Growth cone MKK7 mRNA targeting regulates MAP1b-dependent microtubule bundling to control neurite elongation

Local mRNA translation in neurons has been mostly studied during axon guidance and synapse formation but not during initial neurite outgrowth. We performed a genome-wide screen for neurite-enriched mRNAs and identified an mRNA that encodes mitogen-activated protein kinase kinase 7 (MKK7), a MAP kinase kinase (MAPKK) for Jun kinase (JNK). We show that MKK7 mRNA localizes to the growth cone where it has the potential to be translated. MKK7 is then specifically phosphorylated in the neurite shaft, where it is part of a MAP kinase signaling module consisting of dual leucine zipper kinase (DLK), MKK7, and JNK1. This triggers Map1b phosphorylation to regulate microtubule bundling leading to neurite elongation. We propose a model in which MKK7 mRNA localization and translation in the growth cone allows for a mechanism to position JNK signaling in the neurite shaft and to specifically link it to regulation of microtubule bundling. At the same time, this uncouples activated JNK from its functions relevant to nuclear translocation and transcriptional activation.

The role of Lmx1b in the control of dorsoventral compartmentalization in the distal mesenchyme of the developing mouse limb

Lmx1b encodes a LIM-homeodomain transcription factor required for dorso-ventral (D-V) patterning in the mesenchyme of the vertebrate limb. In the absence of Lmx1b function, limbs exhibit a bi-ventral pattern indicating that Lmx1b is required for cells to adopt a dorsal cell fate. However, how Lmx1b specifies dorsal cell fates in the mesenchyme of the distal limb is unknown. Lmx1b is initially expressed throughout the dorsal and ventral limb bud mesenchyme, then becomes dorsally restricted around E10.5. At this stage, there is a sharp boundary between Lmx1b expressing and Lmx1b non-expressing cells. How the dorso-ventral Lmx1b expression boundary is formed and maintained is currently unknown. One mechanism that may contribute to establishing and/or maintaining the Lmx1b expression boundary is if the dorsal mesenchyme is a lineage-based compartment, where different groups of non-mingling cells are separated. Compartment formation has been proposed to rely on compartment-specific selector gene activity which functions to restrict cells to a compartment and specifies the identity of cells within that compartment. Based on the evidence that the dorsal limb identity relies on the expression of Lmx1b in the dorsal half of the limb mesenchyme, we hypothesized that Lmx1b might function as a dorsal limb bud mesenchyme selector gene to set up a dorsal compartment. To test this hypothesis, we developed an inducible CreERT2/ loxP based fate mapping approach that permanently marks Lmx1b wild-type and mutant cells and examined the distribution of their descendents in the developing limb. Our data is the first to show that dorso-ventral lineage compartments exist in the limb bud mesenchyme. Furthermore, Lmx1b is required for maintenance of the dorso-ventral compartment lineage boundary. The behavior of Lmx1b mutant cells that cross into the ventral mesenchyme, as well as previous chimera analysis in which mutant cells spread evenly in the ventral limb and form patches in the dorsal side, suggest that cell affinity differences prevent intermingling of dorsal and ventral cells. ^

