Molecular mechanisms of prostatic carcinoma growth and progression

Prostate cancer represents the most commonly diagnosed malignancies in American men and is the second leading cause of male cancer deaths. The overall objectives of this research were designed to understand the cellular and molecular mechanisms of prostatic carcinoma growth and progression. This dissertation was divided into two major parts: (1) to clone and characterize soluble factor(s) associated with bone that may mediate prostatic carcinoma growth and progression; (2) to investigate the roles of extracellular matrix in prostatic carcinogenesis.^ The propensity of prostate cancer cells to metastasize to the axial skeleton and the subsequent osteoblastic reactions observed in the bone indicate the possible reciprocal cellular interaction between prostate cancer cells and the bone microenvironment. To understand the molecular and cellular basis of this interaction, I focused on the identification and cloning of soluble factor(s) from bone stromal cells that may exert direct mitogenic action on cultured prostate cells. A novel BPGF-1 gene expressed specifically by bone and male accessory sex organs (prostate, seminal vesicles, and coagulating gland) was identified and cloned.^ The BPGF-1 was identified and cloned from a cDNA expression library prepared from a human bone stromal cell line, MS. The conditioned medium (CM) of this cell line contains mitogenic materials that induce human prostate cancer cell growth both in vivo and in vitro. The cDNA expression library was screened by an antibody prepared against the mitogenic fraction of the CM.^ The cloned BPGF-1 cDNA comprises 3171 nucleotides with a single open reading frame of 1620 nucleotides encoding 540 amino acids. The BPGF-1 gene encodes two transcripts (3.3 and 2.5 kb) with approximately equal intensity in human cells and tissues, but only one transcript (2.5 kb) in rat and mouse tissues. Southern blot analysis of human genomic DNA revealed a single BPGF-1 gene. The BPGF-1 gene is expressed predominantly in bone and seminal vesicles, but at a substantially lower level in prostate. Polyclonal antibodies generated from synthetic peptides that correspond to the nucleotide sequences of the cloned BPGF-1 cDNA reacted with a putative BPGF-1 protein with an apparent molecular weight of 70 kDa. The conditioned media isolated from COS cells transfected with BPGF-1 cDNA stimulated the proliferation and increased the anchorage-independent growth of prostate epithelial cells. These findings led us to hypothesize that BPGF-1 expression in relevant organs, such as prostate, seminal vesicles, and bone, may lead to local prostate cancer growth, metastasis to the seminal vesicles, and subsequently dissemination to the skeleton.^ To assess the importance of extracellular matrix in prostatic carcinogenesis, the role of extracellular matrix in induction of rat prostatic carcinoma growth in vivo was evaluated. NbE-1, a nontumorigenic rat prostatic epithelial cell line, was induced to form carcinoma in athymic nude hosts by coinjecting them with Matrigel and selected extracellular matrix components. Induction of prostatic tumor formation by laminin and collagen IV was inhibited by their respective antibodies. Prostatic epithelial cells cloned from the tumor tissues were found to form tumors in athymic nude hosts in the absence of exogenously added extracellular matrix. These results suggest that extracellular matrix induce irreversibly prostatic epithelial cells that behave distinctively different from the parental prostatic epithelial cell line. ^

Modelo multidimensional de los factores que afectan la interacción entre marketing y ventas en el lanzamiento de nuevos productos en Latino América

