A Burkholderia cenocepacia gene encoding a non-functional tyrosine phosphatase is required for the delayed maturation of the bacteria-containing vacuoles in macrophages

Burkholderia cenocepacia infects patients with cystic fibrosis. We have previously shown that B. cenocepacia can survive in macrophages within membrane vacuoles (BcCVs) that preclude fusion with the lysosome. The bacterial factors involved in B. cenocepacia intracellular survival are not fully elucidated. We report here that deletion of BCAM0628, encoding a predicted low-molecular weight protein tyrosine phosphatase (LMW-PTP) that is restricted to B. cenocepacia strains of the transmissible ET-12 clone, accelerates the maturation of the BcCVs. Compared to parental strain and deletion mutants in other LMW-PTPs that are widely conserved in Burkholderia species, a greater proportion of BcCVs containing the BCAM0628 mutant were targeted to the lysosome. Accelerated BcCV maturation was not due to reduced intracellular viability since BCAM0628 survived and replicated in macrophages similarly to the parental strain. Therefore, BCAM0628 was referred to as dpm (delayed phagosome maturation). We provide evidence that the Dpm protein is secreted during growth in vitro and upon macrophage infection. Dpm secretion requires an N-terminal signal peptide. Heterologous expression of Dpm in B. multivorans confers to this bacterium a similar phagosomal maturation delay as found with B. cenocepacia. We demonstrate that Dpm is an inactive phosphatase, suggesting that its contribution to phagosomal maturation arrest must be unrelated to tyrosine phosphatase activity.

Test methods for concrete durability indicators

Durability of concrete structures is primarily dependent on the environmental influences, i.e. the penetration of aggressive substances in the structural element from the environment. Penetrability is an important durability indicator of concrete and by specifying different classes of penetrability of concrete it should be possible to design a structure with the required resistance to environmental loads. This chapter covers descriptions of the available and commonly applied in situ and laboratory, non-invasive and semi-invasive test methods for evaluating concrete penetrability properties.

Application examples of performance-based specification and quality control

This chapter presents an overview on several performance-based approaches for concrete durability specification and conformity assessment of the as-built structure.

Enhancing design space exploration by extending CPU/GPU specifications onto FPGAs

The design cycle for complex special-purpose computing systems is extremely costly and time-consuming. It involves a multiparametric design space exploration for optimization, followed by design verification. Designers of special purpose VLSI implementations often need to explore parameters, such as optimal bitwidth and data representation, through time-consuming Monte Carlo simulations. A prominent example of this simulation-based exploration process is the design of decoders for error correcting systems, such as the Low-Density Parity-Check (LDPC) codes adopted by modern communication standards, which involves thousands of Monte Carlo runs for each design point. Currently, high-performance computing offers a wide set of acceleration options that range from multicore CPUs to Graphics Processing Units (GPUs) and Field Programmable Gate Arrays (FPGAs). The exploitation of diverse target architectures is typically associated with developing multiple code versions, often using distinct programming paradigms. In this context, we evaluate the concept of retargeting a single OpenCL program to multiple platforms, thereby significantly reducing design time. A single OpenCL-based parallel kernel is used without modifications or code tuning on multicore CPUs, GPUs, and FPGAs. We use SOpenCL (Silicon to OpenCL), a tool that automatically converts OpenCL kernels to RTL in order to introduce FPGAs as a potential platform to efficiently execute simulations coded in OpenCL. We use LDPC decoding simulations as a case study. Experimental results were obtained by testing a variety of regular and irregular LDPC codes that range from short/medium (e.g., 8,000 bit) to long length (e.g., 64,800 bit) DVB-S2 codes. We observe that, depending on the design parameters to be simulated, on the dimension and phase of the design, the GPU or FPGA may suit different purposes more conveniently, thus providing different acceleration factors over conventional multicore CPUs.