GENOME-WIDE PROFILING UNVEILS CRITICIAL FUNCTIONS OF p53 IN HUMAN EMBRYONIC STEM CELLS

Embryonic stem cells (ESCs) possess two unique characteristics: infinite self-renewal and the potential to differentiate into almost every cell type (pluripotency). Recently, global expression analyses of metastatic breast and lung cancers revealed an ESC-like expression program or signature, specifically for cancers that are mutant for p53 function. Surprisingly, although p53 is widely recognized as the guardian of the genome, due to its roles in cell cycle checkpoints, programmed cell death or senescence, relatively little is known about p53 functions in normal cells, especially in ESCs. My hypothesis is that p53 has specific transcription regulatory functions in human ESCs (hESCs) that a) oppose pluripotency and b) protect the stem cell genome in response to DNA damage and stress signaling. In mouse ESCs, these roles are believed to coincide, as p53 promotes differentiation in response to DNA damage, but this is unexplored in hESCs. To determine the biological roles of p53, specifically in hESCs, we mapped genome-wide chromatin interactions of p53 by chromatin immunoprecipitation and massively parallel tag sequencing (ChIP-Seq), and did so under three VIdifferent conditions of hESC status: pluripotency, differentiation-initiated and DNA-damage-induced. ChIP-Seq showed that p53 is enriched at distinct, induction-specific gene loci during each of these different conditions. Microarray gene expression analysis and functional annotation of the distinct p53-target genes revealed that p53 regulates specific genes encoding developmental regulators, which are expressed in differentiation-initiated but not DNA- damaged hESCs. We further discovered that, in response to differentiation signaling, p53 binds regions of chromatin that are repressed but also poised for rapid activation by core pluripotency factors OCT4 and NANOG in pluripotent hESCs. In response to DNA damage, genes associated with migration and motility are targeted by p53; whereas, the prime targets of p53 in control of cell death are conserved for p53 regulation in both differentiation and DNA damage. Our genome-wide profiling and bioinformatics analyses show that p53 occupies a special set of developmental regulatory genes during early differentiation of hESCs and functions in an induction-specific manner. In conclusion, our research unveiled previously unknown functions of p53 in ESC biology, which augments our understanding of one of the most deregulated proteins in human cancers.

{\it cis\/} -acting elements and {\it trans\/} -acting factors that control serum amyloid A gene expression during inflammation

To understand how the serum amyloid A (SAA) genes are regulated, the cis-acting elements and trans-acting factors involved in the regulation of mouse SAA3 and rat SAA1 genes expression during inflammation were analyzed.^ To identify DNA sequences involved in the liver-specific expression of the mouse SAA3 gene, the 5$\sp\prime$ flanking region of this gene was analyzed by transient transfection studies. Results suggest that C/EBP, a liver-enriched transcription factor, plays an important role for the enhanced expression of the mouse SAA3 gene in hepatocytes.^ Transfection studies of the regulation of the expression of rat SAA1 gene indicated that a 322 bp fragment ($-$304 to +18) of the gene contains sufficient information for cytokine-induced expression of the reporter gene in a liver cell-specific manner. Further functional analysis of the 5$\sp\prime$ flanking region of the rat SAA1 gene demonstrated that a 65 bp DNA fragment ($-$138/$-$73) can confer cytokine-inducibility onto a heterologous promoter both in liver and nonliver cells. DNase I footprint and gel retardation assays identified five putative cis-regulatory elements within the 5$\sp\prime$ flanking region of the gene: one inducible element, a NF$\kappa$B binding site and four constitutive elements. Two constitutive elements, footprint regions I and III, were identified as C/EBP binding sites with region III having over a 10-fold higher affinity for C/EBP binding than region I. Functional analysis of the cis-elements indicated that C/EBP(I) and C/EBP(III) confer liver cell-specific activation onto a heterologous promoter, while sequences corresponding to the NF$\kappa$B element and C/EBP(I) impart cytokine responsiveness onto the heterologous promoter. These results suggest that C/EBP(I) possesses two functions: liver-specific activation and cytokine responsiveness. The identification of two cytokine responsive elements (NF$\kappa$B and C/EBP(I)), and two liver-specific elements (C/EBP(I) and C/EBP(III)) implies that multiple cis-acting elements are involved in the regulation of the expression of the rat SAA1 gene. The tissue-specific and cytokine-induced expression of rat SAA1 gene is likely the result of the interactions of these cis-acting elements with their cognate trans-acting factors as well as the interplay between the different cis-acting elements and their binding factors. (Abstract shortened with permission of author.) ^

