and companies using a low-cost strategy have approximately the same management control system which is more tight control. About the objectives, both companies have the same. Indeed, IKEA and MIO have monetary goals and are more short-term goals oriented, at least for the lower level management. However, there are some differences that we can noticed. The first thing is that MIO uses more the informal control than IKEA. The second thing is that IKEA is more concerned by its employees. To conclude, even if IKEA and MIO are using two different strategies, they are very similar when looking to their management control.
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NUEVAS ESTRATEGIAS PARA INCREMENTAR LA CALIDAD NUTRICIONAL DE PRODUCTOS DE PANADERÍA. EFECTO SOBRE EL CONTENIDO DE FITATOS Y LA BIODISPONIBILIDAD DE HIERRO EN CACO-2Sanz Penella, Juan Mario 30 March 2012 (has links)
Los productos de cereales con grano entero, en particular el pan, son fuentes de fibra dietética, vitaminas, minerales y compuestos fitoquímicos. Sin embargo, la biodisponibilidad de los minerales se afecta principalmente debido a la presencia de fitatos que reducen su absorción. El fitato se encuentra principalmente en los alimentos no procesados, pero puede ser degradado durante la germinación de las semillas o el procesado de alimentos. La hidrólisis del fitato a fosfatos de mio-inositol de menor grado de fosforilación es una manera de reducir su efecto negativo sobre la absorción de minerales. Muchas investigaciones han intentado reducir la cantidad de fitatos en los alimentos mediante diferentes procesos o adición de fitasas exógenas.
El objetivo principal de esta investigación fue incrementar el valor nutricional de los productos derivados de cereales a través de nuevas estrategias para la elaboración de productos de panadería de alta calidad, proporcionando más fibra dietética y una mayor biodisponibilidad de minerales. La inclusión de salvado de trigo en diferentes niveles y tamaño de partícula, con ?-amilasa y fitasa; el uso de bifidobacterias productoras de fitasa como nuevos iniciadores panarios; y la utilización de harina integral de amaranto como ingrediente nutritivo en panificación, fueron las estrategias propuestas para alcanzar el objetivo principal. Se evaluó la calidad nutricional, tecnológica y sensorial de los productos desarrollados. Asimismo, se estudió el efecto de la formulación en la relación molar fitato/mineral, la dializabilidad del hierro y la biosíntesis de ferritina en células Caco-2 como una medida de la absorción de este mineral. El salvado de trigo, en combinación con enzimas amilolíticas y fitasa, disminuyó el efecto negativo en la reología del producto y mejoró la hidrólisis de los fitatos. El uso de bifidobacterias productoras de fitasa (GRAS/PQS), tanto en proceso directo como indirecto, produjo panes con características tecnológicas y sensoriales similares a
los controles, pero con una cantidad menor de fitatos. Se consiguió una proporción
de compromiso de harina de amaranto en la formulación de pan para preservar la
calidad del producto e incrementar su valor nutricional. / Sanz Penella, JM. (2012). NUEVAS ESTRATEGIAS PARA INCREMENTAR LA CALIDAD NUTRICIONAL DE PRODUCTOS DE PANADERÍA. EFECTO SOBRE EL CONTENIDO DE FITATOS Y LA BIODISPONIBILIDAD DE HIERRO EN CACO-2 [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/15151 / Palancia
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El nucleo familiar cidiano : su funcion dentro del medievo espanol en el "Cantar de mio Cid"Garcia, Martha 01 April 2000 (has links)
No description available.
