Mechanism of mRNA stability control mediated by AU -rich element: Characterization of cis-elements and trans-factors

Regulation of cytoplasmic deadenylation, the first step in mRNA turnover, has direct impact on the fate of gene expression. AU-rich elements (AREs) found in the 3′ untranslated regions of many labile mRNAs are the most common RNA-destabilizing elements known in mammalian cells. Based on their sequence features and functional properties, AREs can be divided into three classes. Class I or class III ARE directs synchronous deadenylation, whereas class II ARE directs asynchronous deadenylation with the formation of poly(A)-intermediates. Through systematic mutagenesis study, we found that a cluster of five or six copies of AUUUA motifs forming various degrees of reiteration is the key feature dictating the choice between asynchronous versus synchronous deadenylation. A 20–30 nt AU-rich sequence immediately 5 ′ to this cluster of AUUUA motifs can greatly enhance its destabilizing ability and is an integral part of the AREs. These two features are the defining characteristics of class II AREs. ^ To better understand the decay mechanism of AREs, current methods have several limitations. Taking the advantage of tetracycline-regulated promoter, we developed a new transcriptional pulse strategy, Tet-system. By controlling the time and the amount of Tet addition, a pulse of RNA could be generated. Using this new system, we showed that AREs function in both growth- and density-arrested cells. The new strategy offers for the first time an opportunity to investigate control of mRNA deadenylation and decay kinetics in mammalian cells that exhibit physiologically relevant conditions. ^ As a member of heterogeneous nuclear RNA-binding protein, hnRNP D 0/AUF1 displays specific affinities for ARE sequences in vitro . But its in vivo function in ARE-mediated mRNA decay is unclear. AUF1/hnRNP D0 is composed of at least four isoforms derived by alternative RNA splicing. Each isoform exhibits different affinity for ARE sequence in vitro. Here, we examined in vivo effect of AUF1s/hnRNP D0s on degradation of ARE-containing mRNA. Our results showed that all four isoforms exhibit various RNA stabilizing effects in NIH3T3 cells, which are positively correlated with their binding affinities for ARE sequences. Further experiments indicated that AUF1/hnRNP D0 has a general role in modulating the stability of cytoplasmic mRNAs in mammalian cells. ^

A human RNase E-like activity that cleaves RNA sequences involved in mRNA stability control.

We have detected an endoribonucleolytic activity in human cell extracts that processes the Escherichia coli 9S RNA and outer membrane protein A (ompA) mRNA with the same specificity as RNase E from E. coli. The human enzyme was partially purified by ion-exchange chromatography, and the active fractions contained a protein that was detected with antibodies shown to recognize E. coli RNase E. RNA containing four repeats of the destabilizing motif AUUUA and RNA from the 3' untranslated region of human c-myc mRNA were also found to be cleaved by E. coli RNase E and its human counterpart in a fashion that may suggest a role of this activity in mammalian mRNA decay. It was also found that RNA containing more than one AUUUA motif was cleaved more efficiently than RNA with only one or a mutated motif. This finding of a eukaryotic endoribonucleolytic activity corresponding to RNase E indicates an evolutionary conservation of the components of mRNA degradation systems.

Direct support and general support maintenance repair parts and special tools list (including depot maintenance repair parts and special tools) for electronic equipment configurations, Army model AH-1S Helicopter production (less stability-control augmentation system (SCAS)).

"5 March 1985."

Multibody System Mechanics: Modelling, Stability, Control, and Robustness by V. A. Konoplev and A. Cheremensky

BOOK REVIEWS Multibody System Mechanics: Modelling, Stability, Control, and Ro- bustness, by V. A. Konoplev and A. Cheremensky, Mathematics and its Appli- cations Vol. 1, Union of Bulgarian Mathematicians, Sofia, 2001, XXII + 288 pp., $ 65.00, ISBN 954-8880-09-01

