Les fermentations alcooliques peuvent s’accompagner de phénomènes de mortalité des levures entraînant des fermentations languissantes ou stoppées. Les mécanismes sous-jacents de la mortalité des levures en conditions de limitation nutritionnelle au cours des fermentations alcoolique sont encore mal connus. Dans ce travail, nous avons abordé la mortalité des levures en référence au schéma conceptuel développé dans les études de vieillissement cellulaire qui ont montré que la résistance à la carence peut être influencée par la nature du nutriment limitant la croissance cellulaire. Nous avons étudié l’apparition de la mort cellulaire en analysant la capacité des levures à mettre en place une réponse appropriée à différentes carences nutritionnelles. Nous avons montré que plusieurs carences nutritionnelles (acide oléique, ergostérol, acide pantothénique et acide nicotinique) entraînent une perte de viabilité de façon dépendante de l’azote. Nous avons démontré que la voie de signalisation azotée TOR/Sch9 est impliquée dans la mise en place de cette mort cellulaire. Dans de telles conditions, les levures n’acquièrent pas de résistance au stress du fait d’une modification à un niveau post-transcriptionnel. Nous avons examiné la capacité de différentes sources d’azote à entraîner la mort cellulaire, et nous avons montré qu’elles agissent différemment sur la mort cellulaire et que le NH4+ a une forte capacité à induire la mortalité. Enfin, les approches QTL nous ont permis d’identifier plusieurs régions contrôlant la mort cellulaire en limitation en acide oléique et acide pantothénique, cohérent avec un contrôle multigénique. 3 régions QTL communes à ces deux limitations ont été identifiées, ce qui suggère des mécanismes communs de contrôle de la survie des levures dans ces deux conditions de carences nutritionnelles. / Yeast cell death can occur during wine alcoholic fermentation and lead to sluggish or stuck fermentations. The mechanisms underlying cell death during yeast starvation in alcoholic fermentations remain unclear. In this work we addressed yeast cell death using conceptual framework from ageing studies showing that yeast resistance to starvation can be influenced by the nature of the nutrient limiting cell growth. We examined cell death occurrence considering yeast cells ability to elicit an appropriate response to a set of nutrient limitations. We showed that several micronutrients limitations (oleic acid, ergosterol, pantothenic acid and nicotinic acid) trigger cell death in a nitrogen-dependent manner. We provide evidence that the nitrogen Tor/Sch9 signaling pathway is involved in triggering cell death. In such conditions, yeast cells fail to acquire stress resistance given a restriction at a post-transcriptional level. We have examined the ability of different nitrogen sources to trigger cell death showing that they impact differentially on cell death and that NH4+ had a strong death inducing capacity. Finally, the QTL approaches allowed the mapping of a set of loci controlling cell death under oleic acid and pantothenic acid starvation that are consistent with a multigenic control. 3 QTL regions appeared to be common to these two limitations which suggests some common control of the yeast survival in these two nutrient-limited situations.

Orientador: Jose Barreto Campello Carvalheira / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-25T02:56:20Z (GMT). No. of bitstreams: 1 Mendes_MariaCarolinaSantos_D.pdf: 2658779 bytes, checksum: 153ed5344815e7e59a41c04c4a965670 (MD5) Previous issue date: 2014 / Resumo: A proteína mTOR é um proteína reguladora chave de vários processos celulares, dentre eles proliferação, crescimento e sobrevivência celular. Fatores de crescimento, oxigênio, status energético e a presença de aminoácidos são fundamentais para que todos esses processos ocorram normalmente. Descobertas realizadas nas últimas décadas mostraram que a via da mTOR encontra-se ativada em vários processos celulares, incluindo formação tumoral e angiogênese. A leucina é um aminoácido de cadeia ramificada que tem o maior potencial em ativar a via da mTOR. Devido sua capacidade de promover a síntese proteica e ganho de massa muscular, seu uso é constantemente estimulado em pacientes com câncer. No entanto, seus efeitos no