963 resultados para Idaho National Engineering Laboratory.
Resumo:
The principal risks in the railway industry are mainly associated with collisions, derailments and level crossing accidents. An understanding of the nature of previous accidents on the railway network is required to identify potential causes and develop safety systems and deploy safety procedures. Risk assessment is a process for determining the risk magnitude to assist with decision-making. We propose a three-step methodology to predict the mean number of fatalities in railway accidents. The first is to predict the mean number of accidents by analyzing generalized linear models and selecting the one that best fits to the available historical data on the basis of goodness-offit statistics. The second is to compute the mean number of fatalities per accident and the third is to estimate the mean number of fatalities. The methodology is illustrated on the Spanish railway system. Statistical models accounting for annual and grouped data for the 1992-2009 time period have been analyzed. After identifying the models for broad and narrow gauges, we predicted mean number of accidents and the number of fatalities for the 2010-18 time period.
Resumo:
The engineer must have sufficient theoretical knowledge to be applied to solve specific problems, with the necessary capacity to simplify these approaches, and taking into account factors such as speed, simplicity, quality and economy. In Geology, its ultimate goal is the exploration of the history of the geological events through observation, deduction, reasoning and, in exceptional cases by the direct underground exploration or experimentation. Experimentation is very limited in Geology. Reproduction laboratory of certain phenomena or geological processes is difficult because both time and space become a large scale. For this reason, some Earth Sciences are in a nearly descriptive stage whereas others closest to the experimental, Geophysics and Geochemistry, have assimilated progress experienced by the physics and chemistry. Thus, Anglo-Saxon countries clearly separate Engineering Geology from Geological Engineering, i.e. Applied Geology to the Geological Engineering concepts. Although there is a big professional overlap, the first one corresponds to scientific approach, while the last one corresponds to a technological one. Applied Geology to Engineering could be defined as the Science and Applied Geology to the design, construction and performance of engineering infrastructures in and field geology discipline. There has been much discussion on the primacy of theory over practice. Today prevails the exaggeration of practice, but you get good workers and routine and mediocre teachers. This idea forgets too that teaching problem is a problem of right balance. The approach of the action lines on the European Higher Education Area (EHEA) framework provides for such balance. Applied Geology subject represents the first real contact with the physical environment with the practice profession and works. Besides, the situation of the topic in the first trace of Study Plans for many students implies the link to other subjects and topics of the career (tunnels, dams, groundwater, roads, etc). This work analyses in depth the justification of such practical trips. It shows the criteria and methods of planning and the result which manifests itself in pupils. Once practical trips experience developed, the objective work tries to know about results and changes on student’s motivation in learning perspective. This is done regardless of the outcome of their knowledge achievements assessed properly and they are not subject to such work. For this objective, it has been designed a survey about their motivation before and after trip. Survey was made by the Unidad Docente de Geología Aplicada of the Departamento de Ingeniería y Morfología del Terreno (Escuela Técnica Superior de Ingenieros de Caminos, Canales y Puertos, Universidad Politécnica de Madrid). It was completely anonymous. Its objective was to collect the opinion of the student as a key agent of learning and teaching of the subject. All the work takes place under new teaching/learning criteria approach at the European framework in Higher Education. The results are exceptionally good with 90% of student’s participation and with very high scores in a number of questions as the itineraries, teachers and visited places (range of 4.5 to 4.2 in a 5 points scale). The majority of students are very satisfied (average of 4.5 in a 5 points scale).
