Expression and characterization of the Trypanosoma cruzi dihydrofolate reductase domain

We have cloned and expressed in Escherichia coli a 702-base pair gene coding for the dihydrofolate reductase (DHFR) domain of the bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) from Trypanosoma cruzi. The DHFR domain was purified to homogeneity by methotrexate-Sepharose chromatography followed by an anion-exchange chromatography step in a mono Q column, and displayed a single 27-kDa band on SDS-PAGE. Gel filtration showed that the catalytic domain was expressed as a monomer. Kinetic parameters were similar to those reported for the wild-type bifunctional enzyme with Km values of 0.75 microM for dihydrofolate and 16 microM for NADPH and a kcat value of 16.5 s-1. T. cruzi DHFR is poorly inhibited by trimethoprim and pyrimethamine and the inhibition constants were always lower for the bifunctional enzyme. The binding of methotrexate was characteristic of a class of inhibitors that form an initial complex which isomerizes slowly to a tighter complex and are referred to as 'slow, tight-binding' inhibitors. While the slow-binding step of inhibition was apparently unaffected in the individually expressed DHFR domain, the overall inhibition constant was two-fold higher as a consequence of the superior inhibition constant value obtained for the initial inhibitory complex

Influencia del tiempo de uso de las tecnologías de información y comunicación y subprogramas sobre las habilidades informacionales en estudiantes de la Universidad del Zulia

The purpose of this research was to determine the influence of the time of use of technologies of information and communication and related subprograms in the informational skills of students at the University of the Zulia. The research was developed under a descriptive, field based methodology, with a non-experimental design of the trans-sectional type. The population was made up of 95 students in the economic and social sciences program at the LUZ COL. Nucleus. The sample was obtained by means of the Shiffer formula and made up of a group of 52 students in the subprogram of accounting, 15 students of economy, and 18 students of administration. A random sampling stratified by subprograms was used. For the compilation of information 3 instruments were designed, the first one with 8 simple questions, the second one with 29 questions and a Lickert’s scale with four alternatives, and the third one was an observation sheet. The validation of the instrument was made by three experts in the area and there was no the need to realize a reliability test of the instrument provided that they were questionnaires. The analysis of the information was realized by the statistical program SPSS Version 10, in which tests were realized for the calculations of arithmetic averages and standard deviation, as well as analysis of variance, studies of homogeneity and Tukey comparison tests in order to determine the value of the variables in the study. The results obtained showed an influence of the time of use of TIC across the dimension of frequency of hours per week,and informational skills, specifically in the aspects of access and communication of information, and also they revealed that the subprograms do not influence these skills, where students in the program possess high use of these informational skills.El propósito de esta investigación fue determinar la influencia del tiempo de uso de las tecnologías de información y comunicación y subprogramas sobre las habilidades informacionales en estudiantes de la Universidad del Zulia. La investigación fue desarrollada bajo la metodología descriptiva, de campo, con un diseño no experimental del tipo transeccional descriptivo. La población estuvo conformada por 95 estudiantes del programa de ciencias económicas y sociales del núcleo COL de LUZ. La muestra se obtuvo mediante la fórmula de Shiffer constituyéndose por una muestra de 52 estudiantes del subprograma de contaduría, 15 de economía y 18 de administración. Se utilizó un muestreo aleatorio estratificado por subprogramas. Para la recolección de datos se diseñarón 3 instrumentos, dos cuestionarios y una hoja de observación con su respectiva validación a cargo de tres expertos. El análisis de los datos se realizó a través del programa estadístico SPSS Versión 10, en el cual se realizaron pruebas para los cálculos de media aritmética y desviación estándar, además del análisis de la varianza, estudios de Homogeneidad y pruebas de comparación de Tukey y robustas, con el fin de determinar el valor de las variables en el estudio. Los resultados obtenidos mostraron una influencia del tiempo en el uso de las TIC a través de su dimensión frecuencia de horas a la semana sobre las habilidades informacionales, específicamente en los aspectos de acceso y comunicación de la información, también revelaron que los subprogramas no influyen sobre estas destrezas, donde los estudiantes del programa poseen un uso Alto de las habilidades Informacionale