Esta tesis se centra en la identificación y análisis de los factores que pueden favorecer o actuar como barreras del éxito de la implementación de la innovación y las relaciones entre sí, desde el enfoque de la interface marketing-ventas. El trabajo empírico se enmarca en el vacío de investigación existente en el campo del proceso de lanzamiento de nuevos productos en los mercados donde operan subsidiarias de empresas multinacionales de consumo masivo (FMCG). Las empresas FMCG son altamente dependientes de la innovación como proceso clave determinante del crecimiento competitivo de mediano y largo plazo. En un contexto de acortamiento del ciclo de vida de los productos, como resultado del desarrollo tecnológico y científico que impactan en el comportamiento de los consumidores, las empresas invierten un mayor nivel de recursos en el desarrollo de nuevos productos, reingeniería y programas de innovación (Mundra, Gulati y Gupta, 2013). Sin embargo, a pesar del aumento en la inversión, las tasas de éxito de la innovación reportadas son inferiores al 25% (Evanschitzky, Eisend, Calantone y Jiang, 2012). Aumentar las tasas de éxito de los proyectos de innovación es reconocida en la literatura como un elemento clave para la supervivencia y competitividad de las empresas, para ser superiores a su competencia y desarrollar nuevos modelos de negocios. A pesar de la existencia de estudios que intentan comprender el proceso de lanzamiento de nuevos productos, no se ha identificado un claro prototipo de gestión de la innovación (Gupta et al, 2007). Profundizando en los factores de éxito, los autores Keupp, Palmié y Gassman (2012) reconocen que la innovación exitosa no depende solamente de la estrategia de selección de los proyectos de innovación, sino también la forma en que los mismos son implementados (Klein and Sorra, 1996; Repenning, 2002; Keupp, Palmié y Gassmann, 2012). Al analizar la implementación de los proyectos de lanzamiento de nuevos productos al mercado, en empresas FMCG, dicho proceso es responsabilidad principalmente de las funciones de marketing y ventas a través de la comunicación con los consumidores y los clientes respectivamente (Ernst, Hoyer y Rubsaamen, 2010). Es decir que el éxito en la implementación de la innovación requiere la gestión efectiva de la relación inter-funcional entre marketing y ventas (Ernst, Hoyer y Rubsaamen, 2010; Hughes, Le Bon y Malshe, 2012). A pesar de la importancia de la integración entre marketing y ventas en la conceptualización e implementación de la innovación, este tema no ha sido estudiado en profundidad (Hughes, Le Bon y Malshe, 2012; Keupp, Palmié y Gassmann, 2012). En las empresas multinacionales, está demostrado que el desempeño de las subsidiarias determinan el éxito competitivo de la empresa a nivel global. El desafío de dichas subsidiarias es conjugar el desarrollo global de innovación y comunicación con las características locales de comportamiento del consumidor y el mercado. Por lo tanto, esta investigación empírica responde a la pregunta académica y de gestión acerca de cómo mejorar las tasas de éxito de lanzamiento de nuevos productos al mercado en subsidiarias de empresas de consumo masivo, desde la perspectiva de la relación entre marketing y ventas. En particular analiza cómo afectan la formalización de los procesos y los mecanismos de comunicación a la confianza interpersonal y a la efectividad de la interface marketing-ventas y dichos factores a su vez sobre la planificación integrada de la implementación de la innovación. La determinación de los factores o ítems que conforman cada uno de los constructos del proceso de ejecución de la innovación, se llevó a cabo a partir de una revisión exhaustiva del estado del arte de la literatura sobre las interfaces funcionales y el proceso de innovación. Posteriormente, los ítems seleccionados (más de 50 en total) fueron validados por referentes de marketing y ventas de Argentina y Uruguay a través de entrevistas en profundidad. A partir de los factores identificados se construyeron dos modelos teóricos: • (1) relativo a la influencia de las dimensiones de confianza interpersonal sobre la efectividad de las uniones inter-funcionales y como los mecanismos organizacionales, tales como la frecuencia y la calidad de la comunicación entre las áreas, afectan la confianza y la relación entre ellas; • (2) relativo a la dimensión planificación integrada de la implementación de la innovación, ya que durante el lanzamiento de nuevos productos al mercado, marketing y ventas utilizan procesos formales que facilitan la comunicación frecuente y efectiva, desarrollando confianza inter-personal que no solamente afecta la efectividad de su relación sino también el desarrollo de planes integrados entre ambas áreas. El estudio fue llevado a cabo en una empresa multinacional de consumo masivo que integra la lista Global 500 (Fortune, 2015), presente en todo el mundo con más de 25 marcas participantes en más de 15 categorías, implementando 150 proyectos de innovación en el último año. El grupo de subsidiarias en estudio fue reconocido a nivel mundial por su desempeño en crecimiento competitivo y su alta contribución al crecimiento total. El modelo analizado en esta tesis fue expandido al resto de América Latina, tratándose entonces de un caso ejemplar que representa una práctica de excelencia en la implementación de la innovación en subsidiarias de una empresa multinacional. La recolección de los datos fue llevado a cabo a través de un cuestionario estructurado y confidencial, enviado a la base de datos de todo el universo de directores y gerentes de marketing y ventas. El nivel de respuesta fue muy elevado (70%), logrando 152 casos válidos. El análisis de datos comprendió el análisis descriptivo de los mismos, estimación de fiabilidad y análisis factorial exploratorio a través del software SPSS v.20. El análisis factorial confirmatorio y el análisis de senderos para examinar las relaciones entre los factores se estudiaron mediante el software R (Package 2.15.1., R Core Team, 2012) (Fox, 2006). Finalmente se llevaron a cabo entrevistas en profundidad a gerentes de marketing y ventas de cada uno de los seis países con el fin de profundizar en los constructos y sus relaciones. Los resultados de los modelos demuestran que la frecuencia de comunicación impacta positivamente en la calidad de la misma, que a su vez afecta directamente la relación entre marketing y ventas. Adicionalmente, la calidad de la comunicación impacta sobre la confianza cognitiva, que a su vez se relaciona no solamente con la confianza afectiva sino también con la relación entre ambas áreas. Esto significa que para mejorar la implementación de la innovación, los gerentes deberían orientarse a reforzar la relación entre marketing y ventas facilitando la construcción de confianza interpersonal primero cognitiva y luego afectiva, incrementando la frecuencia de la comunicación que alimenta la calidad de la comunicación entre ambas áreas. A través del segundo modelo se demuestra que durante el lanzamiento de nuevos productos al mercado, marketing y ventas necesitan emplear procesos formales que faciliten la comunicación frecuente y efectiva. De esta forma se contrarresta el efecto negativo de la formalización sobre la planificación integrada entre ambas áreas. Adicionalmente, los gerentes de ambos departamentos deberían promover la construcción de confianza interpersonal, no solamente para mejorar la efectividad de la relación, sino también para desarrollar planes integrados de implementación de nuevos productos. Finalmente, se valida que la frecuencia de la comunicación, la confianza afectiva y la relación marketing-ventas, se relacionan positivamente con la planificación integrada en la implementación de la innovación. El estudio contribuye a la comprensión de los factores que las empresas pueden emplear para mejorar la relación inter-funcional entre marketing y ventas y la implementación de la innovación en empresas de consumo masivo. El aporte de esta investigación puede ser valorado de dos maneras, los aportes a la gestión y a la academia. Desde el punto de vista empresarial, provee a los líderes al frente de empresas de consumo masivo, del conocimiento sobre los factores que afectan la implementación de la innovación y en definitiva el éxito del negocio a mediano y largo plazo. Desde el punto de vista académico aporta al conocimiento del proceso de implementación de la innovación y en la efectividad de la interface marketing y ventas