Burkholderia cenocepacia Lipopolysaccharide Modification and Flagellin Glycosylation Affect Virulence but Not Innate Immune Recognition in Plants

Burkholderia cenocepacia causes opportunistic infections in plants, insects, animals, and humans, suggesting that “virulence” depends on the host and its innate susceptibility to infection. We hypothesized that modifications in key bacterial molecules recognized by the innate immune system modulate host responses to B. cenocepacia. Indeed, modification of lipo- polysaccharide (LPS) with 4-amino-4-deoxy-L-arabinose and flagellin glycosylation attenuates B. cenocepacia infection in Arabi- dopsis thaliana and Galleria mellonella insect larvae. However, B. cenocepacia LPS and flagellin triggered rapid bursts of nitric oxide and reactive oxygen species in A. thaliana leading to activation of the PR-1 defense gene. These responses were drastically reduced in plants with fls2 (flagellin FLS2 host receptor kinase), Atnoa1 (nitric oxide-associated protein 1), and dnd1-1 (reduced production of nitric oxide) null mutations. Together, our results indicate that LPS modification and flagellin glycosylation do not affect recognition by plant receptors but are required for bacteria to establish overt infection.

ArnT proteins that catalyse the glycosylation of lipopolysaccharide share common features with bacterial N-oligosaccharyltransferases

ArnT is a glycosyltransferase that catalyses the addition of 4-amino-4-deoxy-L-arabinose (L-Ara4N) to the lipid A moiety of the lipopolysaccharide. This is a critical modification enabling bacteria to resist killing by antimicrobial peptides. ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. We also demonstrate that the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain is required for polymyxin B resistance in vivo. Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyse lipid A modification with L-Ara4N. An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence.

Tyrosine phosphorylation and dephosphorylation in Burkholderia cenocepacia affect biofilm formation, growth under nutritional deprivation, and pathogenicity

Burkholderia cenocepacia, a member of the B. cepacia complex (Bcc), is an opportunistic pathogen causing serious chronic infections in patients with cystic fibrosis. Tyrosine phosphorylation has emerged as an important post-translational modification modulating the physiology and pathogenicity of Bcc bacteria. Here, we investigated the predicted bacterial tyrosine kinases BCAM1331 and BceF, and the low molecular weight protein tyrosine phosphatases BCAM0208, BceD and BCAL2200 of B. cenocepacia K56-2. We show that BCAM1331, BceF, BCAM0208 and BceD contributed to biofilm formation, while BCAL2200 was required for growth in nutrient-limited conditions. Multiple deletions of either tyrosine kinase or low molecular weight protein tyrosine phosphatases genes resulted in attenuation of B. cenocepacia intramacrophage survival and reduced pathogenicity in the Galleria mellonella larvae infection model. Experimental evidence indicates that BCAM1331 displays a reduced
tyrosine autophosphorylation activity compared to BceF. Using the artificial substrate p-nitrophenyl phosphate, the phosphatase activity of the three low molecular weight protein tyrosine phosphatases demonstrated similar kinetic parameters. However, only BCAM0208 and BceD could dephosphorylate BceF. Further, BCAL2200 becomes tyrosine phosphorylated in vivo and catalyzes its auto-dephosphorylation. Together, our data suggest that despite having similar biochemical activities low molecular weight protein tyrosine phosphatases and tyrosine kinases have both overlapping and specific roles in the physiology of B. cenocepacia.

On the performance of LDPC and turbo decoder architectures with unreliable memories

In this paper, we investigate the impact of faulty memory bit-cells on the performance of LDPC and Turbo channel decoders based on realistic memory failure models. Our study investigates the inherent error resilience of such codes to potential memory faults affecting the decoding process. We develop two mitigation mechanisms that reduce the impact of memory faults rather than correcting every single error. We show how protection of only few bit-cells is sufficient to deal with high defect rates. In addition, we show how the use of repair-iterations specifically helps mitigating the impact of faults that occur inside the decoder itself.