Resumo:
The purpose of this report is to build a model that represents, as best as possible, the seismic behavior of a pile cap bridge foundation by a nonlinear static (analysis) procedure. It will consist of a reproduction of a specimen already built in the laboratory. This model will carry out a pseudo static lateral and horizontal pushover test that will be applied onto the pile cap until the failure of the structure, the formation of a plastic hinge in the piles due to the horizontal deformation, occurs. The pushover test consists of increasing the horizontal load over the pile cap until the horizontal displacement wanted at the height of the pile cap is reached. The output of this model will be a Skeleton curve that will plot the lateral load (kN) over the displacement (m), so that the maximum movement the pile cap foundation can reach before its failure can be calculated. This failure will be achieved when the load at that specific shift is equal to 85% of the maximum. The pile cap foundation finite element model was based on pile cap built for a laboratory experiment already carried out by the Master student Deming Zhang at Tongji University. Two different pile caps were tested with a difference in height above the ground level. While one has 0:3m, the other rises 0:8m above the ground level. The computer model was calibrated using the experimental results. The pile cap foundation will be programmed in a finite element environment called OpenSees (Open System for Earthquake Engineering Simulation [28]). This environment is a free software developed by Berkeley University specialized, as it name says, in the study of earthquakes and its effects on structures. This specialization is the main reason why it is being used for building this model as it makes it possible to build any finite element model, and perform several analysis in order to get the results wanted. The development of OpenSees is sponsored by the Pacific Earthquake Engineering Research Center through the National Science Foundation engineering and education centers program. OpenSees uses Tcl language to program it, which is a language similar to C++.
Resumo:
Automatic Control Teaching in the new degree syllabus has reduced both, its contents and its implementation course, with regard to traditional engineering careers. On the other hand, where the qualification is not considered as automatic control specialist, it is required an adapted methodology to provide the minimum contents that the student needs to assimilate, even in the case that students do not perceive these contents as the most important in their future career. In this paper we present the contents of a small automatic course taught Naval Architecture and Marine Engineering Degrees at the School of Naval Engineering of the Polytechnic University of Madrid. We have included the contents covered using the proposed methodology which is based on practical work after lectures. Firstly, the students performed exercises by hand. Secondly, they solve the exercises using informatics support tools, and finally, they validate their previous results and their knowledge in the laboratory platforms.
Resumo:
En el Campus Sur de la Universidad Politécnica de Madrid se ha llevado a cabo un proyecto para obtener una caracterización del subsuelo mediante ensayos ReMi, en colaboración con el departamento de Geofísica del Instituto Geográfico Nacional. La técnica ReMi (Refraction Microtremor) permite, mediante ensayos geofísicos realizados localmente sobre el terreno,obtener los parámetros físicos del mismo, que resultan de especial interés en el ámbito de la ingeniería civil. Esta técnica se caracteriza por englobarse dentro de la sísmica pasiva, muy empleada en prospección geofísica y basada en la obtención del modelo subyacente de distribución de velocidades de propagación de la onda S en función de la profundidad, con la ventaja de aprovechar el ruido sísmico ambiental como fuente de energía. Fue desarrollada en el Laboratorio Sismológico de Nevada (EEUU) por Louie (2001), con el objetivo de presentar una técnica innovadora en la obtención de las velocidades de propagación de manera experimental. Presenta ciertas ventajas, como la observación directa de la dispersión de ondas superficiales,que da un buen resultado de la velocidad de onda S, siendo un método no invasivo, de bajo coste y buena resolución, aplicable en entornos urbanos o sensibles en los que tanto otras técnicas sismológicas como otras variedades de prospección presentan dificultades. La velocidad de propagación de la onda S en los 30 primeros metros VS30, es ampliamente reconocida como un parámetro equivalente válido para caracterizar geotécnicamente el subsuelo y se halla matemáticamente relacionada con la velocidad de propagación de las ondas superficiales a observar mediante la técnica ReMi. Su observación permite el análisis espectral de los registros adquiridos, obteniéndose un modelo representado por la curva de dispersión de cada emplazamiento, de modo que mediante una inversión se obtiene el modelo de velocidad de propagación en función de la profundidad. A través de estos modelos, pueden obtenerse otros parámetros de interés sismológico. Estos resultados se representan sobre mapas isométricos para obtener una relación espacial de los mismos, particularmente conocido como zonación sísmica. De este análisis se extrae que la VS30 promedio del Campus no es baja en exceso, correspondiéndose a posteriori con los resultados de amplificación sísmica, período fundamental de resonancia del lugar y profundidad del sustrato