Contribución a la localización en interiores basada en comunicaciones inalámbricas

Determinar con buena precisión la posición en la que se encuentra un terminal móvil, cuando éste se halla inmerso en un entorno de interior (centros comerciales, edificios de oficinas, aeropuertos, estaciones, túneles, etc), es el pilar básico sobre el que se sustentan un gran número de aplicaciones y servicios. Muchos de esos servicios se encuentran ya disponibles en entornos de exterior, aunque los entornos de interior se prestan a otros servicios específicos para ellos. Ese número, sin embargo, podría ser significativamente mayor de lo que actualmente es, si no fuera necesaria una costosa infraestructura para llevar a cabo el posicionamiento con la precisión adecuada a cada uno de los hipotéticos servicios. O, igualmente, si la citada infraestructura pudiera tener otros usos distintos, además del relacionado con el posicionamiento. La usabilidad de la misma infraestructura para otros fines distintos ofrecería la oportunidad de que la misma estuviera ya presente en las diferentes localizaciones, porque ha sido previamente desplegada para esos otros usos; o bien facilitaría su despliegue, porque el coste de esa operación ofreciera un mayor retorno de usabilidad para quien lo realiza. Las tecnologías inalámbricas de comunicaciones basadas en radiofrecuencia, ya en uso para las comunicaciones de voz y datos (móviles, WLAN, etc), cumplen el requisito anteriormente indicado y, por tanto, facilitarían el crecimiento de las aplicaciones y servicios basados en el posicionamiento, en el caso de poderse emplear para ello. Sin embargo, determinar la posición con el nivel de precisión adecuado mediante el uso de estas tecnologías, es un importante reto hoy en día. El presente trabajo pretende aportar avances significativos en este campo. A lo largo del mismo se llevará a cabo, en primer lugar, un estudio de los principales algoritmos y técnicas auxiliares de posicionamiento aplicables en entornos de interior. La revisión se centrará en aquellos que sean aptos tanto para tecnologías móviles de última generación como para entornos WLAN. Con ello, se pretende poner de relieve las ventajas e inconvenientes de cada uno de estos algoritmos, teniendo como motivación final su aplicabilidad tanto al mundo de las redes móviles 3G y 4G (en especial a las femtoceldas y small-cells LTE) como al indicado entorno WLAN; y teniendo siempre presente que el objetivo último es que vayan a ser usados en interiores. La principal conclusión de esa revisión es que las técnicas de triangulación, comúnmente empleadas para realizar la localización en entornos de exterior, se muestran inútiles en los entornos de interior, debido a efectos adversos propios de este tipo de entornos como la pérdida de visión directa o los caminos múltiples en el recorrido de la señal. Los métodos de huella radioeléctrica, más conocidos bajo el término inglés “fingerprinting”, que se basan en la comparación de los valores de potencia de señal que se están recibiendo en el momento de llevar a cabo el posicionamiento por un terminal móvil, frente a los valores registrados en un mapa radio de potencias, elaborado durante una fase inicial de calibración, aparecen como los mejores de entre los posibles para los escenarios de interior. Sin embargo, estos sistemas se ven también afectados por otros problemas, como por ejemplo los importantes trabajos a realizar para ponerlos en marcha, y la variabilidad del canal. Frente a ellos, en el presente trabajo se presentan dos contribuciones originales para mejorar los sistemas basados en los métodos fingerprinting. La primera de esas contribuciones describe un método para determinar, de manera sencilla, las características básicas del sistema a nivel del número de muestras necesarias para crear el mapa radio de la huella radioeléctrica de referencia, junto al número mínimo de emisores de radiofrecuencia que habrá que desplegar; todo ello, a partir de unos requerimientos iniciales relacionados con el error y la precisión buscados en el posicionamiento a realizar, a los que uniremos los datos correspondientes a las dimensiones y realidad física del entorno. De esa forma, se establecen unas pautas iniciales a la hora de dimensionar el sistema, y se combaten los efectos negativos que, sobre el coste o el rendimiento del sistema en su conjunto, son debidos a un despliegue ineficiente de los emisores de radiofrecuencia y de los puntos de captura de su huella. La segunda contribución incrementa la precisión resultante del sistema en tiempo real, gracias a una técnica de recalibración automática del mapa radio de potencias. Esta técnica tiene en cuenta las medidas reportadas continuamente por unos pocos puntos de referencia estáticos, estratégicamente distribuidos en el entorno, para recalcular y actualizar las potencias registradas en el mapa radio. Un beneficio adicional a nivel operativo de la citada técnica, es la prolongación del tiempo de usabilidad fiable del sistema, bajando la frecuencia en la que se requiere volver a capturar el mapa radio de potencias completo. Las mejoras anteriormente citadas serán de aplicación directa en la mejora de los mecanismos de posicionamiento en interiores basados en la infraestructura inalámbrica de comunicaciones de voz y datos. A partir de ahí, esa mejora será extensible y de aplicabilidad