Immobilizing bacteriorhodopsin on a single electron transistor

As awareness of potential human and environmental impacts from toxins has increased, so has the development of innovative sensors. Bacteriorhodopsin (bR) is a light activated proton pump contained in the purple membrane (PM) of the bacteria Halobacterium salinarum. Bacteriorhodopsin is a robust protein which can function in both wet and dry states and can withstand extreme environmental conditions. A single electron transistor(SET) is a nano-scale device that exploits the quantum mechanical properties of electrons to switch on and off. SETs have tremendous potential in practical applications due to their size, ultra low power requirements, and electrometer-like sensitivity. The main goal of this research was to create a bionanohybrid device by integrating bR with a SET device. This was achieved by a multidisciplinary approach. The SET devices were created by a combination of sputtering, photolithography, and focused ion beam machining. The bionanomaterial bacteriorhodopsin was created through oxidative fermentation and a series of transmembrane purification processes. The bR was then integrated with the SET by electrophoretic deposition, creating a bionanohybrid device. The bionanohybrid device was then characterized using a semiconductor parametric analyzer. Characterization demonstrated that the bR modulated the operational characteristics of the SET when bR was activated with light within its absorbance spectrum. To effectively integrate bacteriorhodopsin with microelectromechanical systems (MEMS) and nanoelectromechanical systems (NEMS), it is critical to know the electrical properties of the material and to understand how it will affect the functionality of the device. Tests were performed on dried films of bR to determine if there is a relationship between inductance, capacitance, and resistance (LCR) measurements and orientation, light-on/off, frequency, and time. The results indicated that the LCR measurements of the bR depended on the thickness and area of the film, but not on the orientation, as with other biological materials such as muscle. However, there was a transient LCR response for both oriented and unoriented bR which depended on light intensity. From the impedance measurements an empirical model was suggested for the bionanohybrid device. The empirical model is based on the dominant electrical characteristics of the bR which were the parallel capacitance and resistance. The empirical model suggests that it is possible to integrate bR with a SET without influencing its functional characteristics.

Alternative splicing of pre-mRNA in cancer: focus on G protein-coupled peptide hormone receptors

Through alternative splicing, multiple different transcripts can be generated from a single gene. Alternative splicing represents an important molecular mechanism of gene regulation in physiological processes such as developmental programming as well as in disease. In cancer, splicing is significantly altered. Tumors express a different collection of alternative spliceoforms than normal tissues. Many tumor-associated splice variants arise from genes with an established role in carcinogenesis or tumor progression, and their functions can be oncogenic. This raises the possibility that products of alternative splicing play a pathogenic role in cancer. Moreover, cancer-associated spliceoforms represent potential diagnostic biomarkers and therapeutic targets. G protein-coupled peptide hormone receptors provide a good illustration of alternative splicing in cancer. The wild-type forms of these receptors have long been known to be expressed in cancer and to modulate tumor cell functions. They are also recognized as attractive clinical targets. Recently, splice variants of these receptors have been increasingly identified in various types of cancer. In particular, alternative cholecystokinin type 2, secretin, and growth hormone-releasing hormone receptor spliceoforms are expressed in tumors. Peptide hormone receptor splice variants can fundamentally differ from their wild-type receptor counterparts in pharmacological and functional characteristics, in their distribution in normal and malignant tissues, and in their potential use for clinical applications.

Intestinal macrophages: differentiation and involvement in intestinal immunopathologies

Intestinal macrophages, preferentially located in the subepithelial lamina propria, represent the largest pool of tissue macrophages in humans. As an adaptation to the local antigen- and bacteria-rich environment, intestinal macrophages exhibit several distinct phenotypic and functional characteristics. Notably, microbe-associated molecular pattern receptors, including the lipopolysaccharide (LPS) receptors CD14 and TLR4, and also the Fc receptors for IgA and IgG are absent on most intestinal macrophages under homeostatic conditions. Moreover, while macrophages in the intestinal mucosa are refractory to the induction of proinflammatory cytokine secretion, they still display potent phagocytic activity. These adaptations allow intestinal macrophages to comply with their main task, i.e., the efficient removal of microbes while maintaining local tissue homeostasis. In this paper, we review recent findings on the functional differentiation of monocyte subsets into distinct macrophage populations and on the phenotypic and functional adaptations that have evolved in intestinal macrophages in response to their antigen-rich environment. Furthermore, the involvement of intestinal macrophages in the pathogenesis of celiac disease and inflammatory bowel diseases is discussed.

