Biological Activity of Rhodium Metalloinsertors and the Design of Bifunctional Conjugates

The Barton laboratory has established that octahedral rhodium complexes bearing the sterically expansive 5,6-chrysene diimine ligand can target thermodynamically destabilized sites, such as base pair mismatches, in DNA with high affinity and selectivity. These complexes approach DNA from the minor groove, ejecting the mismatched base pairs from the duplex in a binding mode termed metalloinsertion. In recent years, we have shown that these metalloinsertor complexes also exhibit cytotoxicity preferentially in cancer cells that are deficient in the mismatch repair (MMR) machinery.

Here, we establish that a sensitive structure-activity relationship exists for rhodium metalloinsertors. We studied the relationship between the chemical structures of metalloinsertors and their effect on biological activity for ten complexes with similar DNA binding affinities, but wide variation in their lipophilicity. Drastic differences were observed in the selectivities of the complexes for MMR-deficient cells. Compounds with hydrophilic ligands were highly selective, exhibiting preferential cytotoxicity in MMR-deficient cells at low concentrations and short incubation periods, whereas complexes with lipophilic ligands displayed poor cell-selectivity. It was discovered that all of the complexes localized to the nucleus in concentrations sufficient for mismatch binding; however, highly lipophilic complexes also exhibited high mitochondrial uptake. Significantly, these results support the notion that mitochondrial DNA is not the desired target for our metalloinsertor complexes; instead, selectivity stems from targeting mismatches in genomic DNA.

We have also explored the potential for metalloinsertors to be developed into more complex structures with multiple functionalities that could either enhance their overall potency or impart mismatch selectivity onto other therapeutic cargo. We have constructed a family of bifunctional metalloinsertor conjugates incorporating cis-platinum, each unique in its chemical structure, DNA binding interactions, and biological activity. The study of these complexes in MMR-deficient cells has established that the cell-selective biological activity of rhodium metalloinsertors proceeds through a critical cellular pathway leading to necrosis.

We further explored the underlying mechanisms surrounding the biological response to mismatch recognition by metalloinsertors in the genome. Immunofluorescence assays of MMR-deficient and MMR-proficient cells revealed that a critical biomarker for DNA damage, phosphorylation of histone H2AX (γH2AX) rapidly accumulates in response to metalloinsertor treatment, signifying the induction of double strand breaks in the genome. Significantly, we have discovered that our metalloinsertor complexes selectively inhibit transcription in MMR-deficient cells, which may be a crucial checkpoint in the eventual breakdown of the cell via necrosis. Additionally, preliminary in vivo studies have revealed the capability of these compounds to traverse the complex environments of multicellular organisms and accumulate in MMR-deficient tumors. Our ever-increasing understanding of metalloinsertors, as well as the development of new generations of complexes both monofunctional and bifunctional, enables their continued progress into the clinic as promising new chemotherapeutic agents.

Análise dinâmica de plataformas de aço para produção de petróleo com base na consideração do efeito da interação solo-estrutura

Atualmente, as tendências competitivas do mercado mundial, têm forçado os engenheiros estruturais a desenvolver soluções de projeto que acarretem em menor peso e custo de execução. Uma consequência direta desta nova tendência de projeto é o aumento considerável de problemas relacionados a vibrações de piso indesejadas. Por esta razão, os sistemas estruturais de pisos podem tornar-se vulneráveis a vibrações excessivas, como por exemplo, aquelas induzidas por equipamentos mecânicos (máquinas rotativas). Deste modo, este trabalho objetiva investigar o comportamento dinâmico de uma plataforma de aço para produção de petróleo, localizada na bacia de Santos (campo de Merluza), São Paulo, Brasil. Para tal, investiga-se a influência das ações dinâmicas oriundas dos equipamentos mecânicos localizados sobre os decks metálicos da plataforma. A resposta dinâmica do modelo estrutural foi determinada através de um extenso estudo numérico, a partir da análise de suas frequências naturais, deslocamentos, velocidades e acelerações de pico. Nesta investigação, as cargas dinâmicas provenientes dos equipamentos mecânicos (máquinas rotativas) foram aplicadas sobre o piso metálico do sistema estrutural. Com base obtenção da resposta dinâmica da estrutura (deslocamentos, velocidades e acelerações), foi possível avaliar a performance do modelo estrutural em termos de critérios de conforto humano e das tolerâncias máximas referentes aos equipamentos mecânicos, de acordo com normas e recomendações de projeto.