en un caso de buenas prácticas en el mercado de consumo masivo. A su vez incorpora por primera vez un estudio empírico en geografías emergentes capaces de recuperar el camino de crecimiento posterior a una profunda crisis económica a través de la exitosa implementación de la innovación en sus mercados. ABSTRACT This thesis is focused on the identification, analysis and relationship study of factors which may benefit or hinder the successful deployment of innovation, from a marketing-sales interface perspective. Considering the non-existent investigation dedicated to the study of new products launches into markets in which Fast Moving Consumer Goods (FMCG) companies’ subsidiaries operate, it is that this investigation has been carried out. FMCG companies rely on innovation as their key process for a competitive growth on a medium and long term basis. Nowadays, the life-cycle of products is getting shorter as a result of new technological and scientific development, having impact on consumer behavior, and therefore companies are forced to dedicating more resources to the development of new products, reengineering and innovation programs (Mundra, Gulati and Gupta, 2013). However, in spite of the investment increase, the innovation success rates have been reported to be lower than 25% (Evanschitzky, Eisend, Calantone y Jiang, 2012). Increasing success rates on innovation processes has been considered as a key element on the survival and competitiveness of companies, outperforming competitors and developing new business models. Despite new studies which try to comprehend the process of new products launch, a prototype of innovation management has not yet been identified (Gupta et al, 2007). Emphasizing on success factors, authors Keupp, Palmié and Gassman (2012) recognize that successful innovation does not solely depend on innovation processes’ selection strategy, but it is also based on the way in which these are implemented (Klein and Sorra, 1996; Repenning, 2002; Keupp, Palmié y Gassmann, 2012). While analyzing the implementation of projects for new products releases on massive consumption companies, the two departments in charge of taking this forward are marketing and sales, by focusing on communication strategies with consumers and clients respectively (Ernst, Hoyer y Rubsaamen, 2010). This means that having success on innovation implementation requires an effective management of inter-functional relationship among marketing and sales (Ernst, Hoyer y Rubsaamen, 2010; Hughes, Le Bon y Malshe, 2012). In spite of the importance on the integration between marketing and sales on the conceptualization and implementation of innovation, this subject has not been studied in depth (Hughes, Le Bon y Malshe, 2012; Keupp, Palmié y Gassmann, 2012). In multinational companies, previous research has confirmed that the performance of their subsidiaries determine the competitive success of the company on a global scale. The challenge of said subsidiaries is to conjugate the global innovation development and communication with the local consumer and market behavior. Therefore, this empirical study aims to respond to the academic and management question of how to improve the success rates of new product launches into MNE subsidiary’ markets, from a marketing-sales relationship perspective. Particularly, this investigation analyses how the formalization of products and communication mechanisms affect interpersonal trust and marketing-sales interface effectiveness and also on how these factors affect the overall planning of the implementation of innovation. The determination of which factors build the hypothesis of the innovation execution process was taken forward through an extensive research on the extant literature on functional interfaces and innovation processes. More than 50 items were selected which were in turn validated by marketing and sales referents on Uruguay and Argentina through in depth interviews. Based on the identified factors, two theoretical models were proposed: (1) Relative to the influence that interpersonal trust dimensions have on inter functional linkages effectiveness and how organizational mechanisms such as frequency and quality of communication between departments affect trust and their relationship. (2) Relative to the integrated planning and innovation implementation dimensions. Marketing and sales department use formal process thus allowing inter-personal trust, which affects positively their relationship and also enables the development of integrated planning between them. The study was performed within a massive consumption company which is part of the “Global 500” (Fortune, 2015), with subsidiaries all over the world and more than 25 participant brands in 15 categories, having implemented over 150 innovation projects in the year under study. The analyzed subsidiary group has been awarded worldwide for their performance in competitive growth and their high contribution to the total growth. The model being analyzed in this thesis was implemented throughout Latin America, representing a remarkable case of innovation implementation excellence for subsidiaries of multinational companies. Data recollection was carried out through a structured and confidential questionnaire, sent to the universe of marketing-sales directors and managers’ database available with a high level of responsiveness of 70%, resulting in 152 valid cases. Data exploration involved a descriptive analysis, followed by a reliability estimation and an exploratory factorial analysis carried out through SPSS v.20. Confirmatory factorial analysis and path analysis (to examine relations between the different study factors) were studied using “R” software (Package 2.15.1., R Core Team, 2012) (Fox, 2006). Finally, in depth interviews were carried out to several marketing and sales managers in each of the six countries so as to further confirm the hypothesis and their relations. The models results prove that communication frequency has a positive impact on the quality of the same, which in turn has direct effects on the marketing-sales relations. Additionally, communication quality has an impact on the cognitive trust, which also relates not only to affective trust, but also to the relation between both areas. This means that in order to improve the implementation of innovation, managers should strive to enforce marketing-sales relations, facilitating the interpersonal trust construction (firstly cognitive, followed by affective trust), increasing the communication frequency, and therefore nurturing the communication quality among both areas. Through the second model, the results confirm the importance of creating effective relationships between sales and marketing to facilitate planning integrated new product implementations. While formalized new product development processes provide opportunities for sales and marketing to communicate, this does not directly influence the planning of integrated new product implementations. By using these formal opportunities to communicate to create information quality, it is possible to improve sales and marketing’s ability to integrate information during the planning process. Further, communication quality creates inter-personal trust in the other party’s competences (cognitive-based trust), leading to affect-based trust. Affect-based inter-personal trust, not only to improve the overall effectiveness of the sales and marketing relationship, but also helps in planning integrated new product implementations. This study contributes to the understanding of factors which enterprises can use to improve the inter-functional relations between marketing and sales, and the implementation of innovation in FMCG companies. The contribution of this investigation can be measured in two ways: enrichment of management and contribution to the academic area. From a business perspective, it provides massive consumption businesses leaders with knowledge on which factors affect innovation implementation, which results on mid and long-term success for the company. From an academic point of view, it provides knowledge on a prototype of successful innovation implementation management based on the marketing-sales interface effectiveness through a case study in the FMCG consumption market. Last but not least, it incorporates for the first time an empiric study on emerging geographies capable of recovery post a deep economic crisis through successful innovation implementation on their markets.