Control of {\it recA\/} expression in {\it Escherichia coli}

The recA gene is essential for SOS response induction, for inducible DNA repair and for homologous recombination in E. coli. The level of recA expression is significant for these functions. A basal level of about 1000 molecules of RecA protein is sufficient for homologous recombination of the cell and is essential for the induction of the SOS response. Based on previous observations, two models regarding the origin of the basal RecA protein were postulated. One was that it comes from the leaky expression of the LexA repressed promoter. The other was that it is from another weak but constitutive promoter. The first part of this thesis is to study these possibilities. An $\Omega$ cartridge containing the transcription terminator of gene 32 of T4 phage was exploited to define a second promoter for recA expression. Insertion of this $\Omega$ cartridge downstream of the known promoter gave rise to only minor expression. Purification and N-terminus sequencing of the RecA protein from the insertion mutant did not support the existence of a second promoter. To determine whether the basal RecA is due to the leaky expression of the known LexA repressed promoter, recA expression of a SOS induction minus strain (basal level expression of recA) was compared with that of a recA promoter down mutation recA1270. The result demonstrated that there is leaky expression from the LexA repressed promoter. All the evidence supports the conclusion that there is only one promoter for both basal and induced expression levels of recA.^ Several translation enhancer sequences which are complementary to different regions of the 16S rRNA were found to exist in recA mRNA. The leader sequence of recA mRNA is highly complementary to a region of the 16S rRNA. Thus it appeared that recA expression could be regulated at post-transcriptional levels. The second part of this thesis is focused on the study of the post-transcriptional control of recA expression. Deletions of the complementary regions were created to examine their effect on recA expression. The results indicated that all of the complementary regions were important for the normal expression of recA and their effects were post-transcriptional. RNA secondary structures of wild type recA mRNA was inspected and a stem-loop structure was revealed. The expression down mutations at codon 10 and 11 were found to stabilize this structure. The conclusions of the second part of this thesis are that there is post-transcriptional control for recA expression and the leader sequence of recA mRNA plays more than one role in the control of recA expression. ^

Adaptive fuzzy knowledge-based systems for control metabots' mobility on virtual environments

The confluence of three-dimensional (3D) virtual worlds with social networks imposes on software agents, in addition to conversational functions, the same behaviours as those common to human-driven avatars. In this paper, we explore the possibilities of the use of metabots (metaverse robots) with motion capabilities in complex virtual 3D worlds and we put forward a learning model based on the techniques used in evolutionary computation for optimizing the fuzzy controllers which will subsequently be used by metabots for moving around a virtual environment.

New methods for the estimation of Takagi-Sugeno model based extended Kalman filter and its applications to optimal control for nonlinear systems

This paper describes new approaches to improve the local and global approximation (matching) and modeling capability of Takagi–Sugeno (T-S) fuzzy model. The main aim is obtaining high function approximation accuracy and fast convergence. The main problem encountered is that T-S identification method cannot be applied when the membership functions are overlapped by pairs. This restricts the application of the T-S method because this type of membership function has been widely used during the last 2 decades in the stability, controller design of fuzzy systems and is popular in industrial control applications. The approach developed here can be considered as a generalized version of T-S identification method with optimized performance in approximating nonlinear functions. We propose a noniterative method through weighting of parameters approach and an iterative algorithm by applying the extended Kalman filter, based on the same idea of parameters’ weighting. We show that the Kalman filter is an effective tool in the identification of T-S fuzzy model. A fuzzy controller based linear quadratic regulator is proposed in order to show the effectiveness of the estimation method developed here in control applications. An illustrative example of an inverted pendulum is chosen to evaluate the robustness and remarkable performance of the proposed method locally and globally in comparison with the original T-S model. Simulation results indicate the potential, simplicity, and generality of the algorithm. An illustrative example is chosen to evaluate the robustness. In this paper, we prove that these algorithms converge very fast, thereby making them very practical to use.

Grid-based histogram arithmetic for the probabilistic analysis of functions

The selection of predefined analytic grids (partitions of the numeric ranges) to represent input and output functions as histograms has been proposed as a mechanism of approximation in order to control the tradeoff between accuracy and computation times in several áreas ranging from simulation to constraint solving. In particular, the application of interval methods for probabilistic function characterization has been shown to have advantages over other methods based on the simulation of random samples. However, standard interval arithmetic has always been used for the computation steps. In this paper, we introduce an alternative approximate arithmetic aimed at controlling the cost of the interval operations. Its distinctive feature is that grids are taken into account by the operators. We apply the technique in the context of probability density functions in order to improve the accuracy of the probability estimates. Results show that this approach has advantages over existing approaches in some particular situations, although computation times tend to increase significantly when analyzing large functions.