Quantum time machine

The continuation of Misner space into the Euclidean region is seen to imply the topological restriction that the period of the closed spatial direction becomes time-dependent. This restriction results in a modified Lorentzian Misner space in which the renormalized stress-energy tensor for quantized complex massless scalar fields becomes regular everywhere, even on the chronology horizon. A quantum-mechanically stable time machine with just the sub-microscopic size may then be constructed out of the modified Misner space, for which the semiclassical Hawking's chronology protection conjecture is no longer an obstruction.Comment: 6 pages, RevTe

Wormholes and Ringholes in a Dark-Energy Universe

The effects that the present accelerating expansion of the universe has on the size and shape of Lorentzian wormholes and ringholes are considered. It is shown that, quite similarly to how it occurs for inflating wormholes, relative to the initial embedding-space coordinate system, whereas the shape of the considered holes is always preserved with time, their size is driven by the expansion to increase by a factor which is proportional to the scale factor of the universe. In the case that dark energy is phantom energy, which is not excluded by present constraints on the dark-energy equation of state, that size increase with time becomes quite more remarkable, and a rather speculative scenario is here presented where the big rip can be circumvented by future advanced civilizations by utilizing sufficiently grown up wormholes and ringholes as time machines that shortcut the big-rip singularity.Comment: 11 pages, RevTex, to appear in Phys. Rev.

The NOBEL2 approach to resilience in future transport networks

IST project NOBEL2 results on resilience strategies for next-generation optical transport networks are presented, paving the way towards cost-effective, scalable and easy-to-maintain multi-service network architectures.Postprint (published version

Cloning and expression of the dihydrofolate reductase-thymidylate synthase gene from Trypanosoma cruzi

We have cloned, sequenced and expressed the Trypanosoma cruzi gene encoding the bifunctional protein dihydrofolate reductase-thymidylate synthase (DHFR-TS). The strategy followed for the isolation of positive clones from a genomic library was based on the construction of a probe by the amplification of highly conserved sequences of the TS domain by the polymerase chain reaction. Translation of the open reading frame of 1563 bp yields a polypeptide of 521 amino acids with a molecular mass of 58829 Da. For heterologous expression of T. cruzi DHFR-TS in Escherichia coli, the entire coding sequence was amplified by polymerase chain reaction and cloned into the plasmid vector pKK223.3. The presence of catalytically active DHFR-TS was demonstrated by complementation of the Thy- E. coli strain chi 2913 and the DHFR- Thy- E. coli strain PA414. The gene is expressed as an active protein which constitutes approximately 2% of the total cell soluble protein. Recombinant bifunctional enzyme and the DHFR domain have been purified by methotrexate-Sepharose chromatography to yield 1-2 mg of active DHFR-TS per litre of culture. Southern and electrophoretic analyses using the coding sequence as probe indicated that the T. cruzi enzyme is encoded by a single copy gene which maps to two bands of approximately 990 kb and 1047 kb. It appears that T. cruzi is diploid for the DHFR-TS gene which is located on two different-sized homologous chromosomes

Ringholes and closed timelike curves

It is shown that in a classical spacetime with multiply connected space slices having the topology of a torus, closed timelike curves are also formed. We call these spacetime ringholes. Two regions on the torus surface can be distinguished which are separated by angular horizons. On one of such regions (that which surrounds the maximum circumference of the torus) everything happens like in spherical wormholes, but the other region (the rest of the torus surface), while still possessing a chronology horizon and non-chronal region, behaves like a coverging, rather than diverging, lens and corresponds to an energy density which is always positive for large speeds at or near the throat. It is speculated that a ringhole could be converted into a time machine to perform time travels by an observer who would never encounter any matter that violates the classical averaged weak energy condition. Based on a calculation of vacuum fluctuations, it is also seen that the angular horizons can prevent the emergence of quantum instabilities near the throat.Comment: 11 pages, RevTex, 4 figures available upon reques