O EFEITO DO CONTROLE RESPIRATÓRIO EM VARIÁVEIS ELETROFISIOLÓGICAS DA ATENÇÃO

A prática do ioga tem se tornado cada vez mais popular, não apenas pelos benefícios físicos, mas principalmente pelo bem-estar psicológico trazido pela sua prática. Um dos componentes do ioga é o Prãnãyama, ou controle da respiração. A atenção e a respiração são dois mecanismos fisiológicos e involuntários requeridos para a execução do Prãnãyama. O principal objetivo desse estudo foi verificar se variáveis contínuas do EEG (potência de diferentes faixas que o compõem) seriam moduladas pelo controle respiratório, comparando-se separadamente as duas fases do ciclo respiratório (inspiração e expiração), na situação de respiração espontânea e controlada. Fizeram parte do estudo 19 sujeitos (7 homens/12 mulheres, idade média de 36,89 e DP = ± 14,46) que foram convidados a participar da pesquisa nas dependências da Faculdade de Saúde da Universidade Metodista de São Paulo. Para o registro do eletroencefalograma foi utilizado um sistema de posicionamento de cinco eletrodos Ag AgCl (FPz, Fz, Cz, Pz e Oz) fixados a uma touca de posicionamento rápido (Quick-Cap, Neuromedical Supplies®), em sistema 10-20. Foram obtidos valores de máxima amplitude de potência (espectro de potência no domínio da frequência) nas frequências teta, alfa e beta e delta e calculada a razão teta/beta nas diferentes fases do ciclo respiratório (inspiração e expiração), separadamente, nas condições de respiração espontânea e de controle respiratório. Para o registro do ciclo respiratório, foi utilizada uma cinta de esforço respiratório M01 (Pletismógrafo). Os resultados mostram diferenças significativas entre as condições de respiração espontânea e de controle com valores das médias da razão teta/beta menores na respiração controlada do que na respiração espontânea e valores de média da potência alfa sempre maiores no controle respiratório. Diferenças significativas foram encontradas na comparação entre inspiração e expiração da respiração controlada com diminuição dos valores das médias da razão teta/beta na inspiração e aumento nos valores das médias da potência alfa, sobretudo na expiração. Os achados deste estudo trazem evidências de que o controle respiratório modula variáveis eletrofisiológicas relativas à atenção refletindo um estado de alerta, porém mais relaxado do que na situação de respiração espontânea.