rocoso. En última instancia se comprueba que los valores de amplificación sísmica máxima y el período al cual se produce posiblemente coincidan con los períodos fundamentales de resonancia de algunos edificios del Campus. ABSTRACT In South Campus at Polytechnic University of Madrid, a project has been carried out to obtain a proper subsoil description by applying ReMi tests, in collaboration with the Department of Geophysics of the National Geographic Institute. Through geophysical tests conducted locally, the ReMi (Refraction Microtremor) technique allows to establish the physical parameters of soil, which are of special interest in the field of civil engineering. This technique is part of passive seismic methods, often used in geophysical prospecting. It focuses in obtaining the underlying model of propagation velocity distribution of the shear wave according to depth and has the advantage of being able to use seismic ambient noise as a source of energy. It was developed in the Nevada Seismological Laboratory (USA) by Louie (2001) as an innovative technique for obtaining propagation velocities experimentally. It has several other advantages, including the direct observation of the dispersion of surface waves, which allows to reliably measure S wave velocity. This is a non-invasive, low cost and good resolution method, which can be applied in urban or sensitive environments where other prospection methods present difficulties. The propagation velocity of shear waves in the first 30 meters Vs30 is widely recognized as a valid equivalent parameter to geotechnically characterize the subsurface. It is mathematically related to surface wave's velocity of propagation, which are to observe using REMI technique. Spectral analysis of acquired data sets up a model represented by the dispersion curve at each site, so that, using an inversion process, propagation velocity model in relation to depth is obtained. Through this models, other seismologically interesting parameters can be obtained. These results are represented on isometric maps in order to obtain a spatial relationship between them, a process which is known as seismic zonation. This analysis infers that Vs30 at South Campus is not alarmingly low , corresponding with subsequent results of seismic amplification, fundamental period of resonance of soil and depth of bedrock. Ultimately, it's found that calculated values of soil's fundamental periods at which maximum seismic amplification occurs, may possibly match fundamental periods of some Campus buildings.
Resumo:
Accreditation models in the international context mainly consider the evaluation of learning outcomes and the ability of programs (or higher education institutions) to achieve the educational objectives stated in their mission. However, it is not clear if these objectives and therefore their outcomes satisfy real national and regional needs, a critical point in engineering master's programs, especially in developing countries. The aim of this paper is to study the importance of the local relevancy evaluation of these programs and to analyze the main models of quality assurance and accreditation bodies of USA, Europe and Latin America, in order to ascertain whether the relevancy is evaluated or not. After a literature review, we found that in a free-market economic context and international education, the accreditation of master's programs follows an international accreditation model, and doesńt take in account in most cases criteria and indicators for local relevancy. It concludes that it is necessary both, international accreditation to ensure the effectiveness of the program (achievement of learning outcomes) and the national accreditation through which it could ensure local relevancy of programs, for which we are giving some indicators.
Resumo:
Ligand-specific molecular switches composed of RNA were created by coupling preexisting catalytic and receptor domains via structural bridges. Binding of ligand to the receptor triggers a conformational change within the bridge, and this structural reorganization dictates the activity of the adjoining ribozyme. The modular nature of these tripartite constructs makes possible the rapid construction of precision RNA molecular switches that trigger only in the presence of their corresponding ligand. By using similar enzyme engineering strategies, new RNA switches can be made to operate as designer molecular sensors or as a new class of genetic control elements.
Resumo:
A previously unknown chemical structure, 6-desmethyl-6-ethylerythromycin A (6-ethylErA), was produced through directed genetic manipulation of the erythromycin (Er)-producing organism Saccharopolyspora erythraea. In an attempt to replace the methyl side chain at the C-6 position of the Er polyketide backbone with an ethyl moiety, the methylmalonate-specific acyltransferase (AT) domain of the Er polyketide synthase was replaced with an ethylmalonate-specific AT domain from the polyketide synthase involved in the synthesis of the 16-member macrolide niddamycin. The genetically altered strain was found to produce ErA, however, and not the ethyl-substituted derivative. When the strain was provided with precursors of ethylmalonate, a small quantity of a macrolide with the mass of 6-ethylErA was produced in addition to ErA. Because substrate for the heterologous AT seemed to be limiting, crotonyl-CoA reductase, a primary metabolic enzyme involved in butyryl-CoA production in streptomycetes, was expressed in the strain. The primary macrolide produced by the reengineered strain was 6-ethylErA.