REAL TIME FUZZY CONTROLLER FOR QUADROTOR STABILITY CONTROL

In this report, we develop an intelligent adaptive neuro-fuzzy controller by using adaptive neuro fuzzy inference system (ANFIS) techniques. We begin by starting with a standard proportional-derivative (PD) controller and use the PD controller data to train the ANFIS system to develop a fuzzy controller. We then propose and validate a method to implement this control strategy on commercial off-the-shelf (COTS) hardware. An analysis is made into the choice of filters for attitude estimation. These choices are limited by the complexity of the filter and the computing ability and memory constraints of the micro-controller. Simplified Kalman filters are found to be good at estimation of attitude given the above constraints. Using model based design techniques, the models are implemented on an embedded system. This enables the deployment of fuzzy controllers on enthusiast-grade controllers. We evaluate the feasibility of the proposed control strategy in a model-in-the-loop simulation. We then propose a rapid prototyping strategy, allowing us to deploy these control algorithms on a system consisting of a combination of an ARM-based microcontroller and two Arduino-based controllers. We then use a combination of the code generation capabilities within MATLAB/Simulink in combination with multiple open-source projects in order to deploy code to an ARM CortexM4 based controller board. We also evaluate this strategy on an ARM-A8 based board, and a much less powerful Arduino based flight controller. We conclude by proving the feasibility of fuzzy controllers on Commercial-off the shelf (COTS) hardware, we also point out the limitations in the current hardware and make suggestions for hardware that we think would be better suited for memory heavy controllers.

Analysis and Control of Excitation, Field Weakening and Stability in Direct Torque Controlled Electrically Excited Synchronous Motor Drives

Direct torque control (DTC) is a new control method for rotating field electrical machines. DTC controls directly the motor stator flux linkage with the stator voltage, and no stator current controllers are used. With the DTC method very good torque dynamics can be achieved. Until now, DTC has been applied to asynchronous motor drives. The purpose of this work is to analyse the applicability of DTC to electrically excited synchronous motor drives. Compared with asynchronous motor drives, electrically excited synchronous motor drives require an additional control for the rotor field current. The field current control is called excitation control in this study. The dependence of the static and dynamic performance of DTC synchronous motor drives on the excitation control has been analysed and a straightforward excitation control method has been developed and tested. In the field weakening range the stator flux linkage modulus must be reduced in order to keep the electro motive force of the synchronous motor smaller than the stator voltage and in order to maintain a sufficient voltage reserve. The dynamic performance of the DTC synchronous motor drive depends on the stator flux linkage modulus. Another important factor for the dynamic performance in the field weakening range is the excitation control. The field weakening analysis considers both dependencies. A modified excitation control method, which maximises the dynamic performance in the field weakening range, has been developed. In synchronous motor drives the load angle must be kept in a stabile working area in order to avoid loss of synchronism. The traditional vector control methods allow to adjust the load angle of the synchronous motor directly by the stator current control. In the DTC synchronous motor drive the load angle is not a directly controllable variable, but it is formed freely according to the motor’s electromagnetic state and load. The load angle can be limited indirectly by limiting the torque reference. This method is however parameter sensitive and requires a safety margin between the theoretical torque maximum and the actual torque limit. The DTC modulation principle allows however a direct load angle adjustment without any current control. In this work a direct load angle control method has been developed. The method keeps the drive stabile and allows the maximal utilisation of the drive without a safety margin in the torque limitation.

Comparison of BR and QR Eigenvalue Algorithms for Power System Small Signal Stability Analysis

The BR algorithm is a novel and efficient method to find all eigenvalues of upper Hessenberg matrices and has never been applied to eigenanalysis for power system small signal stability. This paper analyzes differences between the BR and the QR algorithms with performance comparison in terms of CPU time based on stopping criteria and storage requirement. The BR algorithm utilizes accelerating strategies to improve its performance when computing eigenvalues of narrowly banded, nearly tridiagonal upper Hessenberg matrices. These strategies significantly reduce the computation time at a reasonable level of precision. Compared with the QR algorithm, the BR algorithm requires fewer iteration steps and less storage space without depriving of appropriate precision in solving eigenvalue problems of large-scale power systems. Numerical examples demonstrate the efficiency of the BR algorithm in pursuing eigenanalysis tasks of 39-, 68-, 115-, 300-, and 600-bus systems. Experiment results suggest that the BR algorithm is a more efficient algorithm for large-scale power system small signal stability eigenanalysis.

Development and validation of a stability-indicating HPLC method for the determination of buclizine hydrochloride in tablets and oral suspension and its application to dissolution studies

A method using liquid chromatography has been developed and validated for determination of buclizine in pharmaceutical formulations and in release studies. Isocratic chromatography was performed on a C18 column with methanol:water (80:20 v/v, pH 2.6) as mobile phase, at a flow rate of 1.0 mL/min, and UV detection at 230 nm. The method was linear, accurate, precise, sensible and robust. The dissolution test was optimized and validated in terms of dissolution medium, apparatus agitation and rotation speed. The presented analytical and dissolution procedures can be conveniently adopted in the quality and stability control of buclizine in tablets and oral suspension.