(Table 1) Biomass (as energy) of different elements of the planktonic community in the Equatorial Pacific in the 0-150 m layer

An analysis was made of composition and content of nutrients, salts, particulate and dissolved organic matter, and various plankton groups in a series of samples collected by a 140-liter sampling bottle to depth up to 150 m at 4 equatorial stations between 97° and 154°W. Large and small phytoplankton, bacteria (aggregated and dispersed), heterotrophic flagellates, infusorians, radiolarians, foraminifers, fine filter-feeders, small and large, mostly herbivorous copepods, cyclopoids, predatory calanoids, and other predators were investigated separately. Trophic relations between these elements are established from personal and published data, and rate of their metabolism and some other physiological parameters are determined. Such functional characteristics as extent of satisfaction of food requirements of organisms belonging to various trophic groups, intensity of trophic relations, balance between production and consumption by individual elements of the community, ecological efficiency, and net and specific production of the groups distinguished, of individual trophic levels, of total zooplankton, and of the community as a whole are calculated. Variations of these characteristics along the equator with decreasing upwelling intensity are examined and their possible causes and mechanisms are discussed.

Effects of elevated CO2 and temperature on an intertidal meiobenthic community

In the near future, the marine environment is likely to be subjected to simultaneous increases in temperature and decreased pH. The potential effects of these changes on intertidal, meiofaunal assemblages were investigated using a mesocosm experiment. Artificial Substrate Units containing meiofauna from the extreme low intertidal zone were exposed for 60 days to eight experimental treatments (four replicates for each treatment) comprising four pH levels: 8.0 (ambient control), 7.7 & 7.3 (predicted changes associated with ocean acidification), and 6.7 (CO2 point-source leakage from geological storage), crossed with two temperatures: 12 °C (ambient control) and 16 °C (predicted). Community structure, measured using major meiofauna taxa was significantly affected by pH and temperature. Copepods and copepodites showed the greatest decline in abundance in response to low pH and elevated temperature. Nematodes increased in abundance in response to low pH and temperature rise, possibly caused by decreased predation and competition for food owing to the declining macrofauna density. Nematode species composition changed significantly between the different treatments, and was affected by both seawater acidification and warming. Estimated nematode species diversity, species evenness, and the maturity index, were substantially lower at 16 °C, whereas trophic diversity was slightly higher at 16 °C except at pH 6.7. This study has demonstrated that the combination of elevated levels of CO2 and ocean warming may have substantial effects on structural and functional characteristics of meiofaunal and nematode communities, and that single stressor experiments are unlikely to encompass the complexity of abiotic and biotic interactions. At the same time, ecological interactions may lead to complex community responses to pH and temperature changes in the interstitial environment.

Neighborhood mobility: A Long-Gone Pardigm? Approaching proximity and land use mix using data from home based surveys.

El WCTR es un congreso de reconocido prestigio internacional en el ámbito de la investigación del transporte y aunque las actas publicadas están en formato digital y sin ISSN ni ISBN, lo consideramos lo suficientemente importante como para que se considere en los indicadores. Policies trying to increase walking within urban mobility modal split usually highlight the importance of the functional patterns and the environmental quality of the urban space as major drivers of citizens modal choices. Functional characteristics would be mainly associated to an appropriate mix of land uses within neighbourhoods, whereas environmental quality would be associated to the characteristics of urban spaces. The purpose of this research is threefold: first, to identify relevant proxy indicators, which could characterize pedestrian-friendly land use mix and environmental quality. Second, to assess, for both traits, existing disparities among neighbourhoods in a major metropolitan area. And finally, to explore the association between both indicators and children mobility patterns: according to their built environment, which neighbourhoods have a greater proportion of children and, how is their mobility? Using data from the 2004 household mobility survey in the 128 neighbourhoods of the municipality of Madrid, this paper concludes that potentially favourable conditions at the neighbourhood level seem to have only a modest influence in,mobility patterns , in terms of both, selection of closer destinations and a higher share of walking within modal split. The citys policy choices, with intensive investment in road and public transport infrastructure may explain why short-distance mobility is not as important as it could have been expected in those neighbourhoods with more pedestrian-friendly conditions. The metropolitan transport system is providing mobility conditions, which make far-away destinations attractive to most citizens.