sobre los servicios de localización (conocimiento personal del lugar donde uno mismo se encuentra), monitorización (conocimiento por terceros del citado lugar) y seguimiento (monitorización prolongada en el tiempo), ya que todos ellas toman como base un correcto posicionamiento para un adecuado desempeño. ABSTRACT To find the position where a mobile is located with good accuracy, when it is immersed in an indoor environment (shopping centers, office buildings, airports, stations, tunnels, etc.), is the cornerstone on which a large number of applications and services are supported. Many of these services are already available in outdoor environments, although the indoor environments are suitable for other services that are specific for it. That number, however, could be significantly higher than now, if an expensive infrastructure were not required to perform the positioning service with adequate precision, for each one of the hypothetical services. Or, equally, whether that infrastructure may have other different uses beyond the ones associated with positioning. The usability of the same infrastructure for purposes other than positioning could give the opportunity of having it already available in the different locations, because it was previously deployed for these other uses; or facilitate its deployment, because the cost of that operation would offer a higher return on usability for the deployer. Wireless technologies based on radio communications, already in use for voice and data communications (mobile, WLAN, etc), meet the requirement of additional usability and, therefore, could facilitate the growth of applications and services based on positioning, in the case of being able to use it. However, determining the position with the appropriate degree of accuracy using these technologies is a major challenge today. This paper provides significant advances in this field. Along this work, a study about the main algorithms and auxiliar techniques related with indoor positioning will be initially carried out. The review will be focused in those that are suitable to be used with both last generation mobile technologies and WLAN environments. By doing this, it is tried to highlight the advantages and disadvantages of each one of these algorithms, having as final motivation their applicability both in the world of 3G and 4G mobile networks (especially in femtocells and small-cells of LTE) and in the WLAN world; and having always in mind that the final aim is to use it in indoor environments. The main conclusion of that review is that triangulation techniques, commonly used for localization in outdoor environments, are useless in indoor environments due to adverse effects of such environments as loss of sight or multipaths. Triangulation techniques used for external locations are useless due to adverse effects like the lack of line of sight or multipath. Fingerprinting methods, based on the comparison of Received Signal Strength values measured by the mobile phone with a radio map of RSSI Recorded during the calibration phase, arise as the best methods for indoor scenarios. However, these systems are also affected by other problems, for example the important load of tasks to be done to have the system ready to work, and the variability of the channel. In front of them, in this paper we present two original contributions to improve the fingerprinting methods based systems. The first one of these contributions describes a method for find, in a simple way, the basic characteristics of the system at the level of the number of samples needed to create the radio map inside the referenced fingerprint, and also by the minimum number of radio frequency emitters that are needed to be deployed; and both of them coming from some initial requirements for the system related to the error and accuracy in positioning wanted to have, which it will be joined the data corresponding to the dimensions and physical reality of the environment. Thus, some initial guidelines when dimensioning the system will be in place, and the negative effects into the cost or into the performance of the whole system, due to an inefficient deployment of the radio frequency emitters and of the radio map capture points, will be minimized. The second contribution increases the resulting accuracy of the system when working in real time, thanks to a technique of automatic recalibration of the power measurements stored in the radio map. This technique takes into account the continuous measures reported by a few static reference points, strategically distributed in the environment, to recalculate and update the measurements stored into the map radio. An additional benefit at operational level of such technique, is the extension of the reliable time of the system, decreasing the periodicity required to recapture the radio map within full measurements. The above mentioned improvements are directly applicable to improve indoor positioning mechanisms based on voice and data wireless communications infrastructure. From there, that improvement will be also extensible and applicable to location services (personal knowledge of the location where oneself is), monitoring (knowledge by other people of your location) and monitoring (prolonged monitoring over time) as all of them are based in a correct positioning for proper performance.