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Modulation of the Progenitor Cell and Homeostatic Capacities of Müller Glia Cells in Retina : Focus on α2-Adrenergic and Endothelin Receptor Signaling SystemsHarun-Or-Rashid, Mohammad January 2016 (has links)
Müller cells are major glial cells in the retina and have a broad range of functions that are vital for the retinal neurons. During retinal injury gliotic response either leads to Müller cell dedifferentiation and formation of a retinal progenitor or to maintenance of mature Müller cell functions. The overall aim of this thesis was to investigate the intra- and extracellular signaling of Müller cells, to understand how Müller cells communicate during an injury and how their properties can be regulated after injury. Focus has been on the α2-adrenergic receptor (α2-ADR) and endothelin receptor (EDNR)-induced modulation of Müller cell-properties after injury. The results show that α2-ADR stimulation by brimonidine (BMD) triggers Src-kinase mediated ligand-dependent and ligand-independent transactivation of epidermal growth factor receptor (EGFR) in both chicken and human Müller cells. The effects of this transactivation in injured retina attenuate injury-induced activation and dedifferentiation of Müller cells by attenuating injury-induced ERK signaling. The attenuation was concomitant with a synergistic up-regulation of negative ERK- and RTK-feedback regulators during injury. The data suggest that adrenergic stress-signals modulate glial responses during retinal injury and that α2-ADR pharmacology can be used to modulate glial injury-response. We studied the effects of this attenuation of Müller cell dedifferentiation on injured retina from the perspective of neuroprotection. We analyzed retinal ganglion cell (RGC) survival after α2-ADR stimulation of excitotoxically injured chicken retina and our results show that α2-ADR stimulation protects RGCs against the excitotoxic injury. We propose that α2-ADR-induced protection of RGCs in injured retina is due to enhancing the attenuation of the glial injury response and to sustaining mature glial functions. Moreover, we studied endothelin-induced intracellular signaling in Müller cells and our results show that stimulation of EDNRB transactivates EGFR in Müller cells in a similar way as seen after α2-ADR stimulation. These results outline a mechanism of how injury-induced endothelins may modulate the gliotic responses of Müller cells. The results obtained in this thesis are pivotal and provide new insights into glial functions, thereby uncovering possibilities to target Müller cells by designing neuroprotective treatments of retinal degenerative diseases or acute retinal injury.
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O foot-ball de todos: uma hist?ria social do futebol em Porto Alegre, 1903-1918Soares, Ricardo Santos 26 March 2014 (has links)
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Previous issue date: 2014-03-26 / Before becoming the most practiced sport of the present day, football experienced a broad and complex process of spatial diffusion, coming from Victorian England towards the most diverse regions of the planet. Characterized as a sport of the elite, it arrives in Brazil in this same condition, however it experiences in Rio Grande do Sul a unique "appropriation". Pointing out the differences in origin in comparison with the rest of the country, this work focuses on the particularity that was historically translated as a "construction of national identity," which in the south of Brazil also emerges by the hands of the elite, who was composed of teutos immigrants. At the same time the sport is rapidly absorbed by a portion of the city s black community. The outcome of this research unveiled the importance of the black ethnicity, which combined with the elite and the large number of immigrants, introduces a type of football to Porto Alegre with an important role in the process of ideological identification. This process turns football players into representative symbols of social groups. It has also been concluded as a historical product, that there has been a new existent relationship of identification between the spectators and their clubs, giving birth to supporters. In this sphere, Gremio and Internacional are born, an event which will later be appropriated by the capitalist system, taking this "passion" and turning it into a commodity. For its feasibility the research points to the necessity of a broad debate about the suitability of football in the business world, while suggesting in this context the role of historians. This role should not be to legitimize fabricated traditions, but rather to provoke a critical analysis of this highly popular sport, avoiding the subterfuge of shallow narratives and allowing the chronicle to exceed its limits. / Antes de tornar-se a modalidade esportiva mais praticada na atualidade, o futebol vivenciou um amplo e complexo processo de difus?o espacial, partindo da Inglaterra vitoriana em dire??o ?s mais diversas regi?es do planeta. Caracterizado como atividade da elite, chega ao Brasil nessa mesma condi??o, todavia experimenta no Rio Grande do Sul uma apropria??o singular. Pontuando as diferen?as de origem com o resto do pa?s, o trabalho privilegia a especificidade que se traduziu historicamente como uma constru??o de identidade nacional, e que, no sul, chega tamb?m pelas m?os da elite, que, por sua vez, era composta por imigrantes teutos, ao mesmo tempo em que a modalidade ? rapidamente absorvida por uma parcela da comunidade negra da cidade. Os resultados que emergiram, desvelaram a import?ncia da etnia negra, que agregada ? elite e ao grande contingente de imigrantes, inaugura um futebol em Porto Alegre com important?ssimo papel no processo de identifica??o ideol?gica, que transforma jogadores em representantes simb?licos de grupos sociais. Tamb?m concluiu-se existir, como produto hist?rico, uma rela??o nascente de identifica??o dos espectadores com os clubes nascendo portanto os torcedores. Neste universo, emergem Gr?mio e Internacional, fato que ser? apropriado pelo sistema capitalista, tomando essa paix?o e mercantilizando suas express?es. Para sua viabilidade, a pesquisa aponta para a necessidade de um amplo debate sobre uma adequa??o do futebol ao mundo dos neg?cios, ao mesmo tempo em que sugere, nesse contexto, o papel de vigil?ncia do historiador para n?o legitimar mais tradi??es inventadas. Antes disso, deve provocar uma reflex?o cr?tica desse esporte t?o popular, escapando do subterf?gio, da cr?nica, possibilitando a ultrapassagem dos limites da narra??o.