crescimento tumoral não está claro. Dessa forma, realizamos um estudo cujo objetivo principal foi investigar os efeitos da dieta suplementada com leucina na modulação do crescimento tumoral em diferentes linhagens de células tumorais que se diferenciem em relação à ativação constitutiva da via IRS1/Akt/mTOR. Estudos in vivo e in vitro realizados demonstraram que as células que se diferenciam em relação à ativação da via IRS1/AKT/mTOR respondem de maneira distinta à suplementação com leucina. Linhagens de células tumorais que possuem a via da mTOR constitutivamente ativada, PC-3 e MCF-7, quando suplementadas com doses elevadas de leucina in vitro reduziram a proliferação celular e causaram retenção das células na fase G1 do ciclo celular. Já o xenoenxerto tumoral da PC-3 reduziu sua proliferação e aumentou a morte celular quando os animais foram suplementados com leucina na dieta. Nós também observamos aumento da atividade da mTOR e da p70S6K em todas as linhagens celulares quando suplementadas com leucina. O aumento da atividade da proteína mTOR foi acompanhado de redução na fosforilação de AKTser473 nas células que possuíam a via da PI3K hiperativada (PC-3 e MCF-7). Esse fato pode estar ocorrendo devido a ativação das alças de contraregulação ocasionadas pela estimulação excessiva provocada pela suplementação com leucina, naquelas linhagens celulares que já possuem a via hiperativada. Fato este comprovado pelo aumento da fosforilação em serina 307 da proteína IRS1. Dessa forma, nossos resultados sugerem que a ativação da via da mTOR é central para determinar a sensibilidade de tumores à dieta suplementada com leucina, podendo modular o desenvolvimento tumoral naquelas células que já possuem a via IRS1/AKT/mTOR constitutivamente ativada. O mecanismo pelo qual a leucina pode retardar o desenvolvimento tumoral em células que possuem a via da mTOR hiperativada parece estar relacionado com o eixo de regulação negativa p70S6K-PI3K, com consequente redução da fosforilação de AKT e liberação das vias apoptóticas nos tecidos tumorais / Abstract: mTOR is a key regulatory protein in various cellular processes including proliferation, cell growth and survival. Growth factors, oxygen, energy status and amino acids are all essential to these processes. New findings in the last few decades have shown that the mTOR pathway is activated in many cellular processes, including tumorigenesis and angiogenesis. The branched chain amino acid leucine has the greatest potential to activate the mTOR pathway. Due to its ability to promote protein synthesis and muscle mass gain, use of leucine is frequently utilized in patients with cancer. However, the effect of leucine on tumor growth is not clear. The aim of this study is therefore to investigate the effect of diet-supplemented leucine on the modulation of tumor growth in several tumor cell lines that differ in the constitutive activation status of the insulin receptor substrate 1 (IRS1)/AKT/mTOR pathway. Both in vitro and in vivo experiments demonstrated different cell proliferation responses when cells were exposed to high doses of leucine. Tumor cell lines PC-3 and MCF-7, which have a constitutively activated mTOR signaling, displayed reduced cell proliferation and G1 phase cell cycle arrest when supplemented with high doses of leucine in vitro. Likewise, leucine-supplemented PC-3 cell tumor xenografts displayed reduced proliferation and increased cell death. We also observed increased activity of mTOR and its downstream substrate p70S6K in all cell lines supplemented with leucine. Increased mTOR activity was accompanied by a reduction in AKT serine 473 (ser473) phosphorylation in cell lines with a hyperactivated PI3K pathway (PC-3 and MCF-7). This most likely occurred because leucine supplementation further increased mTOR and p70S6K activity, triggering the inhibitory p70S6K/IRS1 axis. In fact, we found increased IRS1 ser307 phosphorylation in hyperactivated cell lines (PC-3 and MCF-7) supplemented with high doses of leucine. Therefore, our results suggest that mTOR pathway activation is central to determining the sensitivity of tumors to leucine supplementation. Furthermore, this could affect the response to leucine-supplemented therapies of those tumors in which the PI3K pathway is constitutively activated. The mechanism for this appears to be related to the negative p70S6K/IRS1 regulation axis, with consequent reduction of AKT phosphorylation and the release of apoptotic pathways in tumor tissues / Doutorado / Fisiopatologia Médica / Doutora em Ciências