Potassium homeostasis in vacuolate plant cells.

Plant cells contain two major pools of K+, one in the vacuole and one in the cytosol. The behavior of K+ concentrations in these pools is fundamental to understanding the way this nutrient affects plant growth. Triple-barreled microelectrodes have been used to obtain the first fully quantitative measurements of the changes in K+ activity (aK) in the vacuole and cytosol of barley (Hordeum vulgare L.) root cells grown in different K+ concentrations. The electrodes incorporate a pH-selective barrel allowing each measurement to be assigned to either the cytosol or vacuole. The measurements revealed that vacuolar aK declined linearly with decreases in tissue K+ concentration, whereas cytosolic aK initially remained constant in both epidermal and cortical cells but then declined at different rates in each cell type. An unexpected finding was that cytoplasmic pH declined in parallel with cytosolic aK, but acidification of the cytosol with butyrate did not reveal any short-term link between these two parameters. These measurements show the very different responses of the vacuolar and cytosolic K+ pools to changes in K+ availability and also show that cytosolic K+ homeostasis differs quantitatively in different cell types. The data have been used in thermodynamic calculations to predict the need for, and likely mechanisms of, active K+ transport into the vacuole and cytosol. The direction of active K+ transport at the vacuolar membrane changes with tissue K+ status.

Human immunodeficiency virus (HIV) type 2-mediated inhibition of HIV type 1: a new approach to gene therapy of HIV-infection.

Human immunodeficiency virus (HIV) type 2, the second AIDS-associated human retrovirus, differs from HIV-1 in its natural history, infectivity, and pathogenicity, as well as in details of its genomic structure and molecular behavior. We report here that HIV-2 inhibits the replication of HIV-1 at the molecular level. This inhibition was selective, dose-dependent, and nonreciprocal. The closely related simian immunodeficiency provirus also inhibited HIV-1. The selectivity of inhibition was shown by the observation that HIV-2 did not significantly downmodulate the expression of the unrelated murine leukemia virus; neither did the murine leukemia virus markedly affect HIV-1 or HIV-2 expression. Moreover, while HIV-2 potently inhibited HIV-1, the reverse did not happen, thus identifying yet another and remarkable difference between HIV-1 and HIV-2. Mutational analysis of the HIV-2 genome suggested that the inhibition follows a complex pathway, possibly involving multiple genes and redundant mechanisms. Introduction of inactivating mutations into the structural and regulatory/accessory genes did not render the HIV-2 provirus ineffective. Some of the HIV-2 gene defects, such as that of tat and rev genes, were phenotypically transcomplemented by HIV-1. The HIV-2 proviruses with deletions in the putative packaging signal and defective for virus replication were effective in inducing the suppressive phenotype. Though the exact mechanism remains to be defined, the inhibition appeared to be mainly due to an intracellular molecular event because it could not be explained solely on the basis of cell surface receptor mediated interference. The results support the notion that the inhibition likely occurred at the level of viral RNA, possibly involving competition between viral RNAs for some transcriptional factor essential for virus replication. Induction of a cytokine is another possibility. These findings might be relevant to the clinical-epidemiological data suggesting that infection with HIV-2 may offer some protection against HIV-1 infection.

Contribuição das células-tronco mesenquimais para as propriedades tumorigênicas de células de glioblastoma humano

Células-tronco mesenquimais (CTM) apresentam tropismo a tumores, sendo importantes componentes do estroma tumoral. No cérebro, o nicho perivascular é uma importante fonte de CTM, as quais podem contribuir direta e/ou indiretamente para o desenvolvimento de tumores, embora os mecanismos envolvidos sejam pouco conhecidos. No presente trabalho, investigou-se a influência de CTM sobre a proliferação, capacidade invasiva e tumorigenicidade de células de Glioblastoma (GBM) humano. Sabe-se que CTM produzem TGFB1, uma citocina multifuncional envolvida em imunomodulação, proliferação, migração e transição epitelial-mesenquimal de células tumorais. Experimentos in vitro, realizados com meios condicionados de CTM de cordão umbilical humano com silenciamento permanente do gene TGFB1, demonstraram que o TGFB1 secretado por CTM é capaz de aumentar significativamente a proliferação e viabilidade de células de GBM humano da linhagem U87FP635. Esses resultados revelam uma importante ação parácrina dessa citocina regulatória, quando produzida por outros tipos celulares contidos no microambiente tumoral. Entretanto, sob condições experimentais que melhor mimetizam o microambiente tumoral, detectou-se que CTM também afetam o comportamento de células tumorais por um mecanismo alternativo, dependente de contato celular, mas independente dos níveis de TGFB1 secretados pelas CTM. Sob condições de cocultivo celular, envolvendo contato físico entre CTM e células de GBM U87FP635, detectou-se um aumento significativo na quantidade de células tumorais viáveis. Quando cultivadas na forma de esferoides tumorais, o contato com CTM aumentou a capacidade invasiva das células U87FP635. Finalmente, em modelo in vivo ectópico de GBM, células U87FP635 geraram tumores mais desenvolvidos quando coinjetadas com CTM. Esses efeitos pró-tumorigênicos foram observados tanto em contato com CTM controles, quanto com CTM contendo o gene TGFB1 permanentemente silenciado. Assim, esses achados indicam que CTM podem exercer efeitos pró-tumorigênicos por dois mecanismos alternativos e independentes: ação parácrina de TGFB1 secretado por CTM e ação mediada por contato célula-célula. Nas condições experimentais testadas, o mecanismo dependente de contato célula-célula demonstrou ser predominante. O estudo proteômico do secretoma dessas células identificou 126 proteínas diferencialmente expressas além de 10 proteínas exclusivamente detectadas em meios condicionados de cocultivos de CTM com células de GBM U87FP635. Cerca de 80% dessas proteínas exclusivamente secretadas pelo contato célula-célula são componentes de exossomos e estão envolvidas em proliferação celular e desenvolvimento tecidual. Esses resultados apontam uma interação dinâmica de comunicação entre CTM e células tumorais, e revelam algumas proteínas interessantes potencialmente envolvidas em uma ação pró-tumorigênica de CTM mediada por contato celular