Real-space grids and the Octopus code as tools for the development of new simulation approaches for electronic systems

Real-space grids are a powerful alternative for the simulation of electronic systems. One of the main advantages of the approach is the flexibility and simplicity of working directly in real space where the different fields are discretized on a grid, combined with competitive numerical performance and great potential for parallelization. These properties constitute a great advantage at the time of implementing and testing new physical models. Based on our experience with the Octopus code, in this article we discuss how the real-space approach has allowed for the recent development of new ideas for the simulation of electronic systems. Among these applications are approaches to calculate response properties, modeling of photoemission, optimal control of quantum systems, simulation of plasmonic systems, and the exact solution of the Schrödinger equation for low-dimensionality systems.

Geologia 11.º ano - guia do professor

Na primeira parte, são fundamentalmente as orientações metodologicas, as quais incluem referências à educação para o desenvolvimento sustentável, algumas sugestões didáticas e observações relativas à avalliação das aprendizagens. Pretende-se que os professores desenvolvam um quadro de referência, o qual será importante para justificar algumas das decisões que terão de tomar na preparação das suas aulas e, depois, em situação de ato de ensino. Na segunda parte, são apresentados para cada uma das quatro Unidades Temáticas que integram o programa de Geologia do 11.º ano, mapas de conceitos, propostas de planificação, sugestões diversas, recursos e recomendações de avaliação. Tudo isto visa proporcionar aos professores elementos que lhes permitam vir a reunir um conjunto de condições necessárias a uma abordagem sustentada e atual do programa, em íntima articulação com o manual, de modo que os alunos procedam a uma aprendizagem que conduza a uma cidadania ativa.

Estudio de factibilidad para la creación de una empresa especializada en investigación de mercados, ubicada en la ciudad de Quito.

Diversas organizaciones tanto privadas como públicas en todo el mundo acuden a los servicios de asesoría como la capacitación, estudios de mercado, estudios de factibilidad, estudios sociológicos y de opinión pública, diseño de proceso y sistemas, reingeniería, asesoramiento técnico y más servicios profesionales. Ecuador no puede ser la excepción debido a los diversos problemas, cambios y sucesos que deben afrontar día a día las empresas como: la globalización, las competencias, clientes cada vez más exigentes, avances tecnológicos, permutas en la legislación, variaciones demográficas, tratados y acuerdos comerciales, entre otros. La mayor parte de las empresas que recuren a un servicio de asesoría lo hacen cuando una determinada situación se considera insatisfactoria y/o susceptible de mejora; por ende, la asistencia presentada por el asesor debe contribuir al mejoramiento de la calidad en los procesos de dirección y gestión de la empresa.

Estudio de factibilidad para la creación de un Centro de capacitación empresarial en la ciudad de Quito

El presente documento explica como está compuesto el estudio de factibilidad para la creación de un centro de capacitación empresarial en la ciudad de Quito, primeramente se hizo un extracto de la importancia de los centros de capacitación empresarial, sus diversas aplicaciones, métodos utilizados, razones por qué se debe de capacitar el individuo, cómo se maneja la capacitación empresarial en la actualidad, métodos utilizados para la capacitación y por último el papel que tiene la capacitación empresarial dentro de la empresa y sus diversas aplicaciones en ésta. Para establecer nuestro segmento de mercado se realizó una amplia investigación en lo que respecta a los centros de capacitación en la ciudad de Quito, de esta manera se pudo establecer a qué personas íbamos a enfocarnos en lo que respecta a la oferta, por otra parte la demanda fue investigada con profundidad con la finalidad de acercarnos a datos lo más confiables posibles. En cuanto tiene que ver al estudio técnico, éste se centró en establecer los más pertinentes recursos, tanto tecnológicos, humanos y de capital, que fueran los más convenientes para el desarrollo del Centro, pero siempre enfocados a mantener altos niveles de calidad para obtener instalaciones confortables, capacitadores entrenados y con una buena experiencia y métodos de marketing muy confiables como el Servqual y Servucción.