Neighbourhood mobility: a longgone Paradigm? Approaching "proximity" and "land use mix" using data from home-based surveys

Policies trying to increase walking within urban mobility modal split usually highlight the importance of the functional patterns and the environmental quality of the urban space as major drivers of citizens modal choices. Functional characteristics would be mainly associated to an appropriate mix of land uses within neighbourhoods, whereas environmental quality would be associated to the characteristics of urban spaces. The purpose of this research is threefold: first, to identify relevant proxy indicators, which could characterize pedestrian-friendly land use mix and environmental quality. Second, to assess, for both traits, existing disparities among neighbourhoods in a major metropolitan area. And finally, to explore the association between both indicators and children mobility patterns: according to their built environment, which neighbourhoods have a greater proportion of children and, how is their mobility? Using data from the 2004 household mobility survey in the 128 neighbourhoods of the municipality of Madrid, this paper concludes that potentially favourable conditions at the neighbourhood level seem to have only a modest influence in,mobility patterns , in terms of both, selection of closer destinations and a higher share of walking within modal split. The city s policy choices, with intensive investment in road and public transport infrastructure may explain why short-distance mobility is not as important as it could have been expected in those neighbourhoods with more pedestrian-friendly conditions. The metropolitan transport system is providing mobility conditions, which make far-away destinations attractive to most citizens.

Integración en la plataforma 4mobile de mecanismos de atención en redes sociales

El presente texto se ha desarrollado como Proyecto Fin de Grado en la Escuela Técnica Superior de Ingeniería y Sistemas de Telecomunicación de la Universidad Politécnica de Madrid en colaboración con la Consultora Tecnológica everis. El mismo tiene como objetivo realizar un estudio de los requisitos necesarios para poder desplegar un servicio que permita ofrecer a las Operadoras Móviles Virtuales (OMV) soporte técnico a sus clientes a través de las redes sociales. Este módulo establecerá una nueva vía de comunicación entre el consumidor y el cliente de la OMV con la propia OMV, el cual se caracterizará por una alta accesibilidad y una gran rapidez en el servicio permitiendo de este modo, una mayor satisfacción del cliente con su operador móvil y por tanto de una nueva vía para conseguir la fidelización del mismo y la captación de nuevos clientes. Para ello, este proyecto se lleva a cabo sobre el sistema de información 4mobile el cual es comercializado por la mencionada Consultora Tecnológica. Este sistema consiste en una plataforma web la cual permite cubrir todos los procesos de negocio comunes que un OMV necesita gestionar. Es por ello, que el mencionado estudio se centra en la evaluación de los aspectos necesarios para la integración de un módulo de estas características dentro de la plataforma de 4mobile. Este módulo, estará basado en una herramienta software que permitirá gestionar el ciclo de vida completo del comentario realizado por el cliente a través de una red social, desde que es publicado por el cliente, hasta que se haya respondido al mismo y su solución sea considerada como satisfactoria por el cliente. Por ello, y de cara a definir correctamente esta herramienta, será necesario un detallado análisis el cual recoja diversos aspectos y que a lo largo del texto, será denominado como Plan de Marketing de Medios Sociales (PMMS). Estos aspectos versarán tanto de las necesidades tecnológicas para su mencionada integración, como de la serie de características funcionales que una solución basada en servicio técnico a través de las redes sociales deberá poseer con el objetivo de ofrecer un servicio técnico de calidad. Finalmente, estas funcionalidades y necesidades tecnológicas se expondrán en forma de propuesta a everis para su integración en la plataforma 4mobile junto con un análisis de diseño a alto nivel software de la solución a desarrollar. ABSTRACT. This text has been developed as Final Degree Project in the Escuela Técnica Superior de Ingeniería y Sistemas de Telecomunicación de la Universidad Politécnica de Madrid in collaboration with the Technology Consultant everis, aims to conduct a study of how to provide technical support through social networks and the evaluation of the integration of a social support module within the system platform 4mobile, which cover all business processes that need to manage a Mobile Virtual Network Operator (MVNO), which is marketed by above-mentioned consultant . This module will establish a new communication channel between the consumer and the client with the OMV and itself, which is characterized by high accessibility and great fast service. Thus will allow a higher customer satisfaction with him service and thus a new way to get the same loyalty and attract new customers To this end, this project is performed on the information system 4mobile which is marketed by the consulting part thereof everis. This system is a platform to cover all business processes that need to manage an MVNO. Therefore, the mentioned study focuses on the evaluation of the elements necessary for integrating module these features within the platform 4mobile. This module will be based on a software tool for managing the entire lifecycle of the comment made by the customer via social networking, from the moment it is published, until it has been respond and their solution is considered satisfactory for the customer. So, in order to properly define this tool, a detailed analysis which will be necessary to collect various aspects and throughout the text, it will be referred to as Plan of Social Media Marketing (PMMS). These aspects will be address both: the technological needs for the mentioned integration, and the number of functional characteristics-based service through social networking solution, as this must in order to provide a quality service Finally, these goals and requirements will be discussed as everis offer for integration into the platform 4mobile analysis along with a high level of software design to develop the solution.