Informe técnico sobre posibles alternativas de valorización energética de residuos forestales

[ES]Mediante este proyecto, se trata de buscar diferentes alternativas de valorización energética de residuos forestales, analizar su proceso y los equipos involucrados, y al final elegir la más adecuada en base a ciertos criterios.

Influencia de la internacionalización en el desempeño social de las empresas

[ES] Recientemente estudios previos han comenzado a plantear la posible relación entre el fenómeno de la internacionalización y responsabilidad social corporativa de las organizaciones. En este trabajo analizamos en qué medida la internacionalización contribuye a una mejora sustancial del desempeño social de las empresas. Usando una muestra de 102 empresas estadounidenses de los sectores químico, energético y de maquinaria industrial, nuestros resultados muestran que no todas las dimensiones de la internacionalización permiten a las empresas adquirir conocimiento valioso que conduzca a una mejora de su desempeño social. Concretamente, comprobamos que, mientras el porcentaje de ventas en mercados extranjeros y la diversificación internacional cultural favorecen la puesta en marcha de prácticas de responsabilidad social corporativa avanzadas que redundan en un mejor desempeño social (cumpliendo con las demandas sociales de los grupos de interés de los distintos mercados donde operan), la diversificación internacional geográfica no tiene efecto directo en el desempeño social de las empresas. Por tanto, no es tan importante el número de mercados donde la empresa actúa, sino el grado de diferenciación cultural existente entre los mismos lo que hace que la propia empresa mejore su desempeño social. El presente trabajo incluye implicaciones relevantes para el mundo académico, directivos y reguladores públicos.

Leukemia-Associated Mutations in Nucleophosmin Alter Recognition by CRM1: Molecular Basis of Aberrant Transport

Nucleophosmin (NPM) is a nucleocytoplasmic shuttling protein, normally enriched in nucleoli, that performs several activities related to cell growth. NPM mutations are characteristic of a subtype of acute myeloid leukemia (AML), where mutant NPM seems to play an oncogenic role. AML-associated NPM mutants exhibit altered subcellular traffic, being aberrantly located in the cytoplasm of leukoblasts. Exacerbated export of AML variants of NPM is mediated by the nuclear export receptor CRM1, and due, in part, to a mutationally acquired novel nuclear export signal (NES). To gain insight on the molecular basis of NPM transport in physiological and pathological conditions, we have evaluated the export efficiency of NPM in cells, and present new data indicating that, in normal conditions, wild type NPM is weakly exported by CRM1. On the other hand, we have found that AML-associated NPM mutants efficiently form complexes with CRM1HA (a mutant CRM1 with higher affinity for NESs), and we have quantitatively analyzed CRM1HA interaction with the NES motifs of these mutants, using fluorescence anisotropy and isothermal titration calorimetry. We have observed that the affinity of CRM1HA for these NESs is similar, which may help to explain the transport properties of the mutants. We also describe NPM recognition by the import machinery. Our combined cellular and biophysical studies shed further light on the determinants of NPM traffic, and how it is dramatically altered by AML-related mutations.

Latt-Mayer, um estudo de caso: tecnologia na história do design gráfico brasileiro.

Nosso objetivo neste trabalho é investigar como o design gráfico se relaciona com os meios de produção da indústria gráfica brasileira. Não é comum encontrar referências abordando temas que relacionam design e tecnologia em livros sobre a história do Brasil, o que demonstra a urgência do resgate histórico de indústrias que implementaram novas tecnologias para atender a demandas projetuais e mercadológicas. Sediada na cidade do Rio de Janeiro, a clicheria Latt-Mayer como era comumente chamada, era sinônimo de qualidade e tecnologia. Entre seus clientes podemos apontar agências de publicidade, editoras, escritórios de designers e gráficas de todo país. Esta pesquisa se baseia em um corpus composto por imagens, documentos, manuais técnicos, reportagens, matrizes e depoimentos com pessoas que estiveram diretamente envolvidas com essa empresa. Catalogamos maquinaria e técnicas utilizadas, além de projetos gráficos relevantes que tiveram matrizes e provas produzidas na empresa. Por meio deste trabalho, é possível visualizar um abrangente painel histórico, que contempla um período em que ocorreram significativas transformações na relação entre designers e tecnologia na indústria gráfica brasileira.

Diseño de una tarjeta de drivers para un inversor de potencia.