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Ensaios em economia da migra??o : uma an?lise de padr?es migrat?rios no BrasilRibeiro, Ana Carolina Borges Marques 11 July 2017 (has links)
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Previous issue date: 2017-07-11 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / This thesis presents three independent essays in Economics of Migration. At first, the
aim is to measure the impact of an international migration experience on wages in
the Brazilian labor market in 2010. The estimation becomes more complex due to the
various potential selection biases involved. Compare returned migrants to nonmigrant
individuals without regard the selection processes involved can generate
biased estimates of the impact of the migration experience on wages. The main
contribution of this study is to estimate the wage premium for returnees to Brazil
considering a triple selection process. I estimated a model of simultaneous equations
considering the occupation decision, the decision to participate in the labor market,
and the decision to return to the country since the individual had previously
emigrated. Even after controlling for potential biases, the results indicate that there is
a positive and statistically significant wage premium for the migration experience on
wages, indicating that return migration generates benefits to the country by bringing
individuals with characteristics valued in the labor market of the country, and,
therefore, this population can be a channel of increase of human capital and
productivity in Brazil. The second essay analyzes how the human capital and the
income of the spouses can be associated to the family migration in Brazil. We used
microdata from the 2010 Census to estimate a logit model, where three groups of
families are investigated: non-migrants, in-state migrants, and interstate migrants.
The causal effect of the variables was not analyzed, but only its association with the
probability of family migration. The contribution of the study is empirical; the results
show that the fact that the spouses have higher education favors the propensity to
migrate, but the employment relationship of the tied spouse (the one whose wage
variation does not determine the migration), usually the woman, has a negative
association with the probability of migration. When analyzing the income variables,
the evidence shows that both a greater income dispersion between the spouses and
a higher sum of the spouses' incomes have a positive association with the probability
of migration. When comparing only migrant families, the results indicate that the
probability of interstate migration is not associated with a higher educational level of
the spouses. Finally, the third essay analyzes the role of migration networks in
migratory flows in Brazil for different levels of schooling. A network of migrants can
be seen as a social network that attracts new migrants because of the network's informational and financial support that can reduce migration costs and facilitate new
migration flows. As migration costs are relatively higher for low-skilled individuals,
one hypothesis is that migration networks reduce the qualification levels of new
migration flows, making negative self-selection more likely. The objective of this study
is to analyze the role of migration networks in migratory flows within Brazil. The
results show a positive and significant impact of migrant networks on migratory flows
of different levels of schooling. In addition, the selection of migrants is also influenced
by migrant networks, which particularly attract new low-skilled migrants. The results
are robust to various econometric specifications, including the treatment of selection
bias and endogeneity. / Esta tese apresenta tr?s ensaios independentes em Economia da Migra??o. O
objetivo do primeiro ensaio ? mensurar o impacto da experi?ncia de migra??o
internacional sobre sal?rios no mercado de trabalho brasileiro em 2010. A estima??o
se torna mais complexa devido aos v?rios potenciais vieses de sele??o envolvidos.