Ce travail de thèse avait pour objectif d’étudier la régulation de l’expression des gènes du métabolisme intermédiaire par les acides aminés alimentaires dans le foie de truite arc-en-ciel. Ces études ont permis de caractériser, pour la première fois dans le foie de truite, les principaux acteurs de la voie de signalisation Akt/TOR et leurs régulations. Nos résultats in vitro montrent qu’un mélange d’acides aminés, seul ou avec l’insuline, est capable de réguler l’expression de nombreux gènes impliqués dans la lipogenèse, la néoglucogenèse et la glycolyse. Les régulations observées en présence conjointe d’un mélange d’acides aminés et d’insuline semblent être, pour la plupart, dépendantes de la voie TOR. Par la suite, nous avons étudié l’effet de certains acides aminés comme la leucine (connue pour son effet « signal ») ainsi que la lysine et la méthionine (souvent ajoutées dans les aliments piscicoles riches en matières premières végétales afin d’atteindre l’équilibre en acides aminés). En présence d’insuline, la leucine, contrairement à la lysine et la méthionine, active la voie de signalisation TOR et régule l’expression de certains gènes (néoglucogenèse et lipogenèse) de façon similaire à un mélange d’acides aminés. Parallèlement, in vivo, nous avons étudié la régulation de l’expression des gènes du métabolisme intermédiaire lors d’un remplacement partiel ou total des huiles et farines de poisson par des produits végétaux dans l’aliment piscicole. Cette expérimentation a montré que, ni les voies de signalisation Akt/TOR, ni l’expression des gènes cibles ne sont affectés par ces nouveaux aliments. En conclusion, ces travaux ont montré que les acides aminés semblent jouer un rôle important dans la régulation de l’expression des gènes hépatiques du métabolisme intermédiaire chez la truite arc-en-ciel. / The objective of my PhD was to characterize the regulation of the intermediary metabolism related gene expression by dietary amino acids in the liver of rainbow trout. This work allowed us to characterize, for the first time in the liver of trout, the main proteins of the Akt/TOR signalling pathway and their regulations. In vitro results showed that a mixture of amino acids, in the presence or absence of insulin, is able to regulate the expression of numerous genes involved in lipolysis, gluconeogenesis and glycolysis. Such regulations induced by an amino acid mix together with insulin appear to be, at least partly, TOR-dependent. Afterwards, I studied the effect of specific amino acids known to be a signalling molecule (leucine) or having potential application as supplements to reach essential amino acid balance in plant ingredients-rich diet (lysine and methionine). In the presence of insulin, leucine, in contrast to lysine and methionine, is able to activate the TOR signalling pathway and regulate the expression of several genes involved in gluconeogenesis and lipogenesis in the same way as a mixture of amino acids. Furthermore, we studied in vivo, the effect of partial or total replacement of fish oil and fish meal by plant products in fish feed on .the regulation of intermediary metabolism related gene expression. This study showed that neither Akt/TOR signalling pathway nor the expression of the target genes were affected by such diets. In conclusion, these studies showed that amino acids seem to play an important role in the hepatic regulation of intermediary metabolism gene expression in the rainbow trout.

This thesis deals with contemporary approaches that provide higher protection of privacy of users. It focuses mainly on a group signature. In the practical part of this thesis I designed and implemented PS that is enable to gather information with the help of signal of a mobile device. The application was designed in accordance with fundamental cryptographic requirements such as the authenticity and the integrity of transmitted data. The anonymity of users is guaranteed through an application layer (the group signature) as well as through a network layer (Tor).