Comportamento tribo-mecânico e desgaste adesivo de materiais em nanoescala: análises por dinâmica molecular e mecânica do contínuo.

As formulações baseadas na mecânica do contínuo, embora precisas até certo ponto, por vezes não podem ser utilizadas, ou não são conceitualmente corretas para o entendimento de fenômenos em escalas reduzidas. Estas limitações podem aparecer no estudo dos fenômenos tribológicos em escala nanométrica, que passam a necessitar de novos métodos experimentais, teóricos e computacionais que permitam explorar estes fenômenos com a resolução necessária. Simulações atomísticas são capazes de descrever fenômenos em pequena escala, porém, o número necessário de átomos modelados e, portanto, o custo computacional - geralmente torna-se bastante elevado. Por outro lado, os métodos de simulação associados à mecânica do contínuo são mais interessantes em relação ao custo computacional, mas não são precisos na escala atômica. A combinação entre essas duas abordagens pode, então, permitir uma compreensão mais realista dos fenômenos da tribologia. Neste trabalho, discutem-se os conceitos básicos e modelos de atrito em escala atômica e apresentam-se estudos, por meio de simulação numérica, para a análise e compreensão dos mecanismos de atrito e desgaste no contato entre materiais. O problema é abordado em diferentes escalas, e propõe-se uma abordagem conjunta entre a Mecânica do Contínuo e a Dinâmica Molecular. Para tanto, foram executadas simulações numéricas, com complexidade crescente, do contato entre superfícies, partindo-se de um primeiro modelo que simula o efeito de defeitos cristalinos no fenômeno de escorregamento puro, considerando a Dinâmica Molecular. Posteriormente, inseriu-se, nos modelos da mecânica do contínuo, considerações sobre o fenômeno de adesão. A validação dos resultados é feita pela comparação entre as duas abordagens e com a literatura.

Comportamento anti-social e delinquência na adolescência

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Mechanisms of Mycoplasma-induced inflammation and cellular transformation