Estudio para el mejoramiento de seis centrales de climatización de la sala de tejedura de 4161.62 m2, que cuenta con 76 telares para tejido plano, de la fábrica textil "La Internacional"

En el área textil, sea esta tejeduría, hilandería, tintorería, o acabados de tela intervienen una infinidad de factores externos, relacionados con el tipo de material a procesar, se necesita tener un control riguroso de los factores externos tales como humedad relativa (HR) y temperatura, por lo que en el diseño de un área específica de cualquier planta textil, en este caso tejeduría, se invierte mucho dinero para garantizar condiciones estables de climatización tanto para el proceso como para el recurso humano.

Representaciones del cuerpo, discursos e identidad del pueblo afroecuatoriano.

En el territorio ancestral del Valle del Chota – La Concepción y Salinas, enclavado en el límite político administrativo de las provincias de Imbabura y Carchi, al norte del ecuador, habita una población de origen africano que fue introducida por la orden de Loyola a finales del siglo XVI en condición de esclavizado, destinados al cultivo de caña de azúcar, a quienes se les negó su condición humana. Para justifica la explotación económica en el ejercicio de la colonialidad del poder, los esclavizadores europeos, blancos, católicos y “Civilizados” se sustentaron en la falsa creencia etnocéntrica que la moral, los valores, la tecnología, la historia y la cultura de ellos, eran superiores a las de las personas esclavizadas. Desde entonces la sociedad ecuatoriana ha desconocido nuestros aportes económicos, sociales, culturales, religiosos y militares al país y, hasta hace poco, en la práctica, nuestros derechos de ciudadanía. Esto se conoce como racismo estructural.

Geologia - Guia do Professor - 12.º ano

É hoje amplamente reconhecida a relevância do papel desempenhado pelo professor de ciências no processo de ensino das temáticas científicas curriculares, tendo em vista a desejada aprendizagem dos alunos. Sem retirar a centralidade que estes têm no contexto da sua própria aprendizagem, os professores terão de procurar desenvolver competências didáticas que lhes permitam conceber, aplicar e avaliar estratégias capazes de reunir as melhores condições para os seus alunos aprenderem. Para este processo, assumidamente complexo, contribuem fontes diversas. Por exemplo, indicadores que resultam dos muitos estudos efetuados no âmbito da didática das ciências são deveras importantes. Destacam-se aqueles que se relacionam com a problematização, o trabalho prático, a história da ciência, a própria elaboração do conhecimento científico, a relação dos conceitos curriculares com os contextos social e tecnológico. Um bom nível dos saberes construídos pelos professores relativos aos temas curriculares a abordar é, igualmente, muito necessário. Um desempenho docente de qualidade está, também, articulado com uma atitude de disponibilidade do professor, quer para refletir sobre a sua própria experiência letiva, quer para a discutir e partilhar com outros colegas. Compreende-se, assim, que tenha havido a preocupação em elaborar o Guia do Professor, especialmente dirigido aos docentes responsáveis pela lecionação das temáticas de Geologia, do 12º ano. O documento, constituído por duas partes, tem a finalidade de ajudar os professores, de preferência trabalhando em cooperação com outros colegas, a pensar o desenvolvimento de estratégias de ensino adequadas às necessidades dos alunos timorenses do século XXI. Na primeira parte, são fundamentadas as orientações metodológicas, as quais incluem referências à educação para o desenvolvimento sustentável, algumas sugestões didáticas e observações relativas à avaliação das aprendizagens. Pretende-se que os professores desenvolvam um quadro de referência, o qual será importante para justificar algumas das decisões que terão de tomar na preparação das suas aulas e, depois, em situação de ato de ensino na sala de aula. Na segunda parte, são apresentados para cada uma das três Unidades Temáticas que integram o programa de Geologia do 12º ano, mapas de conceitos, propostas de planificação, sugestões diversas, recursos e recomendações de avaliação. Tudo isto visa proporcionar aos professores elementos que lhes permitam vir a reunir um conjunto de condições necessárias a uma abordagem sustentada e atual do programa, em íntima articulação com o manual, de modo que os alunos procedam a uma aprendizagem que conduza a uma cidadania ativa.