Vibrio cholerae is the cholera causing agent, divided into two biotypes, including the classical biotype and ElTor biotype. Both of these biotypes caused cholera epidemics in the world. The classical biotype caused 6th cholera pandemic (from 1921 to 1961), and ElTor biotype caused 7th cholera pandemic (from 1961 to the 70s). Haemolysin A, a hemolytic protein of V. cholerae ElTor biotype, is encoded by the hlyA gene. This gene is often used for analyzing genetic relationship between strains in the same species or between species in the same Vibrio genus. Results of analyzing nucleotide and amino acid sequences of hlyA gene of V. cholerae strain causing cholera in Vietnam (named hlyA.VN) showed that: the hlyA.VN gene sequence was similar to the hlyA gene sequences of V. cholerae strains of the 6thand 7thcholera epidemics. The hlyA gene of the 6th cholera epidemic strain was deficient in 11 nuleotides (this deficiency leading to the loss of 4 amino acids in the haemolysin A protein) comparing to hlyA.VN gene and hlyA gene of the 7th cholera epidemic strain. The results of genetic distance analysis as well as phylogenetic tree construction also confirmed V. cholerae causing cholera in Vietnam was closely relationship to the strains causing cholera pandemics in the world. It is great significance for the surveillance of molecular epidemiology to prevent cholera effectively. / Vibrio cholerae là tác nhân gây bệnh tả, được chia thành hai typ sinh học, đó là typ sinh học cổ điển và typ sinh học ElTor. Cả hai typ này đã từng gây ra các đại dịch tả trên thế giới. Typ sinh học cổ điển đã từng gây ra đại dịch tả lần thứ 6 (từ năm 1921 đến 1961), còn typ sinh học ElTor đã từng gây ra đại dịch tả lần thứ 7 (từ 1961 đến những năm 70). Haemolysin A, một protein có chức năng làm tan máu của V. cholerae typ sinh học ElTor, được mã hóa bởi gen hlyA. Gene này thường được sử dụng cho các phân tích quan hệ di truyền giữa các chủng trong cùng một loài V. cholerae hay giữa các loài trong cùng một chi Vibrio. Kết quả phân tích trình tự nucleotide và axit amin gen hlyA của chủng V. cholerae gâybệnh ở Việt Nam (hlyA.VN) cho thấy: trình tự gen hlyA.VN có sự tương đồng lớn với trình tự gen hlyA của chủng gây đại dịch tả 6 và 7. Gen hlyA của chủng gây đại dịch tả 6 bị thiếu hụt 11 nuleotide (sự thiếu hụt này dẫn tới sự mất đi 4 axit amin trong phân tử haemolysin A) so với gen hlyA.VN và gene hlyA của chủng gây đại dịch tả 7. Kết quả phân tích khoảng cách di truyền cũng như xây dựng cây phát sinh chủng loại cũng đã khẳng định: chủng gây bệnh ở Việt Nam có quan hệ rất gần với các chủng gây đại dịch tả trên thế giới. Nhận định này có ý nghĩa rất lớn đối với công tác giám sát dịch tễ học phân tử để ngăn chặn bệnh tả hiệu quả.

Numerous mobile applications have the potential to collect and share userspecific information on top of the essential data handling. This is made possible through poor application design and its improper implementation. The lack of security and privacy in an application is of main concern since the spread of sensitive and personal information can cause both physical and emotional harm, if it is being shared with unauthorized people. This thesis investigates how to confidentially transfer user information in such a way that the user remains anonymous and untraceable in a mobile application. In order to achieve that, the user will first authenticate itself to a third party, which provides the user with certificates or random generated tokens. The user can then use this as its communication credentials towards the server, which will be made through the Tor network. Further, when the connection is established, the WiFi details are sent periodically to the server without the user initiating the action. The results show that it is possible to establish connection, both with random tokens and certificates. The random tokens took less time to generate compared to the certificate, however the certificate took less time to verify, which balances off the whole performance of the system. Moreover, the results show that the implementation of Tor is working since it is possible for the system to hide the real IP address, and provide a random IP address instead. However, the communication is slower when Tor is used which is the cost for achieving anonymity and improving the privacy of the user. Conclusively, this thesis proves that combining proper implementation and good application design improves the security in the application thereby protecting the users’  privacy. / Många mobilapplikationer har möjlighet att samla in och dela användarspecifik information, utöver den väsentliga datahanteringen. Det här problemet möjliggörs genom dålig applikationsdesign och felaktig implementering. Bristen på säkerhet och integritet i en applikation är därför kritisk, eftersom spridning av känslig och personlig information kan orsaka både fysisk och emotionell skada, om den delas med obehöriga personer. Denna avhandling undersöker hur man konfidentiellt kan överföra användarinformation på ett sätt som tillåter användaren av mobilapplikationen att förbli både anonym och icke spårbar. För att uppnå detta kommer användaren först att behöva autentisera sig till en tredje part, vilket förser användaren med slumpmässigt genererade tecken eller med ett certifikat. Användaren kan sedan använda dessa till att kommunicera med servern, vilket kommer att göras över ett Tor-nätverk. Slutligen när anslutningen upprättats, kommer WiFi-detaljerna att skickas över periodvis till servern, detta sker automatiskt utan att användaren initierar överföringen. Resultatet visar att det är möjligt att skapa en anslutning både med ett certifikat eller med slumpmässiga tecken. Att generera de slumpmässiga tecknen tog mindre tid jämfört med certifikaten, däremot tog certifikaten mindre tid att verifiera än tecknen. Detta resulterade i att de båda metoderna hade en jämn prestanda om man ser över hela systemet. Resultatet visar vidare att det implementeringen av Tor fungerar då det är möjligt för systemet att dölja den verkliga IPadressen och att istället tillhandahålla en slumpmässig IP-adress. Kommunikationen genom Tor gör dock systemet långsammare, vilket är kostnaden för att förbättra användarens integritet och uppnå anonymitet. Sammanfattningsvis visar denna avhandling att genom att kombinera korrekt implementering och bra applikationsdesign kan man förbättra säkerheten i applikationen och därmed skydda användarnas integritet.