There is a growing interest in “medical gasses” for their antibacterial and anti-inflammatory properties. Hydrogen sulfide (H2S), a member of the family of gasotransmitters, is in fact increasingly being recognized as an important signaling molecule, but its precise role in the regulation of the inflammatory response is still not clear. For this reason, the aim of the first part of this thesis was to investigate the effects of H2S on the expression of pro-inflammatory cytokines, such as MCP-1, by using an in vitro model composed by both primary monocytes-derived macrophages cultures and the human monocytic cell line U937 infected with Mycoplasma fermentans, a well-known pro-inflammatory agent. In our experiments, we observed a marked increase in the production of pro-inflammatory cytokines in infected cells. In particular, MCP-1 was induced both at the RNA and at the protein level. To test the effects of H2S on infected cells, we treated the cells with two different H2S donors (NaHS and GYY4137), showing that both H2S treatments had anti-inflammatory effects in Mycoplasma-infected cells: the levels of MCP-1, both mRNA expression and protein production, were reduced. Our subsequent studies aimed at understanding the molecular mechanisms responsible for these effects, focused on two specific molecular pathways, both involved in inflammation: the NF-κB and the Nrf2 pathway. After treatment with pharmacological inhibitors, we demonstrated that Mycoplasma fermentans induces MCP-1 expression through the TLR-NF-κB pathway with the nuclear translocation of its subunits, while treatment with H2S completely blocked the nuclear translocation of NF-κB heterodimer p65/p50. Then, once infected cells were treated with H2S donors, we observed an increased protective effect of Nrf2 and also a decrease in ROS production. These results highlight the importance of H2S in reducing the inflammatory process caused by Mycoplasma fermentans. To this regard, it should be noted that several projects are currently ongoing to develop H2S-releasing compounds as candidate drugs capable of alleviating cell deterioration and to reduce the rate of decline in organ function. In the second part of this study, we investigated the role of Mycoplasma infection in cellular transformation. Infectious agents are involved in the etiology of many different cancers and a number of studies are still investigating the role of microbiota in tumor development. Mycoplasma has been associated with some human cancers, such as prostate cancer and non-Hodgkin’s lymphoma in HIV-seropositive people, and its potential causative role and molecular mechanisms involved are being actively investigated. To this regard, in vitro studies demonstrated that, upon infection, Mycoplasma suppresses the transcriptional activity of p53, key protein in the cancer suppression. As a consequence, infected cells were less susceptible to apoptosis and proliferated more than the uninfected cells. The mechanism(s) responsible for the Mycoplasma-induced inhibitory effect on p53 were not determined. Aim of the second part of this thesis was to better understand the tumorigenic role of the microorganism, by investigating more in details the effect(s) of Mycoplasma on p53 activity in an adenocarcinoma HCT116 cell line. Treatment of Mycoplasma-infected cells with 5FU or with Nutlin, two molecules that induce p53 activity, resulted in cellular proliferation comparable to untreated controls. These results suggested that Mycoplasma infection inhibited p53 activity. Immunoprecipitation of p53 with specific antibodies, and subsequent Gas Chromatography and Mass Spectroscopy (GC-MS) assays, allowed us to identify several Mycoplasma-specific proteins interacting with p53, such as DnaK, a prokaryotic heat shock protein and stress inducible chaperones. In cells transfected with DnaK we observed i) reduced p53 protein levels; ii) reduced activity and expression of p21, Bax and PUMA, iii) a marked increase in cells leaving G1 phase. Taken together, these data show an interaction between the human p53 and the Mycoplasma protein DnaK, with the consequent decreased p53 activity and decreased capability to respond to DNA damage and prevent cell proliferation. Our data indicate that Mycoplasma could be involved in cancer formation and the mechanism(s) has the potential to be a target for cancer diagnosis and treatment(s).

O EFEITO DO CONTROLE RESPIRATÓRIO EM VARIÁVEIS ELETROFISIOLÓGICAS DA ATENÇÃO

A prática do ioga tem se tornado cada vez mais popular, não apenas pelos benefícios físicos, mas principalmente pelo bem-estar psicológico trazido pela sua prática. Um dos componentes do ioga é o Prãnãyama, ou controle da respiração. A atenção e a respiração são dois mecanismos fisiológicos e involuntários requeridos para a execução do Prãnãyama. O principal objetivo desse estudo foi verificar se variáveis contínuas do EEG (potência de diferentes faixas que o compõem) seriam moduladas pelo controle respiratório, comparando-se separadamente as duas fases do ciclo respiratório (inspiração e expiração), na situação de respiração espontânea e controlada. Fizeram parte do estudo 19 sujeitos (7 homens/12 mulheres, idade média de 36,89 e DP = ± 14,46) que foram convidados a participar da pesquisa nas dependências da Faculdade de Saúde da Universidade Metodista de São Paulo. Para o registro do eletroencefalograma foi utilizado um sistema de posicionamento de cinco eletrodos Ag AgCl (FPz, Fz, Cz, Pz e Oz) fixados a uma touca de posicionamento rápido (Quick-Cap, Neuromedical Supplies®), em sistema 10-20. Foram obtidos valores de máxima amplitude de potência (espectro de potência no domínio da frequência) nas frequências teta, alfa e beta e delta e calculada a razão teta/beta nas diferentes fases do ciclo respiratório (inspiração e expiração), separadamente, nas condições de respiração espontânea e de controle respiratório. Para o registro do ciclo respiratório, foi utilizada uma cinta de esforço respiratório M01 (Pletismógrafo). Os resultados mostram diferenças significativas entre as condições de respiração espontânea e de controle com valores das médias da razão teta/beta menores na respiração controlada do que na respiração espontânea e valores de média da potência alfa sempre maiores no controle respiratório. Diferenças significativas foram encontradas na comparação entre inspiração e expiração da respiração controlada com diminuição dos valores das médias da razão teta/beta na inspiração e aumento nos valores das médias da potência alfa, sobretudo na expiração. Os achados deste estudo trazem evidências de que o controle respiratório modula variáveis eletrofisiológicas relativas à atenção refletindo um estado de alerta, porém mais relaxado do que na situação de respiração espontânea.