[EN]The purpose of this project is to design a system which improves the e ciency of power inverters. The project is focused in the analysis of the di erent power semiconductors (based on silicon, silicon carbide and gallium nitride) and driver applications. This system can be implemented in di erent future versions of power inverters and in many kind of applications like electrical vehicles. Other than that, it can be also implemented on any machinery requiring an inverter obtaining more energy and reduce manufacturing costs.

Visibility recovery on images acquired in attenuating media. Application to underwater, fog, and mammographic imaging

136 p.

Development of nucleic acid vaccines: use of self-amplifying RNA in lipid nanoparticles

Self-amplifying RNA or RNA replicon is a form of nucleic acid-based vaccine derived from either positive-strand or negative-strand RNA viruses. The gene sequences encoding structural proteins in these RNA viruses are replaced by mRNA encoding antigens of interest as well as by RNA polymerase for replication and transcription. This kind of vaccine has been successfully assayed with many different antigens as vaccines candidates, and has been shown to be potent in several animal species, including mice, nonhuman primates, and humans. A key challenge to realizing the broad potential of self-amplifying vaccines is the need for safe and effective delivery methods. Ideally, an RNA nanocarrier should provide protection from blood nucleases and extended blood circulation, which ultimately would increase the possibility of reaching the target tissue. The delivery system must then be internalized by the target cell and, upon receptor-mediated endocytosis, must be able to escape from the endosomal compartment into the cell cytoplasm, where the RNA machinery is located, while avoiding degradation by lysosomal enzymes. Further, delivery systems for systemic administration ought to be well tolerated upon administration. They should be safe, enabling the multiadministration treatment modalities required for improved clinical outcomes and, from a developmental point of view, production of large batches with reproducible specifications is also desirable. In this review, the concept of self-amplifying RNA vaccines and the most promising lipid-based delivery systems are discussed.

Adenylate Cyclase Toxin promotes bacterial internalisation into non phagocytic cells

Bordetella pertussis causes whooping cough, a respiratory infectious disease that is the fifth largest cause of vaccine-preventable death in infants. Though historically considered an extracellular pathogen, this bacterium has been detected both in vitro and in vivo inside phagocytic and non-phagocytic cells. However the precise mechanism used by B. pertussis for cell entry, or the putative bacterial factors involved, are not fully elucidated. Here we find that adenylate cyclase toxin (ACT), one of the important toxins of B. pertussis, is sufficient to promote bacterial internalisation into non-phagocytic cells. After characterization of the entry route we show that uptake of "toxin-coated bacteria" proceeds via a clathrin-independent, caveolae-dependent entry pathway, allowing the internalised bacteria to survive within the cells. Intracellular bacteria were found inside non-acidic endosomes with high sphingomyelin and cholesterol content, or "free" in the cytosol of the invaded cells, suggesting that the ACT-induced bacterial uptake may not proceed through formation of late endolysosomes. Activation of Tyr kinases and toxin-induced Ca2+-influx are essential for the entry process. We hypothesize that B. pertussis might use ACT to activate the endocytic machinery of non-phagocytic cells and gain entry into these cells, in this way evading the host immune system.

A catch-free stock assessment model with application to goliath grouper (Epinephelus itajara) off southern Florida

Many modern stock assessment methods provide the machinery for determining the status of a stock in relation to certain reference points and for estimating how quickly a stock can be rebuilt. However, these methods typically require catch data, which are not always available. We introduce a model-based framework for estimating reference points, stock status, and recovery times in situations where catch data and other measures of absolute abundance are unavailable. The specif ic estimator developed is essentially an age-structured production model recast in terms relative to pre-exploitation levels. A Bayesian estimation scheme is adopted to allow the incorporation of pertinent auxiliary information such as might be obtained from meta-analyses of similar stocks or anecdotal observations. The approach is applied to the population of goliath grouper (Epinephelus itajara) off southern Florida, for which there are three indices of relative abundance but no reliable catch data. The results confirm anecdotal accounts of a marked decline in abundance during the 1980s followed by a substantial increase after the harvest of goliath grouper was banned in 1990. The ban appears to have reduced fishing pressure to between 10% and 50% of the levels observed during the 1980s. Nevertheless, the predicted fishing mortality rate under the ban appears to remain substantial, perhaps owing to illegal harvest and depth-related release mortality. As a result, the base model predicts that there is less than a 40% chance that the spawning biomass will recover to a level that would produce a 50% spawning potential ratio.