An integrative, biochemical, genetic, and molecular biology approach utilizing gene manipulation, gene knock outs, plasmid based protein expression, and in vivo protein localization of fluorescence tagged proteins was employed to determine the function of an essential protein, Lst8, in TORC1 and TORC2 signaling and a previously uncharacterized complex, the Far3-7-8-9-10-11 complex (Far complex) in the budding yeast, Saccharomyces cerevisiae. Mutations in SAC7 and FAR11 suppressed lethality of both lst8 and tor2-21 mutations but not TORC1 inactivation, suggesting that the essential function of Lst8 is linked only to TORC2. Far11, a component of a six-member complex, was found to interact with Tpd3 and Pph21, conserved components of Protein Phosphatase 2A (PP2A) via co-immunoprecipitation. Mutations in FAR11 and RTS1, which encodes a PP2A regulatory B subunit, restore phosphorylation to the TORC2 substrate Slm1 in a tor2-21 mutant. These data suggest that TORC2 signaling is antagonized by Far11-dependent PP2A activity. To characterize the assembly of the Far complex in vivo, intracellular localization of the Far complex was examined by fluorescence microscopy. It was found that the Far complex localizes to the endoplasmic reticulum (ER). The data show that Far9 and Far10 are tail-anchored proteins that localize to the ER first and recruit a Far8-Far7-Far3 pre-complex. Far11 is found at the ER only when all other Far proteins are assembled at the ER. Surprisingly, ER localization is required for the Far Complex’s role TORC2 signaling because deletion of the tail-anchor domain of Far9 results in partial bypass of the tor2-21 mutant growth defect at 37 ˚C.

Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the third most common cause of cancer-related deaths. Systemic therapies are the main treatment options for HCC patients with advanced disease (∼ 80% of all cases). However, only very moderate clinical responses are achieved with most of the conventional therapies. Thus, more effective therapeutic strategies are much needed. The PI3K/Akt/mTOR signaling pathway, which plays a critical role in controlling cell proliferation and survival, is aberrantly activated in ∼ 45% HCC, suggesting it to be a potential target for HCC treatment. Moreover, emerging evidences indicate that activation of the PI3K/Akt/mTOR pathway may be associated with resistance to many cytotoxic chemotherapies, including microtubule targeting agents. In this study, by gene expression profiling and gene ontology analysis, "microtubule-related cellular assembly" was identified to be the major biological/functional process involved in HCC development, suggesting that microtubule is also an important therapeutic target for HCC. With these understandings, it is hypothesize in this thesis that combined targeting of a key component ofthe PI3K/Akt/mTOR pathway, namely the mammalian target of rapamycin (mTOR) and the microtubule would be an effective therapeutic strategy for HCC. The objectives of the thesis are to examine the therapeutic potential of microtubule targeting, mTOR targeting, and combined targeting of the microtubule and mTOR in both in vitro and in vivo models of HCC. / In summary, the PI3K/Akt/mTOR pathway and the microtubule represent promising therapeutic targets for HCC treatment. The findings from this thesis offer a rationale for combining mTOR inhibitors with microtubule targeting agents for effective HCC treatment. / In the second part, the effect of mTOR inhibition, either alone or in combination with an additional microtubule targeting agent (vinblastine) was investigated in HCC. Temsirolimus, an mTOR inhibitor, suppressed HCC cell proliferation in as early as 24 hrs with an IC50 of 1.27+/-0.06muM (Huh7), 8.77+/-0.76muM (HepG2), and 52.95+/-17.14muM (Hep3B). Vinblastine (1nM) alone caused 30--50% growth inhibition in 3 HCC cell lines. In these HCC cell lines, it was found that temsirolimus/vinblastine combination resulted in an additive to synergistic effect (when compared to single agents alone) with maximum growth inhibition of 80--90% as early as 24 hrs upon treatment. This marked growth inhibition was accompanied with cell cycle arrest at both G1 and G2/M phases, and PARP cleavage (a hallmark for apoptosis). Moreover, the combination specifically caused concerted down-regulation of several important anti-apoptotic and survival proteins (survivin, Bcl-2 and Mcl-1), which was not observed in single agent treatments. It