Ocular biotribology and contact lens lubrication mechanisms

The work described in this thesis is concerned with mechanisms of contact lens lubrication. There are three major driving forces in contact lens design and development; cost, convenience, and comfort. Lubrication, as reflected in the coefficient of friction, is becoming recognised as one of the major factors affecting the comfort of the current generation of contact lenses, which have benefited from several decades of design and production improvements. This work started with the study of the in-eye release of soluble macromolecules from a contact lens matrix. The vehicle for the study was the family of CIBA Vision Focus® DAILIES® daily disposable contact lenses which is based on polyvinyl alcohol (PVA). The effective release of linear soluble PVA from DAILIES on the surface of the lens was shown to be beneficial in terms of patient comfort. There was a need to develop a novel characterisation technique in order to study these effects at surfaces; this led to the study of a novel tribological technique, which allowed the friction coefficients of different types of contact lenses to be measured reproducibly at genuinely low values. The tribometer needed the ability to accommodate the following features: (a) an approximation to eye lid load, (b) both new and ex-vivo lenses, (c) variations in substrate, (d) different ocular lubricants (including tears). The tribometer and measuring technique developed in this way was used to examine the surface friction and lubrication mechanisms of two different types of contact lenses: daily disposables and silicone hydrogels. The results from the tribometer in terms of both mean friction coefficient and the friction profiles obtained allowed various mechanisms used for surface enhancement now seen in the daily disposable contact lens sector to be evaluated. The three major methods used are: release of soluble macromolecules (such as PVA) from the lens matrix, irreversible surface binding of a macromolecule (such as polyvinyl pyrrolidone) by charge transfer and the simple polymer adsorption (e.g. Pluoronic) at the lens surface. The tribological technique was also used to examine the trends in the development of silicone hydrogel contact lenses. The focus of the principles in the design of silicone hydrogels has now shifted from oxygen permeability, to the improvement of surface properties. Presently, tribological studies reflect the most effective in vitro method of surface evaluation in relation to the in-eye comfort.

Characterization of oxide layers grown on D9 austenitic stainless steel in lead bismuth eutectic

Lead bismuth eutectic (LBE) is a possible coolant for fast reactors and targets in spallation neutron sources. Its low melting point, high evaporation point, good thermal conductivity, low reactivity, and good neutron yield make it a safe and high performance coolant in radiation environments. The disadvantage is that it is a corrosive medium for most steels and container materials. This study was performed to evaluate the corrosion behavior of the austenitic stainless steel D9 in oxygen controlled LBE. In order to predict the corrosion behavior of steel in this environment detailed analyses have to be performed on the oxide layers formed on these materials and various other relevant materials upon exposure to LBE. In this study the corrosion/oxidation of D9 stainless steel in LBE was investigated in great detail. The oxide layers formed were characterized using atomic force microscopy, magnetic force microscopy, nanoindentation, and scanning electron microscopy with wavelength-dispersive spectroscopy (WDS) to understand the corrosion and oxidation mechanisms of D9 stainless steel in contact with the LBE. What was previously believed to be a simple double oxide layer was identified here to consist of at least 4 different oxide layers. It was found that the inner most oxide layer takes over the grain structure of what used to be the bulk steel material while the outer oxide layer consists of freshly grown oxides with a columnar structure. These results lead to a descriptive model of how these oxide layers grow on this steel under the harsh environments encountered in these applications.

KM governance:the mechanisms for guiding and controlling KM program

Purpose: To establish and sustain their KM programs organizations need to establish mechanisms to ensure their governance. KM programs require business integration, senior management involvement and decision making authority. The present research investigates the KM governance mechanisms organizations use to guide and control their KM programs. The research seeks to contribute to a better understanding of the governance of KM and to support organizations in the development of their KM programs. Methodology: The study employs multiple case research methodology to analyze the KM governance arrangements of twelve international organizations and identify patterns in their governance configurations. Findings: The analysis identifies a range of structural, process and relational mechanisms that are critical for governing an organizational KM program. Different patterns among the KM governance mechanisms are identified which lead to the development of generic KM governance typologies. Research implications: The development of the KM governance framework allows future research to systematically investigate the KM governance phenomenon. As the present study is based on a configurational analysis future research should particularly target the performance implications of different KM governance configurations. Practical implications: The research provides insights into the diversity of KM governance mechanisms and their impact on a KM program. The KM governance framework can assist managers in reviewing their present and prospective KM programs and thereby support benchmarking or re-organization efforts. Originality: Building on prior research that has focused on individual KM governance aspects, the present study adopts a comprehensive perspective integrating structural, process and relational governance mechanisms.

Breaking promises. The Hungarian experience

My main aim is to present the phenomena related to fulfilment and breach of promises and the economic, political and ethical problems arising from these. I discuss questions that we all meet with in daily life and see mentioned in the press, other forums of public discourse, gatherings of friends, or sessions of Parliament. There are some who complain that a building contractor has not done a renovation job properly according to contract. Economists argue over the outcome of late repayments on loans advanced for purchasing real estate. Opposition meetings chide the governing party over unfulfilled campaign promises. I am seeking what is common among these seemingly different cases. Can we see identical or similar behavior patterns and social mechanisms in them? Do they lead to similar decision-making dilemmas and reactions?