Optimization and evaluation in space conditions of multi-GHz optical modulators

Among the different optical modulator technologies available such as polymer, III-V semiconductors, Silicon, the well-known Lithium Niobate (LN) offers the best trade-off in terms of performances, ease of use, and power handling capability [1-9]. The LN technology is still widely deployed within the current high data rate fibre optic communications networks. This technology is also the most mature and guarantees the reliability which is required for space applications [9].In or der to fulfil the target specifications of opto-microwave payloads, an optimization of the design of a Mach-Zehnder (MZ) modulator working at the 1500nm telecom wavelength was performed in the frame of the ESA-ARTES "Multi GigaHertz Optical Modulator" (MGOM) project in order to reach ultra-low optical insertion loss and low effective driving voltage in the Ka band. The selected modulator configuration was the X-cut crystal orientation, associated to high stability Titanium in-diffusion process for the optical waveguide. Starting from an initial modulator configuration exhibiting 9 V drive voltage @ 30 GHz, a complete redesign of the coplanar microwave electrodes was carried out in order to reach a 6 V drive voltage @ 30GHz version. This redesign was associated to an optimization of the interaction between the optical waveguide and the electrodes. Following the optimisation steps, an evaluation program was applied on a lot of 8 identical modulators. A full characterisation was carried out to compare performances, showing small variations between the initial and final functional characteristics. In parallel, two similar modulators were submitted to both gamma (10-100 krad) and proton irradiation (10.109 p/cm²) with minor performance degradation.

Characterization of human cardiac Na+ channel mutations in the congenital long QT syndrome

The congenital long QT syndrome (LQTS) is an inherited disorder characterized by a prolonged cardiac action potential. This delay in cellular repolarization can lead to potentially fatal arrhythmias. One form of LQTS (LQT3) has been linked to the human cardiac voltage-gated sodium channel gene (SCN5A). Three distinct mutations have been identified in the sodium channel gene. The biophysical and functional characteristics of each of these mutant channels were determined by heterologous expression of a recombinant human heart sodium channel in a mammalian cell line. Each mutation caused a sustained, non-inactivating sodium current amounting to a few percent of the peak inward sodium current, observable during long (>50 msec) depolarizations. The voltage dependence and rate of inactivation were altered, and the rate of recovery from inactivation was changed compared with wild-type channels. These mutations in diverse regions of the ion channel protein, all produced a common defect in channel gating that can cause the long QT phenotype. The sustained inward current caused by these mutations will prolong the action potential. Furthermore, they may create conditions that promote arrhythmias due to prolonged depolarization and the altered recovery from inactivation. These results provide insights for successful intervention in the disease.

A cell surface receptor defined by a mAb mediates a unique type of cell death similar to oncosis

Cell death is mediated by distinct pathways including apoptosis and oncosis in response to various death signals. To characterize molecules involved in cell death, a panel of mAbs was raised by immunizing mice with apoptotic cells. One of these antibodies, designated anti-Porimin (for pro-oncosis receptor inducing membrane injury), was found to directly induce a unique type of cell death in Jurkat cells. Anti-Porimin defines a 110-kDa cell surface receptor on Jurkat cells. Functionally, anti-Porimin alone rapidly mediates pore formation on the plasma membrane and induces cell death without participation of complement. Both the cellular expression and functional characteristics of the Porimin antigen indicate that it is distinct from the CD95 (Fas/Apo-1) and other cell receptors known to induce apoptosis. Anti-Porimin-mediated cell death was preceded by cell aggregation, formation of plasma membrane pores, and the appearance of membrane blebs. More important, these cells show neither DNA fragmentation nor apoptotic bodies, but display lethal damage of the cell membrane. Cell death by anti-Porimin is distinct from complement-dependent cytolysis or complement-independent apoptosis but is similar to that described for oncosis, a form of cell death accompanied by the membrane damage followed by karyolysis. The induction of cell death by anti-Porimin may represent a unique cell surface receptor-mediated pathway of cell death in the human lymphoid system.