Centrosome positioning and directionality of cell movements

In several cell types, an intriguing correlation exists between the position of the centrosome and the direction of cell movement: the centrosome is located behind the leading edge, suggesting that it serves as a steering device for directional movement. A logical extension of this suggestion is that a change in the direction of cell movement is preceded by a reorientation, or shift, of the centrosome in the intended direction of movement. We have used a fusion protein of green fluorescent protein (GFP) and γ-tubulin to label the centrosome in migrating amoebae of Dictyostelium discoideum, allowing us to determine the relationship of centrosome positioning and the direction of cell movement with high spatial and temporal resolution in living cells. We find that the extension of a new pseudopod in a migrating cell precedes centrosome repositioning. An average of 12 sec elapses between the initiation of pseudopod extension and reorientation of the centrosome. If no reorientation occurs within approximately 30 sec, the pseudopod is retracted. Thus the centrosome does not direct a cell’s migration. However, its repositioning stabilizes a chosen direction of movement, most probably by means of the microtubule system.

Positioning of two alpha subunit carboxy-terminal domains of RNA polymerase at promoters by two transcription factors

Interactions between the cAMP receptor protein (CRP) and the carboxy-terminal regulatory domain (CTD) of Escherichia coli RNA polymerase α subunit were analyzed at promoters carrying tandem DNA sites for CRP binding using a chemical nuclease covalently attached to α. Each CRP dimer was found to direct the positioning of one of the two α subunit CTDs. Thus, the function of RNA polymerase may be subject to regulation through protein–protein interactions between the two α subunits and two different species of transcription factors.

The importance of reactant positioning in enzyme catalysis: A hybrid quantum mechanics/molecular mechanics study of a haloalkane dehalogenase

Hybrid quantum mechanics/molecular mechanics calculations using Austin Model 1 system-specific parameters were performed to study the SN2 displacement reaction of chloride from 1,2-dichloroethane (DCE) by nucleophilic attack of the carboxylate of acetate in the gas phase and by Asp-124 in the active site of haloalkane dehalogenase from Xanthobacter autotrophicus GJ10. The activation barrier for nucleophilic attack of acetate on DCE depends greatly on the reactants having a geometry resembling that in the enzyme or an optimized gas-phase structure. It was found in the gas-phase calculations that the activation barrier is 9 kcal/mol lower when dihedral constraints are used to restrict the carboxylate nucleophile geometry to that in the enzyme relative to the geometries for the reactants without dihedral constraints. The calculated quantum mechanics/molecular mechanics activation barriers for the enzymatic reaction are 16.2 and 19.4 kcal/mol when the geometry of the reactants is in a near attack conformer from molecular dynamics and in a conformer similar to the crystal structure (DCE is gauche), respectively. This haloalkane dehalogenase lowers the activation barrier for dehalogenation of DCE by 2–4 kcal/mol relative to the single point energies of the enzyme's quantum mechanics atoms in the gas phase. SN2 displacements of this sort in water are infinitely slower than in the gas phase. The modest lowering of the activation barrier by the enzyme relative to the reaction in the gas phase is consistent with mutation experiments.

Synergistic contributions of cyclin-dependant kinase 5/p35 and Reelin/Dab1 to the positioning of cortical neurons in the developing mouse brain

Cyclin-dependent kinase (Cdk) 5 is a unique member of the Cdk family, because Cdk5 kinase activity is detected only in the nervous tissue. Two neuron-specific activating subunits of Cdk5, p35 and p39, have been identified. Overlapping expression pattern of these isoforms in the embryonic mouse brain and the significant residual Cdk5 kinase activity in brain homogenate of the p35−/− mice indicate the redundant functions of the Cdk5 activators in vivo. Severe neuronal migration defects in p35−/−Cdk5 +/− mice further support the idea that the redundant expression of the Cdk5 activators may cause a milder phenotype in p35−/− mice compared with Cdk5−/− mice. Mutant mice lacking either Cdk5 or p35 exhibit certain similarities with Reelin/Dab1-mutant mice in the disorganization of cortical laminar structure in the brain. To elucidate the relationship between Cdk5/p35 and Reelin/Dab1 signaling, we generated mouse lines that have combined defects of these genes. The addition of heterozygosity of either Dab1 or Reelin mutation to p35−/− causes the extensive migration defects of cortical neurons in the cerebellum. In the double-null mice of p35 and either Dab1 or Reelin, additional migration defects occur in the Purkinje cells in the cerebellum and in the pyramidal neurons in the hippocampus. These additional defects in neuronal migration in mice lacking both Cdk5/p35 and Reelin/Dab1 indicate that Cdk5/p35 may contribute synergistically to the positioning of the cortical neurons in the developing mouse brain.

Accuracy of lesion bypass by yeast and human DNA polymerase η

DNA polymerase η (Polη) functions in the error-free bypass of UV-induced DNA lesions, and a defect in Polη in humans causes the cancer-prone syndrome, the variant form of xeroderma pigmentosum. Both yeast and human Polη replicate through a cis-syn thymine-thymine dimer (TT dimer) by inserting two As opposite the two Ts of the dimer. Polη, however, is a low-fidelity enzyme, and it misinserts nucleotides with a frequency of ≈ 10−2 to 10−3 opposite the two Ts of the TT dimer as well as opposite the undamaged template bases. This low fidelity of nucleotide insertion seems to conflict with the role of Polη in the error-free bypass of UV lesions. To resolve this issue, we have examined the ability of human and yeast Polη to extend from paired and mispaired primer termini opposite a TT dimer by using steady-state kinetic assays. We find that Polη extends from mispaired primer termini on damaged and undamaged DNAs with a frequency of ≈ 10−2 to 10−3 relative to paired primer termini. Thus, after the incorporation of an incorrect nucleotide, Polη would dissociate from the DNA rather than extend from the mispair. The resulting primer-terminal mispair then could be subject to proofreading by a 3′→5′ exonuclease. Replication through a TT dimer by Polη then would be more accurate than that predicted from the fidelity of nucleotide incorporation alone.