Advanced predictive quality control strategy involving different facilities

There are many industries that use highly technological solutions to improve quality in all of their products. The steel industry is one example. Several automatic surface-inspection systems are used in the steel industry to identify various types of defects and to help operators decide whether to accept, reroute, or downgrade the material, subject to the assessment process. This paper focuses on promoting a strategy that considers all defects in an integrated fashion. It does this by managing the uncertainty about the exact position of a defect due to different process conditions by means of Gaussian additive influence functions. The relevance of the approach is in making possible consistency and reliability between surface inspection systems. The results obtained are an increase in confidence in the automatic inspection system and an ability to introduce improved prediction and advanced routing models. The prediction is provided to technical operators to help them in their decision-making process. It shows the increase in improvement gained by reducing the 40 % of coils that are downgraded at the hot strip mill because of specific defects. In addition, this technology facilitates an increase of 50 % in the accuracy of the estimate of defect survival after the cleaning facility in comparison to the former approach. The proposed technology is implemented by means of software-based, multi-agent solutions. It makes possible the independent treatment of information, presentation, quality analysis, and other relevant functions.

A new approach to fuzzy estimation of Takagi-Sugeno model and its applications to optimal control for nonlinear systems

An efficient approach is presented to improve the local and global approximation and modelling capability of Takagi-Sugeno (T-S) fuzzy model. The main aim is obtaining high function approximation accuracy. The main problem is that T-S identification method cannot be applied when the membership functions are overlapped by pairs. This restricts the use of the T-S method because this type of membership function has been widely used during the last two decades in the stability, controller design and are popular in industrial control applications. The approach developed here can be considered as a generalized version of T-S method with optimized performance in approximating nonlinear functions. A simple approach with few computational effort, based on the well known parameters' weighting method is suggested for tuning T-S parameters to improve the choice of the performance index and minimize it. A global fuzzy controller (FC) based Linear Quadratic Regulator (LQR) is proposed in order to show the effectiveness of the estimation method developed here in control applications. Illustrative examples of an inverted pendulum and Van der Pol system are chosen to evaluate the robustness and remarkable performance of the proposed method and the high accuracy obtained in approximating nonlinear and unstable systems locally and globally in comparison with the original T-S model. Simulation results indicate the potential, simplicity and generality of the algorithm.

Herramienta de control de dispositivo móvil mediante un ordenador

Este trabajo fin de máster se integra con el sistema Localiza, un sistema previamente desarrollado para la localización bajo demanda de personas que requieran un cierto grado de supervisión. El proyecto amplia las funciones del sistema Localiza, añadiendo una nueva funcionalidad que permita a un usuario con movilidad reducida controlar su dispositivo móvil a través de su ordenador personal. Este proyecto se integra con el proyecto Localiza bajo el título: “Desarrollo de una herramienta software para el manejo de un teléfono móvil adaptada a personas con discapacidad física severa”. El proyecto citado se centra en el desarrollo de una aplicación móvil, que se comunicara con el ordenador personal del usuario. El desarrollo del sistema residente en el ordenador personal, es el ámbito central que ocupa a este trabajo fin de Master. El usuario final al que está destinada la aplicación desarrollada en este proyecto, es un usuario con grado de discapacidad motórica, de forma que con ligeros movimientos de cabeza sea capaz de controlar remotamente el terminal móvil a través de un ordenador personal. El objetivo principal del proyecto es el control remoto de un terminal móvil desde un ordenador personal. La comunicación entre el terminal móvil y el ordenador personal se ha realizado bajo el protocolo Bluetooth. Para desarrollar la aplicación residente en el ordenador personal, se ha utilizado la plataforma Java. SUMMARY. This Master Tesis develops an application, which is intended to provide an added value to the already existing project Localiza, on-demand position system for people with severe disabilities. This project extends the functions of the Localiza system, adding a new feature that allows a user with limited mobility to control their mobile device. This project is integrated with the project under the title: “Desarrollo de una herramienta software para el manejo de un teléfono móvil adaptada a personas con discapacidad física severa”. The above project it is focused on the development of mobile application and the development of the application that resides in the personal computer is the main work of this project. Both projects are closely related and together they complement. The end-user of the application that is developed in this project is a person with motor disabilities. This person may control the computer mouse with slight head movements. The aim of this project is to facilitate the access to the personal computer and to the mobile telephony environment for disabled people. The Communication between the mobile and personal computer has been conducted under the Bluetooth protocol. To develop the application resident on the personal computer have been used the Java platform.