Resumo:
A protein semisynthesis method—expressed protein ligation—is described that involves the chemoselective addition of a peptide to a recombinant protein. This method was used to ligate a phosphotyrosine peptide to the C terminus of the protein tyrosine kinase C-terminal Src kinase (Csk). By intercepting a thioester generated in the recombinant protein with an N-terminal cysteine containing synthetic peptide, near quantitative chemical ligation of the peptide to the protein was achieved. The semisynthetic tail-phosphorylated Csk showed evidence of an intramolecular phosphotyrosine-Src homology 2 interaction and an unexpected increase in catalytic phosphoryl transfer efficiency toward a physiologically relevant substrate compared with the non-tail-phosphorylated control. This work illustrates that expressed protein ligation is a simple and powerful new method in protein engineering to introduce sequences of unnatural amino acids, posttranslational modifications, and biophysical probes into proteins of any size.
Resumo:
In an effort to expand the scope of protein mutagenesis, we have completed the first steps toward a general method to allow the site-specific incorporation of unnatural amino acids into proteins in vivo. Our approach involves the generation of an “orthogonal” suppressor tRNA that is uniquely acylated in Escherichia coli by an engineered aminoacyl-tRNA synthetase with the desired unnatural amino acid. To this end, eight mutations were introduced into tRNA2Gln based on an analysis of the x-ray crystal structure of the glutaminyl-tRNA aminoacyl synthetase (GlnRS)–tRNA2Gln complex and on previous biochemical data. The resulting tRNA satisfies the minimal requirements for the delivery of an unnatural amino acid: it is not acylated by any endogenous E. coli aminoacyl-tRNA synthetase including GlnRS, and it functions efficiently in protein translation. Repeated rounds of DNA shuffling and oligonucleotide-directed mutagenesis followed by genetic selection resulted in mutant GlnRS enzymes that efficiently acylate the engineered tRNA with glutamine in vitro. The mutant GlnRS and engineered tRNA also constitute a functional synthetase–tRNA pair in vivo. The nature of the GlnRS mutations, which occur both at the protein–tRNA interface and at sites further away, is discussed.
Resumo:
Protein–protein interacting surfaces are usually large and intricate, making the rational design of small mimetics of these interfaces a daunting problem. On the basis of a structural similarity between the CDR2-like loop of CD4 and the β-hairpin region of a short scorpion toxin, scyllatoxin, we transferred the side chains of nine residues of CD4, central in the binding to HIV-1 envelope glycoprotein (gp120), to a structurally homologous region of the scorpion toxin scaffold. In competition experiments, the resulting 27-amino acid miniprotein inhibited binding of CD4 to gp120 with a 40 μM IC50. Structural analysis by NMR showed that both the backbone of the chimeric β-hairpin and the introduced side chains adopted conformations similar to those of the parent CD4. Systematic single mutations suggested that most CD4 residues from the CDR2-like loop were reproduced in the miniprotein, including the critical Phe-43. The structural and functional analysis performed suggested five additional mutations that, once incorporated in the miniprotein, increased its affinity for gp120 by 100-fold to an IC50 of 0.1–1.0 μM, depending on viral strains. The resulting mini-CD4 inhibited infection of CD4+ cells by different virus isolates. Thus, core regions of large protein–protein interfaces can be reproduced in miniprotein scaffolds, offering possibilities for the development of inhibitors of protein–protein interactions that may represent useful tools in biology and in drug discovery.