Comparar emigrantes retornados a indiv?duos n?o migrantes sem levar em conta os
processos de sele??o envolvidos pode gerar estimativas distorcidas do impacto da
experi?ncia de migra??o sobre sal?rios. A principal contribui??o deste estudo ?
estimar o pr?mio salarial para retornados ao Brasil considerando um triplo processo
de sele??o. Foi estimado um modelo de equa??es simult?neas considerando a
decis?o de ocupa??o, a decis?o de participa??o no mercado de trabalho, e a
decis?o de retorno ao pa?s dado que, o indiv?duo emigrou anteriormente. Mesmo
ap?s controlar os potenciais vieses, os resultados mostram que h? um pr?mio
salarial positivo e estatisticamente significante da experi?ncia de migra??o sobre
sal?rios, indicando que a migra??o de retorno gera benef?cios ao pa?s ao trazer
indiv?duos com caracter?sticas valorizadas no mercado de trabalho local, e, portanto,
esta popula??o pode ser um canal de aumento de capital humano e produtividade
no Brasil. O segundo ensaio analisa como a renda e o capital humano dos c?njuges
podem estar associados ? migra??o familiar no Brasil. Foram utilizados microdados
do Censo de 2010 para estimar um modelo logit, onde se investiga a migra??o de
tr?s grupos de fam?lias: as n?o migrantes, as migrantes intra-estaduais e as
migrantes interestaduais. N?o foi analisado o efeito causal das vari?veis, mas
apenas sua associa??o com a probabilidade de migra??o familiar. A contribui??o do
estudo ? emp?rica; os resultados mostram que o fato dos c?njuges possu?rem ensino
superior favorece a propens?o a migrar, mas o v?nculo laboral do c?njuge vinculado
(aquele cuja varia??o salarial n?o determina a migra??o), em geral a mulher, tem
uma associa??o negativa com a probabilidade de migra??o. Quando se analisa as
vari?veis de renda, as evid?ncias mostram que tanto uma maior dispers?o de renda
entre os c?njuges, quanto uma maior soma das rendas dos c?njuges, tem uma
associa??o positiva com a probabilidade de migra??o. Quando s?o comparadas
apenas as fam?lias migrantes, os resultados indicam que a probabilidade de
migra??o interestadual n?o est? associada a um maior n?vel educacional dos
c?njuges. Por fim, o terceiro ensaio apresenta uma an?lise do papel das redes de migra??o nos fluxos migrat?rios no Brasil para diferentes n?veis de escolaridade.
Uma rede de migrantes pode ser vista como uma rede social que atrai novos
migrantes. O suporte informacional e financeiro desempenhado pela rede pode
diminuir os custos de migra??o e facilitar novos fluxos migrat?rios. Como os custos
de migra??o s?o relativamente maiores para os indiv?duos de baixa qualifica??o,
uma hip?tese ? que as redes de migra??o reduzem os n?veis de qualifica??o dos
novos fluxos migrat?rios, tornando uma autossele??o negativa mais prov?vel. O
objetivo do estudo ? analisar o papel das redes de migra??o nos fluxos e na sele??o
de migrantes dentro do Brasil. Os resultados mostram um impacto positivo e
significante das redes de migrantes nos fluxos migrat?rios de diferentes n?veis de
escolaridade. Al?m disso, a sele??o de migrantes tamb?m ? influenciada pelas
redes de migrantes, que particularmente atraem novos migrantes de pouca
qualifica??o. Os resultados s?o robustos a diversas especifica??es econom?tricas,
incluindo o tratamento de vi?s de sele??o e endogeneidade.
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Dois ensaios em economia internacional e mercado de trabalhoGranitoff, Igor 27 March 2018 (has links)
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Previous issue date: 2018-03-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The present dissertation contains two essays about implications of international trade on labor market. In the first essay the aim is to find correlations between medium wage of firms and the fact of these firms export or not. It was intend to answer if medium wage of exporting firms differentiate from the not exporter ones, if the level of education of the human capital of the firms play a hole on the premiums for export and if the level of development of the destiny of the exportations imply in any way the magnitude of these premiums. Results showed that there is a wage premium for export in firms that send its production to high developed countries. To the other categories of development, there is a correlation between major levels of education of the human capital of the firms and exporting to high competition markets analyzes the impact of international trade on the differential of wages between genders in Brazil. The second essay objectives to seek evidence of contribution of the fact that a firm is in the international market (through the export of its products) to soften the