Endothelial cell death, angiogenesis, and microvascular function after castration in an androgen-dependent tumor: Role of vascular endothelial growth factor

The sequence of events that leads to tumor vessel regression and the functional characteristics of these vessels during hormone–ablation therapy are not known. This is because of the lack of an appropriate animal model and monitoring technology. By using in vivo microscopy and in situ molecular analysis of the androgen-dependent Shionogi carcinoma grown in severe combined immunodeficient mice, we show that castration of these mice leads to tumor regression and a concomitant decrease in vascular endothelial growth factor (VEGF) expression. Androgen withdrawal is known to induce apoptosis in Shionogi tumor cells. Surprisingly, tumor endothelial cells begin to undergo apoptosis before neoplastic cells, and rarefaction of tumor vessels precedes the decrease in tumor size. The regressing vessels begin to exhibit normal phenotype, i.e., lower diameter, tortuosity, vascular permeability, and leukocyte adhesion. Two weeks after castration, a second wave of angiogenesis and tumor growth begins with a concomitant increase in VEGF expression. Because human tumors often relapse following hormone–ablation therapy, our data suggest that these patients may benefit from combined anti-VEGF therapy.

A family of genes required for maintenance of cell wall integrity and for the stress response in Saccharomyces cerevisiae

The PKC1–MPK1 pathway in yeast functions in the maintenance of cell wall integrity and in the stress response. We have identified a family of genes that are putative regulators of this pathway. WSC1, WSC2, and WSC3 encode predicted integral membrane proteins with a conserved cysteine motif and a WSC1–green fluorescence protein fusion protein localizes to the plasma membrane. Deletion of WSC results in phenotypes similar to mutants in the PKC1–MPK1 pathway and an increase in the activity of MPK1 upon a mild heat treatment is impaired in a wscΔ mutant. Genetic analysis places the function of WSC upstream of PKC1, suggesting that they play a role in its activation. We also find a genetic interaction between WSC and the RAS–cAMP pathway. The RAS–cAMP pathway is required for cell cycle progression and for the heat shock response. Overexpression of WSC suppresses the heat shock sensitivity of a strain in which RAS is hyperactivated and the heat shock sensitivity of a wscΔ strain is rescued by deletion of RAS2. The functional characteristics and cellular localization of WSC suggest that they may mediate intracellular responses to environmental stress in yeast.

Expression of γ-aminobutyric acid ρ1 and ρ1Δ450 as gene fusions with the green fluorescent protein

The functional characteristics and cellular localization of the γaminobutyric acid (GABA) ρ1 receptor and its nonfunctional isoform ρ1Δ450 were investigated by expressing them as gene fusions with the enhanced version of the green fluorescent protein (GFP). Oocytes injected with ρ1-GFP had receptors that gated chloride channels when activated by GABA. The functional characteristics of these receptors were the same as for those of wild-type ρ1 receptors. Fluorescence, because of the chimeric receptors expressed, was over the whole oocyte but was more intense near the cell surface and more abundant in the animal hemisphere. Similar to the wild type, ρ1Δ450-GFP did not lead to the expression of functional GABA receptors, and injected oocytes failed to generate currents even after exposure to high concentrations of GABA. Nonetheless, the fluorescence displayed by oocytes expressing ρ1Δ450-GFP was distributed similarly to that of ρ1-GFP. Mammalian cells transfected with the ρ1-GFP or ρ1Δ450-GFP constructs showed mostly intracellularly distributed fluorescence in confocal microscope images. A sparse localization of fluorescence was observed in the plasma membrane regardless of the cell line used. We conclude that ρ1Δ450 is expressed and transported close to, and perhaps incorporated into, the plasma membrane. Thus, ρ1- and ρ1Δ450-GFP fusions provide a powerful tool to visualize the traffic of GABA type C receptors.

The purification and characterization of fetal liver hematopoietic stem cells.

Thy-1loSca-1+Lin-Mac-1+CD4- cells have been isolated from the livers of C57BL-Thy-1.1 fetuses. This population appears to be an essentially pure population of hematopoietic stem cells (HSC), in that injection of only six cells into lethally irradiated adult recipients yields a limit dilution frequency of donor cell-reconstituted mice. Sixty-seven to 77% of clones in this population exhibit long-term multilineage progenitor activity. This population appears to include all long-term multilineage reconstituting progenitors in the fetal liver. A high proportion of cells are in cycle, and the absolute number of cells in this population doubles daily in the fetal liver until 14.5 days postcoitum. At 15.5 days postcoitum, the frequency of this population falls dramatically. Long-term reconstituting HSC clones from the fetal liver give rise to higher levels of reconstitution in lethally irradiated mice than long-term reconstituting HSC from the bone marrow. The precise phenotypic and functional characteristics of HSC vary according to tissue and time during ontogeny.