was hypothesized that inhibition of these key anti-apoptotic/survival proteins may represent a novel mechanistic action of this highly effective combination approach of dual targeting of mTOR and microtubule by temsirolimus/vinblastine in HCC cells. Indeed, transient over-expression of each of these genes (survivin, Bcl-2 or Mcl-1) in HCC cells did partially rescue the growth inhibitory effect of the temsirolimus/vinblastine combination. More importantly, this novel combination significantly suppressed the growth of HCC xenografts in nude mice (when compared with single agents alone). / In the third part, the anti-tumor effect of another mTOR inhibitor everolimus in combination with microtubule targeting agents, vinblastine and patupilone (a microtubule-stabilizing agent), was investigated in HCC cells. Everolimus/vinblastine combination resulted in an additive to synergistic effect accompanied with cell cycle arrest at both G1 and G2/M phases, and PARP cleavage. The combination also caused concerted down-regulation of anti-apoptotic and survival proteins (survivin, Bel-2 and Mel-1) as observed with the temsirolimus/vinblastine combination. However, everolimus only moderately enhanced the sensitivity of patupilone for reasons unknown. / Taxanes are the major chemotherapeutic agents that target the microtubule. In the first part of the thesis, the anti-tumor activity of two taxanes, paclitaxel and docetaxel (which are known to stabilize microtubules) was examined and compared with doxorubicin (a DNA intercalating agent). Across all three HCC cell lines tested, it was found that the microtubule targeting agents, taxanes, were more efficacious than doxorubicin. This supports the initial finding that microtubule assembly process is functionally important in HCC. Recent studies demonstrated that using nanoparticles for drug delivery can greatly enhance therapeutic efficacy and reduce side-effects. Therefore, the nanoparticle albumin-bound (nab)-paclitaxel was employed to further evaluate the therapeutic efficacy of such a delivery strategy in HCC models. In all three HCC cell lines tested, nab-paclitaxel was found to be the most effective agent, with an average IC50 value of 0.16--10.42nM, when compared to non-conjugated taxanes (paclitaxel, docetaxel) and doxorubicin. In vitro analysis showed that nab-paclitaxel was able to induce cell cycle arrest at G2/M phase and apoptosis in HCC cells. In vivo study demonstrated that nab-paclitaxel readily inhibited the growth of HCC xenografts with lower toxicity when compared to paclitaxel, docetaxel and doxorubicin. Moreover, specific silencing of a key regulatory protein for microtubule dynamics, Stathmin 1, by siRNA significantly enhanced the effect of nab-paclitaxel in HCC cells, resulting in synergistic growth inhibition in vitro. / Zhou, Qian. / Advisers: Winnie Yeo; Vivian Lui; Nathalie Wong. / Source: Dissertation Abstracts International, Volume: 73-06, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 148-164). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Dissertação (mestrado)—UnB/UFPB/UFRN, Programa Multi-institucional e Inter-regional de Pós-graduação em Ciências Contábeis, 2016. / Esta dissertação teve por objetivo analisar o efeito da incerteza sobre o nível de investimento das empresas listadas na Bolsa de Valores, Mercadorias e Futuros de São Paulo (BM&FBovespa). A amostra compôs-se de todas as empresas não financeiras listadas na BM&FBovespa entre o período de 2008 a 2014. Coletaram-se os dados contábeis e de mercado no banco de dados Economatica®, e fez-se a classificação dos setores das empresas de acordo com os dados disponibilizados na BM&FBovespa. Como proxy para a incerteza total, utilizou-se a volatilidade histórica do retorno das ações. Decompôs-se essa medida em três componentes de risco: o de mercado, o idiossincrático e o específico da firma, baseando-se no modelo de Bulan (2005) e Xu, Wang e Xin (2010). Além disso, utilizou-se uma proxy alternativa para medição da incerteza, conforme o estudo de Tran (2014), que foi a volatilidade condicional do retorno das ações. Para análise do efeito da incerteza sobre as decisões de investimento, utilizou-se o método de Mínimos Quadrados Ordinários (MQO) e Mínimos Quadrados em Dois Estágios (MQ2E), quando constatados problemas de endogeneidade da medida de incerteza. Os resultados indicam que a incerteza total tem um efeito negativo e significativo estatisticamente sobre o nível de