Interactions between tRNA identity nucleotides and their recognition sites in glutaminyl-tRNA synthetase determine the cognate amino acid affinity of the enzyme.

Sequence-specific interactions between aminoacyl-tRNA synthetases and their cognate tRNAs both ensure accurate RNA recognition and prevent the binding of noncognate substrates. Here we show for Escherichia coli glutaminyl-tRNA synthetase (GlnRS; EC 6.1.1.18) that the accuracy of tRNA recognition also determines the efficiency of cognate amino acid recognition. Steady-state kinetics revealed that interactions between tRNA identity nucleotides and their recognition sites in the enzyme modulate the amino acid affinity of GlnRS. Perturbation of any of the protein-RNA interactions through mutation of either component led to considerable changes in glutamine affinity with the most marked effects seen at the discriminator base, the 10:25 base pair, and the anticodon. Reexamination of the identity set of tRNA(Gln) in the light of these results indicates that its constituents can be differentiated based upon biochemical function and their contribution to the apparent Gibbs' free energy of tRNA binding. Interactions with the acceptor stem act as strong determinants of tRNA specificity, with the discriminator base positioning the 3' end. The 10:25 base pair and U35 are apparently the major binding sites to GlnRS, with G36 contributing both to binding and recognition. Furthermore, we show that E. coli tryptophanyl-tRNA synthetase also displays tRNA-dependent changes in tryptophan affinity when charging a noncognate tRNA. The ability of tRNA to optimize amino acid recognition reveals a novel mechanism for maintaining translational fidelity and also provides a strong basis for the coevolution of tRNAs and their cognate synthetases.

Progress toward chemical accuracy in the computer simulation of condensed phase reactions.

We describe a procedure for the generation of chemically accurate computer-simulation models to study chemical reactions in the condensed phase. The process involves (i) the use of a coupled semiempirical quantum and classical molecular mechanics method to represent solutes and solvent, respectively; (ii) the optimization of semiempirical quantum mechanics (QM) parameters to produce a computationally efficient and chemically accurate QM model; (iii) the calibration of a quantum/classical microsolvation model using ab initio quantum theory; and (iv) the use of statistical mechanical principles and methods to simulate, on massively parallel computers, the thermodynamic properties of chemical reactions in aqueous solution. The utility of this process is demonstrated by the calculation of the enthalpy of reaction in vacuum and free energy change in aqueous solution for a proton transfer involving methanol, methoxide, imidazole, and imidazolium, which are functional groups involved with proton transfers in many biochemical systems. An optimized semiempirical QM model is produced, which results in the calculation of heats of formation of the above chemical species to within 1.0 kcal/mol (1 kcal = 4.18 kJ) of experimental values. The use of the calibrated QM and microsolvation QM/MM (molecular mechanics) models for the simulation of a proton transfer in aqueous solution gives a calculated free energy that is within 1.0 kcal/mol (12.2 calculated vs. 12.8 experimental) of a value estimated from experimental pKa values of the reacting species.

Periodicity of strong nucleosome positioning sites around the chicken adult beta-globin gene may encode regularly spaced chromatin.

Positioned nucleosomes contribute to both the structure and the function of the chromatin fiber and can play a decisive role in controlling gene expression. We have mapped, at high resolution, the translational positions adopted by limiting amounts of core histone octamers reconstituted onto 4.4 kb of DNA comprising the entire chicken adult beta-globin gene, its enhancer, and flanking sequences. The octamer displays extensive variation in its affinity for different positioning sites, the range exhibited being about 2 orders of magnitude greater than that of the initial binding of the octamer. Strong positioning sites are located 5' and 3' of the globin gene and in the second intron but are absent from the coding regions. These sites exhibit a periodicity (approximately 200 bp) similar to the average spacing of nucleosomes on the inactive beta-globin gene in vivo, which could indicate their involvement in packaging the gene into higher-order chromatin structure. Overlapping, alternative octamer positioning sites commonly exhibit spacings of 20 and 40 bp, but not of 10 bp. These short-range periodicities could reflect features of the core particle structure contributing to the pronounced sequence-dependent manner in which the core histone octamer interacts with DNA.