existing gender wage-gap in Brazil. Purposed to answer if the gender wage differential in the exporting firms is smaller than the one presented in the firms focused on the domestic market and if there are differences with respect to the level of development of the destination country of export, samples from all the twenty-six states of Brazil, in addition to the sample of the Federal District, were analyzed. The evidence pointed out that the exporting firms shows a larger gender wage-gap than the firms that sell only to the domestic market. Not even firms that export only to developed countries presented a decreased gender wage-gap. / Esta disserta??o apresenta dois ensaios acerca de rela??es do com?rcio internacional com o mercado de trabalho. No primeiro ensaio, o objetivo ? encontrar correla??es entre sal?rios m?dios de firmas, no Rio Grande do Sul, e o fato destas exportarem ou n?o os seus produtos. Buscou-se responder se os sal?rios m?dios das firmas exportadoras se diferenciam das que n?o participam do mercado internacional, se o n?vel de capital humano da firma tem influ?ncia sobre os poss?veis ganhos e se h? diferen?as com respeito ao pa?s de destino da exporta??o. Os resultados mostraram haver um pr?mio salarial para firmas que exportam para pa?ses desenvolvidos. Para os demais tipos de destino h? correla??o entre maiores n?veis de capital humano e o fato de a firma exportar para mercados com maior grau de competi??o. No segundo ensaio, analisa-se o impacto do com?rcio internacional no diferencial de sal?rios entre g?neros no Brasil. O objetivo ? buscar evid?ncias de contribui??o do fato de uma firma estar no mercado internacional (atrav?s da exporta??o) em amenizar a diferen?a salarial relacionada ao g?nero do trabalhador. Foram analisadas amostras separadas nos vinte e seis Estados do Brasil, al?m da amostra do Distrito Federal, e buscou-se responder se o diferencial de sal?rios nas firmas exportadoras ? menor do que o apresentado nas firmas voltadas ao mercado dom?stico. Ainda, verificou-se a possibilidade de haver diferen?as com respeito ao pa?s de destino da exporta??o. As evid?ncias apontaram que as firmas exportadoras apresentam diferencial de sal?rios ainda mais significativo do que as companhias voltadas para o mercado interno. Nem mesmo as firmas que exportam somente para pa?ses desenvolvidos apresentaram diferencial menos desfavor?vel ?s mulheres.
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Contribution à l'étude des mouvements de terrain de la région de Constantine (Algérie)Benaissa, Abdelkader 19 May 1989 (has links) (PDF)
Les formations argiIo~marneuses mio~pliocènes qui occupent de vastes étendues dans l'agglomération de Constantine, sont le siège de fréquentes manifestations d'instabilité, bien que la pente naturelle des versants soit faible (10 à 17 %). Dans cet ensemble argile-marneux, hétérogène à l'échelle métrique, la proportion de matériaux perméables est faible, mais certains niveaux de sables et de graviers ont une grande importance, car ils favorisent la mise en charge de l'eau dans le massif. Les informations apportées par les travaux de reconnaissance ainsi que la modélisation mécanique du phénomène sont concordantes et permettent de confirmer le rôle fondamental.
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新產品概念產生階段的市場知識來源之研究-以神達電腦公司之掌上型電腦/PDA的發展為例 / The Sources of Market Knowledge at the Stage of New Product Concept Generation曾齡瑩 Unknown Date (has links)
台灣高科技公司一直以來,在OEM/ODM領域有卓越的表現,隨著毛利逐步被壓縮之際,越來越多的我國廠商終於認知到,透過自有品牌(OBM)的建立是提升企業價值很重要的方式。但是,若要由OEM/ODM 跨入OBM,台灣廠商最缺乏的莫過於對「市場知識」的了解。本研究藉由對神達公司(近幾年內自有品牌Mio掌上型電腦/PDA上快速發展至全球前三大)的深入個案研究,探討「市場知識」的產生與能力建構之議題,並得到以下結論:
1、市場知識的來源似乎是很多元的;而對於剛採自有品牌策略之廠商的初期階段來說,似乎是較不完整的。
2、從事代工為主要業務型態的廠商,轉向發展自有品牌初期時,影響其市場知識來源的因素,受到內部因素(組織既有核心能力)影響及牽制的成分較多。
3、從事代工為主要業務型態的廠商,轉向發展自有品牌初期時,對其在取得完整市場知識的思考與運作上有所阻礙。儘管如此,組織既有核心能力對其所需要的部份市場知識仍有所助益。
4、企業在發展自有品牌的過程中,在取得市場知識與建構市場知識能力上似乎展現出「學習」的歷程。而在市場知識能力與行銷能力的建構上似乎又與高階主管的態度以及組織相關的管理有關。
5、除了本研究將「市場知識」定義為「關於顧客與競爭者的有系統、有意義且有用之知識」。從個案中發現,市場知識除了顧客與競爭者之外,其他環境因素也相當重要,例如政府的管制、社會文化的趨勢、科技的改變的趨勢等。
6、台灣資訊硬體製造能力,若能結合數位內容的創意,透過軟體與硬體的結合,似乎能創造更有價值和差異化的產品並創造更大的競爭優勢。 / Most of the high-tech corporations of Taiwan have remarkable performances in the OEM/ODM fields. Due to the compressing gross profit, more and more corporations realize that building own brand is a very important way to promote the enterprise value. However, if they want to make strides from OEM/ODM to OBM, the most deficient thing is the understanding of “Market Knowledge”. Through the case study of “MITAC international corp.”, we discuss the issue of the creation and the construction of ability of “Market Knowledge” and obtain the following conclusions.