Variable structure control with chattering elimination and guaranteed stability for a generalized T-S model

In this paper, a fuzzy based Variable Structure Control (VSC) with guaranteed stability is presented. The main objective is to obtain an improved performance of highly non-linear unstable systems. The main contribution of this work is that, firstly, new functions for chattering reduction and error convergence without sacrificing invariant properties are proposed, which is considered the main drawback of the VSC control. Secondly, the global stability of the controlled system is guaranteed.The well known weighting parameters approach, is used in this paper to optimize local and global approximation and modeling capability of T-S fuzzy model.A one link robot is chosen as a nonlinear unstable system to evaluate the robustness, effectiveness and remarkable performance of optimization approach and the high accuracy obtained in approximating nonlinear systems in comparison with the original T-S model. Simulation results indicate the potential and generality of the algorithm. The application of the proposed FLC-VSC shows that both alleviation of chattering and robust performance are achieved with the proposed FLC-VSC controller. The effectiveness of the proposed controller is proven in front of disturbances and noise effects.

Variable structure control with chattering reduction of a generalized T-S model

In this paper, a fuzzy logic controller (FLC) based variable structure control (VSC) is presented. The main objective is to obtain an improved performance of highly non-linear unstable systems. New functions for chattering reduction and error convergence without sacrificing invariant properties are proposed. The main feature of the proposed method is that the switching function is added as an additional fuzzy variable and will be introduced in the premise part of the fuzzy rules; together with the state variables. In this work, a tuning of the well known weighting parameters approach is proposed to optimize local and global approximation and modelling capability of the Takagi-Sugeno (T-S) fuzzy model to improve the choice of the performance index and minimize it. The main problem encountered is that the T-S identification method can not be applied when the membership functions are overlapped by pairs. This in turn restricts the application of the T-S method because this type of membership function has been widely used in control applications. The approach developed here can be considered as a generalized version of the T-S method. An inverted pendulum mounted on a cart is chosen to evaluate the robustness, effectiveness, accuracy and remarkable performance of the proposed estimation approach in comparison with the original T-S model. Simulation results indicate the potential, simplicity and generality of the estimation method and the robustness of the chattering reduction algorithm. In this paper, we prove that the proposed estimation algorithm converge the very fast, thereby making it very practical to use. The application of the proposed FLC-VSC shows that both alleviation of chattering and robust performance are achieved.

Effect of control method on impedance-based interactions in a buck converter

All the interconnected regulated systems are prone to impedance-based interactions making them sensitive to instability and transient-performance degradation. The applied control method affects significantly the characteristics of the converter in terms of sensitivity to different impedance interactions. This paper provides for the first time the whole set of impedance-type internal parameters and the formulas according to which the interaction sensitivity can be fully explained and analyzed. The formulation given in this paper can be utilized equally either based on measured frequency responses or on predicted analytic transfer functions. Usually, the distributed dc-dc systems are constructed by using ready-made power modules without having thorough knowledge on the actual power-stage and control-system designs. As a consequence, the interaction characterization has to be based on the frequency responses measureable via the input and output terminals. A buck converter with four different control methods is experimentally characterized in frequency domain to demonstrate the effect of control method on the interaction sensitivity. The presented analytical models are used to explain the phenomena behind the changes in the interaction sensitivity.

A novel topology control approach to maintain the node degree in dynamic wireless sensor networks

Topology control is an important technique to improve the connectivity and the reliability of Wireless Sensor Networks (WSNs) by means of adjusting the communication range of wireless sensor nodes. In this paper, a novel Fuzzy-logic Topology Control (FTC) is proposed to achieve any desired average node degree by adaptively changing communication range, thus improving the network connectivity, which is the main target of FTC. FTC is a fully localized control algorithm, and does not rely on location information of neighbors. Instead of designing membership functions and if-then rules for fuzzy-logic controller, FTC is constructed from the training data set to facilitate the design process. FTC is proved to be accurate, stable and has short settling time. In order to compare it with other representative localized algorithms (NONE, FLSS, k-Neighbor and LTRT), FTC is evaluated through extensive simulations. The simulation results show that: firstly, similar to k-Neighbor algorithm, FTC is the best to achieve the desired average node degree as node density varies; secondly, FTC is comparable to FLSS and k-Neighbor in terms of energy-efficiency, but is better than LTRT and NONE; thirdly, FTC has the lowest average maximum communication range than other algorithms, which indicates that the most energy-consuming node in the network consumes the lowest power.