Stability analysis of power systems described with detailed models by automatic method

The power system stability analysis is approached taking into explicit account the dynamic performance of generators internal voltages and control devices. The proposed method is not a direct method in the usual sense since conclusion for stability or instability is not exclusively based on energy function considerations but it is automatic since the conclusion is achieved without an analyst intervention. The stability test accounts for the nonconservative nature of the system with control devices such as the automatic voltage regulator (AVR) and automatic generation control (AGC) in contrast with the well-known direct methods. An energy function is derived for the system with machines forth-order model, AVR and AGC and it is used to start the analysis procedure and to point out criticalities. The conclusive analysis itself is made by means of a method based on the definition of a region surrounding the equilibrium point where the system net torque is equilibrium restorative. This region is named positive synchronization region (PSR). Since the definition of the PSR boundaries have no dependence on modelling approximation, the PSR test conduces to reliable results. (C) 2008 Elsevier Ltd. All rights reserved.

Control aspects in nonlinear Hill's equation

The Hill's equations-even in the linear original version are a describer of phenomenon having chaotic flavor, giving sometimes very unusual situations. The theory of the so called intervals of instability in the equation provides the precise description for most of these phenomena. Considerations on nonlinearities into the Hill's equation is a quite recent task. The linearized version for almost of these systems it reduces to the Hill's classical linear one. In this paper, some indicative facts are pointed out on the possibility of having the linear system stabilizable and/or exactly controllable. As consequence of such an approach we get results having strong classical aspects, like the one talking about location of parameters in intervals of stability. A result for nonlinear proper periodic controls, is considered too. (C) 2010 Elsevier B.V. All rights reserved.

Control preventivo dinamico de sistemas de energia electrica usando redespacho de generacion y corte de carga

A model for preventive control in electrical systems is presented, taking into account the dynamic aspects of the network. Among these aspects, the effects provoked by perturbations which cause oscillations in synchronous machine angles (transient stability), such as electric equipment outages and short circuits, are presented. The energy function is used to measure the stability of the system using a procedure defined as the security margin. The control actions employed are load shedding and generation reallocation. An application of the methodology to a system located in southern Brazil, which is composed of 10 synchronous machines, 45 busses, and 72 transmission lines. The results confirm the theoretical studies.

Molecular control of vertebrate iron metabolism: mRNA-based regulatory circuits operated by iron, nitric oxide, and oxidative stress.

As an essential nutrient and a potential toxin, iron poses an exquisite regulatory problem in biology and medicine. At the cellular level, the basic molecular framework for the regulation of iron uptake, storage, and utilization has been defined. Two cytoplasmic RNA-binding proteins, iron-regulatory protein-1 (IRP-1) and IRP-2, respond to changes in cellular iron availability and coordinate the expression of mRNAs that harbor IRP-binding sites, iron-responsive elements (IREs). Nitric oxide (NO) and oxidative stress in the form of H2O2 also signal to IRPs and thereby influence cellular iron metabolism. The recent discovery of two IRE-regulated mRNAs encoding enzymes of the mitochondrial citric acid cycle may represent the beginnings of elucidating regulatory coupling between iron and energy metabolism. In addition to providing insights into the regulation of iron metabolism and its connections with other cellular pathways, the IRE/IRP system has emerged as a prime example for the understanding of translational regulation and mRNA stability control. Finally, IRP-1 has highlighted an unexpected role for iron sulfur clusters as post-translational regulatory switches.