Resumo:
Alcaligenes eutrophus genes encoding the enzymes, β-ketothiolase (phaA), acetoacetyl-CoA reductase (phaB), and polyhydroxyalkanoate synthase (phaC) catalyze the production of aliphatic polyester poly-d-(−)-3-hydroxybutyrate (PHB) from acetyl-CoA. PHB is a thermoplastic polymer that may modify fiber properties when synthesized in cotton. Endogenous β-ketothiolase activity is present in cotton fibers. Hence cotton was transformed with engineered phaB and phaC genes by particle bombardment, and transgenic plants were selected based on marker gene, β-glucuronidase (GUS), expression. Fibers of 10 transgenic plants expressed phaB gene, while eight plants expressed both phaB and phaC genes. Electron microscopy examination of fibers expressing both genes indicated the presence of electron-lucent granules in the cytoplasm. High pressure liquid chromatography, gas chromatography, and mass spectrometry evidence suggested that the new polymer produced in transgenic fibers is PHB. Sixty-six percent of the PHB in fibers is in the molecular mass range of 0.6 × 106 to 1.8 × 106 Da. The presence of PHB granules in transgenic fibers resulted in measurable changes of thermal properties. The fibers exhibited better insulating characteristics. The rate of heat uptake and cooling was slower in transgenic fibers, resulting in higher heat capacity. These data show that metabolic pathway engineering in cotton may enhance fiber properties by incorporating new traits from other genetic sources. This is an important step toward producing new generation fibers for the textile industry.
Resumo:
The construction of cDNA clones encoding large-size RNA molecules of biological interest, like coronavirus genomes, which are among the largest mature RNA molecules known to biology, has been hampered by the instability of those cDNAs in bacteria. Herein, we show that the application of two strategies, cloning of the cDNAs into a bacterial artificial chromosome and nuclear expression of RNAs that are typically produced within the cytoplasm, is useful for the engineering of large RNA molecules. A cDNA encoding an infectious coronavirus RNA genome has been cloned as a bacterial artificial chromosome. The rescued coronavirus conserved all of the genetic markers introduced throughout the sequence and showed a standard mRNA pattern and the antigenic characteristics expected for the synthetic virus. The cDNA was transcribed within the nucleus, and the RNA translocated to the cytoplasm. Interestingly, the recovered virus had essentially the same sequence as the original one, and no splicing was observed. The cDNA was derived from an attenuated isolate that replicates exclusively in the respiratory tract of swine. During the engineering of the infectious cDNA, the spike gene of the virus was replaced by the spike gene of an enteric isolate. The synthetic virus replicated abundantly in the enteric tract and was fully virulent, demonstrating that the tropism and virulence of the recovered coronavirus can be modified. This demonstration opens up the possibility of employing this infectious cDNA as a vector for vaccine development in human, porcine, canine, and feline species susceptible to group 1 coronaviruses.
Resumo:
Substitutions or deletions of domain II loop residues of Bacillus thuringiensis δ-endotoxin CryIAb were constructed using site-directed mutagenesis techniques to investigate their functional roles in receptor binding and toxicity toward gypsy moth (Lymantria dispar). Substitution of loop 2 residue N372 with Ala or Gly (N372A, N372G) increased the toxicity against gypsy moth larvae 8-fold and enhanced binding affinity to gypsy moth midgut brush border membrane vesicles (BBMV) ≈4-fold. Deletion of N372 (D3), however, substantially reduced toxicity (>21 times) as well as binding affinity, suggesting that residue N372 is involved in receptor binding. Interestingly, a triple mutant, DF-1 (N372A, A282G and L283S), has a 36-fold increase in toxicity to gypsy moth neonates compared with wild-type toxin. The enhanced activity of DF-1 was correlated with higher binding affinity (18-fold) and binding site concentrations. Dissociation binding assays suggested that the off-rate of the BBMV-bound mutant toxins was similar to that of the wild type. However, DF-1 toxin bound 4 times more than the wild-type and N372A toxins, and it was directly correlated with binding affinity and potency. Protein blots of gypsy moth BBMV probed with labeled N372A, DF-1, and CryIAb toxins recognized a common 210-kDa protein, indicating that the increased activity of the mutants was not caused by binding to additional receptor(s). The improved binding affinity of N372A and DF-1 suggest that a shorter side chain at these loops may fit the toxin more efficiently to the binding pockets. These results offer an excellent model system for engineering δ-endotoxins with higher potency and wider spectra of target pests by improving receptor binding interactions.