1、The sources of “Market Knowledge” are diversified and they can be obtained by many different ways. For the new entered corporations in the OBM field, their sources of “Market Knowledge” are less complete at the initial period stage.
2、At the initial period stage of the OEM/ODM corporations in transformation to OBM, the factors which affect the sources of “Market Knowledge” are more affected and limited by the internal factors: the “Organizational Core Capabilities”.
3、At the initial period stage of the OEM/ODM corporations in transformation to OBM, they have the hindrance of the thinking and operation of obtaining the complete “Market Knowledge”. But, the “Organizational Core Capabilities” still can give some help in getting partial “Market Knowledge”.
4、In the process of obtaining “Market Knowledge” and establishing “Market Knowledge Capabilities”, there seems to be having some processes of “Learning”. And at the part of establishing “Market Knowledge Capabilities” and “Marketing Capabilities”, there seems to be having relationships with the senior managers’ attitudes and organizational management.
5、In this research, we define the meaning of “Market Knowledge” as the systematical, meaningful and useful knowledge which is about the customers and competitors. But from the case study, we find that “Market Knowledge” is not only about the customers and competitors, other environment reasons also have important impacts on it: the restrain of the government, the tendency of the society culture and the changes of technology.
6、If the OEM/ODM corporations can combine their hardware manufacturing capability with some contents of software, it seems a good way to them to create more values, more diversified products and more competition advantages.
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Morfogênese in vitro em tomateiro e berinjela e silenciamento gênico da sintase do mio-inositol-fosfato por RNAi em tomateiro / In vitro morphogenesis in eggplant and tomato plants and silencing of myo-inositolfosfate sintase gene by RNAi in tomato plantsFernandes, Denise 18 February 2009 (has links)
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Previous issue date: 2009-02-18 / Fundação de Amparo a Pesquisa do Estado de Minas Gerais / The main objectives in this work were: i) to find the optimum conditions to disinfect tomato seeds (Solanum lycopersicum Mill.) and eggplant seeds (Solanum melongena L.); ii) to evaluate the influence of the type of sealing in the obtained seedlings and explants; iii) to evaluate the effect of sonication in the morphogenesis in vitro of the tomato explants and in the viability of Agrobacterium tumefaciens cells; iv) to establish the parameters that allow the genetic transformation mediate by A. tumefaciens aiming the genetic silencing mediate by RNAi from myo-inositol-phosphate synthase, using the GmMPIS1 gene. It was checked that the use of deionized water was more efficient to disinfect eggplant seeds than 0.13% v/v chlorine solution. The use of dry disinfection in chlorine cameras is not appropriate to clean the tomato and eggplant seeds due to the gas toxicity and that it also compromises their germination. It was observed that gas exchange helps the seedlings development and leads to a bigger number of explants and with better quality to be used in the genetic transformation via Agrobacterium tumefaciens. Using the SAAT technique (Sonication-assisted Agrobacterium-mediated transformation), the explants and the bacteria suspensions were exposed to ultrasound for 0, 3, 6 and 9 seconds. It was verified that the immersion time from 3 to 6 seconds was appropriate to be used in genetic transformation, since it shown the biggest transient expression areas evaluated by in situ histochemical analysis of the GUS gene, the biggest number of regenerated structures and less mortality in the A. tumefaciens cells. The process was optimized when the immersion of the A. tumefaciens suspension was made 24 hours after the exposition to ultrasound. To make possible to select the transformed plants it was established the dependence of the hygromycin agent lethality and the found concentration of the non-transformed selected cells was 7.5 mg.L-1 in cotyledonary hipocotyledonary and leaf