investimento. Quanto às incertezas de mercado e idiossincrática, verificou-se que não há um efeito estatisticamente significativo sobre o nível de investimento, enquanto que a incerteza específica tem um efeito negativo e significativo, inclusive superior ao da incerteza total, uma vez que o aumento de 1% da incerteza específica da firma reduz o nível de investimento em 0,4706%, e o aumento de 1% da incerteza total reduz o nível de investimento em apenas 0,2567%. Assim, pode-se afirmar que a incerteza da firma é mais significante para explicar o nível de investimento de uma empresa do que a incerteza total, logo, a abordagem que explica o efeito da incerteza sobre o nível de investimento é a Teoria das Opções Reais (TOR). Diante dessas evidências, rejeitaram-se a hipótese 1, de que a incerteza de mercado tem um efeito negativo sobre o nível de investimento, e a hipótese 2 (a), de que a incerteza idiossincrática tem um efeito negativo sobre o nível de investimento. Por outro lado, não se pode rejeitar a hipótese 2, de que existe um efeito negativo da incerteza total sobre o nível de investimento, e a hipótese 2 (b), de que existe um efeito negativo da incerteza específica da firma sobre o nível de investimento. Além disso, de acordo com os resultados obtidos, verificou-se que, no mercado brasileiro, as firmas menores são mais sensíveis à incerteza e às mudanças no ambiente de negócio do que as firmas maiores. Quanto à competitividade, identificou-se que as empresas mais competitivas são mais sensíveis à incerteza do que as menos competitivas. Ademais, a amostra do estudo se restringiu ao período de full IFRS, tendo em vista as mudanças das práticas contábeis que alteraram a forma de mensuração de algumas variáveis explicativas. Dessa forma, após essa restrição, verificou-se que não houve variação do efeito das incertezas total, de mercado, de setor e específica da firma sobre o nível de investimento. Nesse sentido, com base nos resultados obtidos, observou-se que a incerteza tem um efeito negativo sobre o nível de investimento das empresas, principalmente a incerteza específica da firma, conforme prevê a TOR. Logo, os administradores devem considerá-la no processo de tomada de decisão. Ressalta-se que esses resultados podem fomentar o estabelecimento de políticas para análise da viabilidade de novos projetos de investimento. Por fim, as conclusões obtidas nesta dissertação devem ficar restritas à amostra utilizada. / This dissertation aimed to analyze the effect of uncertainty on the level of investment by companies listed on the São Paulo Stock, Mercantile and Futures Exchange (BM&FBovespa). The sample consists of all non-financial companies listed on the BM & FBovespa between 2008 and 2014. They were collected accounting and market data in Economatica®database, and the classification of sectors of the companies was made according to the data available on the BM&FBovespa. As a proxy for the total uncertainty, it was used the historical volatility of stock returns. This measure was decomposed into three risk components: market, idiosyncratic and specific to the firm, based on the model of Bulan (2005) and Xu, Wang Xin (2010). In addition, it was used an alternative proxy to measurement the uncertainty, according to the study of Tran (2014), which was the conditional volatility of the stock return. To analyze the effect of uncertainty on investment decisions, it was used the method of Ordinary Least Squares (OLS) and Two-Stage Least Squares (2SLS), when observed endogeneity problems of measurement of uncertainty. The results indicate that the total uncertainty has a statistically negative and significant effect on the level of investment. Regarding the market and idiosyncratic uncertainties, it was found there is no statistically significant effect on the level of investment, whereas the specific uncertainty has a significant negative effect, even higher than the total uncertainty, since the increase of 1 % of the specific firm uncertainty reduces the level of investment in 0.4706%, and the increase of 1% of the total uncertainty reduces the level of investment in just 0.2567%. So it can say that the uncertainty of the firm is more important to explain the company’s investment level than the total uncertainty, so the approach that explains the effect of uncertainty on the level of investment is the Real Options Theory (ROT). Faced with this evidence, they rejected the hypothesis 1, that market uncertainty has a