Ocean Circulation over Formigas and Ormonde Seamounts

Seamounts constitute an obstacle to the free ocean flow, modifying the patter of circulation. As a result of these alterations, a variety of hydrodynamical processes and phenomena may take place in seamounts, among others, Taylor columns/caps. These oceanographic effects may turn seamounts into very productive ecosystems with high biodiversity. Under these conditions seamounts provide a particularly good environment for the settle of some organisms, acting as stepping stones and contributing to its dispersal. In this study, we verify if these oceanographic effects explain the presence of cold-water corals of Mediterranean origin in the Atlantic. To achieve this, three seamounts in the path of the Mediterranean Outflow Water (MOW) through the Eastern North Atlantic were selected: the Gazul mud volcano, and the Ormonde and Formigas seamounts. In order to determine the hydrographic and dynamical conditions in each one of the three locations, CTD, LADPC and biochemical observations were carried out. Taylor columns were not observed in any of the three sampled areas. Although we found suggestions of upwelling/downwelling systems, their effect was barely noticed in the circulation pattern. The oceanographic processes in those areas are more influenced by the vertical distribution of water masses, which determine the stability of the water column. Moreover, the high values of the Brunt-Väisälä frequency around the MOW halocline can lead to the formation of internal waves. These perturbations in the water column can enhance the vertical mixing, producing suspension, which, in turn, could affect the vertical distribution of cold-water corals

Orogenias paleozoicas en los Andes de Argentina y Chile y en la Península Antártica

Congreso Geológico Argentino (20º. 2017. San Miguel de Tucumán, Argentina). Simposio de Téctonica pre-andinaDurante el Neoproterozoico y Paleozoico, los Andes de Argentina y Chile, y desde fines del Paleozoico también la Península Antártica, formaron parte del margen SO de Gondwana. Durante este tiempo se acrecionaron a dicho margen varios fragmentos continentales de tamaño y aloctonía variable; denominados de N a S: Antofalla, Chi-Cu, Patagonia Occidental y Antártida Occidental. Estos fragmentos formaban parte de placas litosféricas, en ocasiones divididas en subplacas. La colisión de dichos fragmentos continentales con Gondwana y una última subducción bajo dicho margen, dieron lugar a 6 orogenias de extensión temporal y espacial limitada.Instituto Geológico y Minero de España, EspañaDepartamento de Geología, Universidad de Oviedo, EspañaUniversidad de Río Negro, ArgentinaServicio Geológico y Minero Argentino, ArgentinaInstituto De Bio y Geociencias Del NOA, Consejo Nacional de Investigaciones Científicas y Técnicas, ArgentinaInstituto De Bio y Geociencias Del NOA, Universidad Nacional de Salta, ArgentinaDepartamento de Geodinámica, Universidad del País Vasco, EspañaFacultad de Geología, Universidad de Barcelona, EspañaDepartamento de Geología, Universidad de Chile, ChileUniversidad Andrés Bello, ChileUnidad de Tectónica, Consejo Nacional de Investigaciones Científicas y Técnicas, ArgentinaFacultad de Geología, Universidad de Buenos Aires, ArgentinaÁrea de Geología, Universidad Rey Juan Carlos, EspañaUniversidad de Salta, ArgentinaInstituto de Investigación en Paleobiología y Geología, Universidad de Río Negro, ArgentinaInstituto de Investigación en Paleobiología y Geología, Consejo Nacional de Investigaciones Científicas y Técnicas, ArgentinaCentro de Investigaciones Geológicas, Universidad de La Plata, ArgentinaUniversidad de San Juan, ArgentinaPeer reviewe