tomato explants. It was found that concentrations above this value were toxic, showing chlorotic and necrotic areas in the explants. A genetic transformation in tomato and eggplant plants was successfully made to check the relation between the MIPS gene and the seeds development by A. tumefaciens containing plasmids with silencing construction by siRNA to the MIPS gene using a conserved sequence of the soy gene GmMIPS. The transgenic nature of the primary regenerators was confirmed by in situ histochemical tests of GUS and by PCR analysis with specific oligonucleotides initiators. The analysis of the genetic expression confirmed the MIPS gene silencing and the morphologic analysis of the fruits confirmed the hypothesis of the relationship between the myo-inositol- phosphate synthase and the seeds development. However, as shown by flow-citometry technique, the process of regeneration in vitro used in the tomato plants transformation protocol changed some transgenic plants to polypoids. This was not observed in eggplant plants. / Este trabalho teve como objetivo: i) a otimização das condições de desinfestação de seentes de tomateiros (Solanum lycopersicum Mill.) e berinjela (Solanum melongena L.); ii) a avaliação da influência do tipo de vedação sobre a qualidade das plântulas e explantes oriundos destas; iii) a avaliação do efeito da sonicação sobre a morfogênese in vitro de explantes de tomateiros e sobre a viabilidade de células de Agrobacterium tumefaciens; iv) o estabelecimento de parâmetros para possibilitar a transformação genética mediada por A. tumefaciens visando ao silenciamento gênico mediado por RNAi da sintase do mio-inositol-fosfato, utilizando-se o gene GmMIPS1. Na desinfestação das sementes de berinjela, comprovou-se que tratamentos utilizando imersão em água deionizada são mais eficientes que imersão em solução de 0,13% v/v de cloro. A utilização de desinfestação a seco, em câmara de gás cloro, não é indicada para a assepsia de sementes de tomate e berinjela, pela toxicidade do gás às sementes, comprometendo sua germinação. Observou-se que as trocas gasosas favorecem o desenvolvimento das plântulas e geram explantes em maior número e de melhor qualidade para utilização de transformação genética via Agrobacterium tumefaciens. Ao se utilizar a técnica de SAAT ( Sonication-assisted Agrobacterium-mediated transformation ), os explantes e a suspensão bacteriana foram expostos a tempos de exposição ao ultra-som (0, 3, 6 e 9 segundos). Verificou-se que o intervalo de 3 a 6 segundos é o indicado para se utilizar em transformação genética, pois resultou nas maiores áreas de expressão transiente avaliada pela análise histoquímica in situ do gene GUS, maior número de estruturas regeneradas e menor mortalidade nas células de A. tumefaciens. O processo foi otimizado quando a imersão em suspensão de A. tumefaciens foi realizado após 24 horas de exposição ao ultra-som. Para ser possível a seleção de transformantes foi estabelecida a curva de letalidade ao agente higromicina e a concentração encontrada para seleção de células não transformadas foi de 7,5 mg.L-1 em explantes cotiledonares, hipocotiledonares e foliares de tomateiro. Dosagens acima de 7,5 mg.L-1 mostratam-se tóxicas, resultando em explantes com áreas cloróticas e necróticas. A fim de verificar a relação do gene MIPS com o desenvolvimento de sementes, a transformação genética foi realizada com sucesso em tomateiro e berinjela, via A. tumefaciens contendo plasmídeo com construção de silenciamento por siRNA para o gene MIPS, utilizando uma seqüência conservada do gene de soja GmMIPS. A natureza transgênica dos regenerantes primários foi confirmada mediante o teste histoquímico in situ de GUS e análise de PCR com oligonucleotídeos iniciadores específicos. A análise de expresão gênica confirmou o silenciamento do gene MIPS, e a análise morfológica dos frutos confirmou a hipótese do relacionamento da mio-inositol-fosfato-sintase com o desenvolvimento de sementes. Porém, conforme detectado pela técnica de citometria de fluxo, o processo de regeneração in vitro adotado no protocolo de transformação de tomateiro, ao contrário de berinjela, induziu poliploidia em algumas plantas transgênicas.
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