negative effect on the level of investment, and the hypothesis 2 (a), that the idiosyncratic uncertainty has a negative effect on the level of investment. Furthermore, according to the results, it was found that, in Brazilian market, smaller firms are more sensitive to uncertainties and changes in the business environment than the bigger firms. As for competitiveness, it was identified that the most competitive companies are more sensitive to uncertainty than the less competitive. Moreover, the study sample was restricted to the period of full IFRS, considering the changes in accounting practices that changed the way of measurement of some explanatory variables. Therefore, after this restriction, it was found that there was no variation of the effect of total market, sector, and specific of firm uncertainties on the level of investment. In this sense, based on the results, it was observed that the uncertainty has a negative effect on the level of business investment, particularly the specific uncertainty of the firm, as predicts the TOR. Therefore, administrators should consider it in the decision-making process. It is noteworthy that these results may encourage the establishment of policies for viability analysis of new investment projects. Finally, the conclusions obtained in this dissertation should be restricted to the sample used.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-12-28T17:28:07Z No. of bitstreams: 1 476880 - Texto Completo.pdf: 599451 bytes, checksum: cd9bc0981219eb2d393eed6f52e56c65 (MD5) / Made available in DSpace on 2015-12-28T17:28:07Z (GMT). No. of bitstreams: 1 476880 - Texto Completo.pdf: 599451 bytes, checksum: cd9bc0981219eb2d393eed6f52e56c65 (MD5) Previous issue date: 2015-03-27 / Objectives: To identify the variables related to morbidity and risk factors for death of patients that undergo to urgent myocardial revascularization surgery. Method: Historical cross sectional study. It included 232 patients aged ?18 years old that underwent to isolated myocardial revascularization surgery, with or without cardiopulmonary bypass. Those surgeries were urgents and were adopted as emergency measures. It happened from January 1996 to December 2014. The total of revascularization myocardial surgeries carried out in this period were 3772 Results: The frequency of death observed in this sample was 117 (50.4%). Multivariate analysis identified as risk predictors: age ?65 years old (OR: 2.34; 95% CI 1.35 to 5.27; P = 0.007), congestive heart failure class III and IV (OR: 2, 67, 95% CI 1.35 to 5.28; P = 0.005), cardiopulmonary bypass time> 90 minutes (OR: 2.52, 95% CI 1.21 to 4.17; P = 0.001), cardiogenic shock (OR: 3.47; 95% CI 1.70 to 7.07; P = 0.001), acute myocardial infarction (OR: 2.73, 95% CI 1.25 to 5.94, P = 0.011), bleeding (OR: 3.06, 95% CI 1.00 to 9.37; P = 0.049). Conclusion: Age, chronic obstructive pulmonary disease, class III and IV of congestive heart failure, cardiopulmonary bypass time, cardiogenic shock, acute myocardial infarction and bleeding they were independent predictors of in-hospital mortality in patients that undergo to myocardial revascularization surgery. / Objetivos: identificar as vari?veis relacionadas ? morbidade e os fatores de risco para o ?bito de pacientes submetidos ? revasculariza??o do mioc?rdio de urg?ncia e emerg?ncia. M?todos: Estudo de coorte hist?rica. Inclu?dos 232 pacientes com idade ?18 anos, submetidos ? cirurgia de revasculariza??o do mioc?rdio isolada, com ou sem uso de circula??o extracorp?rea, em car?ter de urg?ncia e emerg?ncia, de janeiro de 1996 a dezembro de 2014, de um total de 3772 cirurgias de revasculariza??o do mioc?rdio realizadas neste per?odo. Resultados: A frequ?ncia de ?bito observada nesta amostra foi de 50,4% (117 pacientes). A an?lise multivariada identificou como preditores de risco para mortalidade: idade maior que 65 anos (OR: 2,35; IC 95% 1,27?4,35; P= 0,007), insufici?ncia card?aca congestiva classe III e IV (OR: 2,67, IC 95% 1,36?5,28; P= 0,005), doen?a pulmonar obstrutiva cr?nica (OR: 2,49; IC 95% 1,21?5,11; P=0,013) e tempo de circula??o extracorp?rea maior que 90 minutos (OR: 2,52, IC 95% 1,22?4,17; P= 0,010). Conclus?o: Idade, doen?a pulmonar obstrutiva cr?nica, classe III e IV da insufici?ncia card?aca congestiva, tempo de circula??o extracorp?rea, foram preditores independentes de mortalidade intra-hospitalar de pacientes submetidos ? cirurgia de revasculariza??o do mioc?rdio de urg?ncia e de emerg?ncia.