Kaasupoljinohjeen toteuttaminen hybridilinja-auton säätöjärjestelmälle

Sähkö- ja hybridiajoneuvot yleistyvät tiukentuvien päästömääräysten seurauksena, koska sähkömoottoritekniikan avulla ajoneuvon kokonaishyötysuhdetta on mahdollista parantaa merkittävästi. Akkujen kapasiteettiin, kokoon, painoon ja latausaikoihin liittyvien ongelmien vuoksi sekä nestemäistä polttoainetta että sähköä hyödyntävä hybriditekniikka on toistaiseksi pelkkään sähkökäyttöön verrattuna usein toimivampi toteutus. Vanhojen hyötyajoneuvojen hybridikonversiot saattavatkin yleistyä, jos voidaan osoittaa, että konversio on toteutettavissa järkevin resurssein ja voidaan laatia konversion teknisiä haasteita helpottavia ohjeita. Syksyllä 2012 käynnistyi Lappeenrannan Teknillisen Yliopiston vetämä CAMBUS-projekti, jossa on tarkoitus kehittää kaupallisesti tarjolla olevia järjestelmiä pätevämpi hybriditekniikka linja-autokäyttöön. Tämä kandidaatintyö pyrkii kirjallisuuskatsauksen keinoin selvittämään, minkälainen kaasupolkimen toteutus olisi tätä hybridikonversiota ajatellen paras kuljettajan kontrolloiman säätöohjeen käytännön toteutukseen. Lähdemateriaalina on käytetty mm. Boschin ja Automobiltechnische Zeitschriftin ajoneuvoalan käsikirjoja ja oppikirjoja, Volkswagenin koulutuskäsikirjoja ja täydentävästi tieteellisiä artikkeleita ja patentteja. Niiden pohjalta on koottu yleisimmät tämänhetkiset tekniset toteutustavat, pohdittu tiedonsiirron ja sähkömagneettisen yhteensopivuuden haasteita, sekä kokonaissäätöjärjestelmän näkökulmasta säätöohjeen suuretta. Selvityksen perusteella polkimen asentotunnistuksen kannalta oleellista on turvallisuus, eli lähinnä mekaaninen kestävyys ja riittävä häiriösuojaus. Ajoneuvossa ilmeneviä voimakkaita magneettikenttiä hyvin sietävä digitaalinen väyläjärjestelmä voi hyvin toteutettuna myös yksinkertaistaa ajoneuvon sähköjärjestelmän muuta toteutusta. Aineistojen perusteella vääntömomentti on luonnollisin valinta ohjaussuureeksi. Vääntömomenttiohjeen avulla voidaan helposti luoda selkeä ja johdonmukainen tuntuma kuljettajalle voimanlähteiden turvalliseen hallintaan ja se helpottaa myös kommunikointia pidonhallintajärjestelmän ja muiden ajoneuvon hallintaan liittyvien järjestelmien kanssa.

Desempenho e estabilidade de parâmetros bioquímicos em materiais de controle líquidos congelados e sólidos liofilizados

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Analyse der Mechanismen zur Regulation der Zellzyklus-abhängigen Stabilität der MARCKS-mRNA

Die Expression des PKC-Hauptsubstrates MARCKS (myristoylated alanine-rich C kinase substrate) wird in Swiss 3T3-Fibroblasten in Abhängigkeit des Zellzyklus durch Variation der mRNA-Stabilität reguliert. In der vorliegenden Arbeit wurde die Beteiligung der 3' nichttranslatierten Region (3'UTR) der MARCKS-mRNA an der Stabilitätskontrolle analysiert. Durch Einsatz der RNase/EMSA-Technik konnten zwei cis-Elemente der MARCKS 3'UTR identifiziert und lokalisiert werden, die mit RNA-bindenden Swiss 3T3-Proteinen (trans-Faktoren) interagieren. Diese neu identifizierten cis-Elemente sind AU-reiche Elemente (ARE) der Klasse III, da sie sehr große Sequenzhomologie zu ARE dieser Klasse aufweisen und der MARCKS 3'UTR, wie für ARE typisch, Instabilität vermitteln.Durch UV-crosslinking wurden vier Proteine mit Molekülmassen von 55, 40, 36 und 30 kDa nachgewiesen, die spezifisch an das 52nt lange Haupt-ARE (MARCKS 52nt) mit unterschiedlicher Affinität binden konnten. Mit Hilfe von rekombinant hergestellten ELAV/Hu-Proteinen und einem ELAV/Hu-spezifischen, affinitätsgereinigten Antiserum konnte eines der vier Proteine (p36) als das ELAV/Hu-Protein HuR identifiziert werden. Die Funktion der ELAV/Hu-Proteine für die Stabilitätskontrolle der MARCKS-mRNA ließ sich durch transiente und stabile Transfektion von HuR und neuronenspezifischem HuD mit dem Tetracyclin induzierbaren Expressionssystem (Tetoff) in Swiss 3T3- bzw. MEF/3T3-Tetoff-Zellen verdeutlichen: Durch Überexpression von HuR und HuD wurde die wachstumsinduzierte Destabilisierung der MARCKS-mRNA bei Wiedereintritt der Zellen in den Zellzyklus unterbunden.