The physiological significance of β-actin mRNA localization in determining cell polarity and directional motility

β-actin mRNA is localized near the leading edge in several cell types, where actin polymerization is actively promoting forward protrusion. The localization of the β-actin mRNA near the leading edge is facilitated by a short sequence in the 3′ untranslated region, the “zip code.” Localization of the mRNA at this region is important physiologically. Treatment of chicken embryo fibroblasts with antisense oligonucleotides complementary to the localization sequence (zip code) in the 3′ untranslated region leads to delocalization of β-actin mRNA, alteration of cell phenotype, and a decrease in cell motility. To determine the components of this process responsible for the change in cell behavior after β-actin mRNA delocalization, the Dynamic Image Analysis System was used to quantify movement of cells in the presence of sense and antisense oligonucleotides to the zip code. It was found that net path length and average speed of antisense-treated cells were significantly lower than in sense-treated cells. Total path length and the velocity of protrusion of antisense-treated cells were not affected compared with those of control cells. These results suggest that a decrease in persistence of direction of movement and not in velocity results from treatment of cells with zip code-directed antisense oligonucleotides. To test this, direct analysis of directionality was performed on antisense-treated cells and showed a decrease in directionality (net path/total path) and persistence of movement. Less directional movement of antisense-treated cells correlated with a unpolarized and discontinuous distribution of free barbed ends of actin filaments and of β-actin protein. These results indicate that delocalization of β-actin mRNA results in delocalization of nucleation sites and β-actin protein from the leading edge followed by loss of cell polarity and directional movement.

Endogenous Msx1 antisense transcript: In vivo and in vitro evidences, structure, and potential involvement in skeleton development in mammals

Msx1 is a key factor for the development of tooth and craniofacial skeleton and has been proposed to play a pivotal role in terminal cell differentiation. In this paper, we demonstrated the presence of an endogenous Msx1 antisense RNA (Msx1-AS RNA) in mice, rats, and humans. In situ analysis revealed that this RNA is expressed only in differentiated dental and bone cells with an inverse correlation with Msx1 protein. These in vivo data and overexpression of Msx1 sense and AS RNA in an odontoblastic cell line (MO6-G3) showed that the balance between the levels of the two Msx1 RNAs is related to the expression of Msx1 protein. To analyze the impact of this balance in the Msx-Dlx homeoprotein pathway, we analyzed the effect of Msx1, Msx2, and Dlx5 overexpression on proteins involved in skeletal differentiation. We showed that the Msx1-AS RNA is involved in crosstalk between the Msx-Dlx pathways because its expression was abolished by Dlx5. Msx1 was shown to down-regulate a master gene of skeletal cells differentiation, Cbfa1. All these data strongly suggest that the ratio between Msx1 sense and antisense RNAs is a very important factor in the control of skeletal terminal differentiation. Finally, the initiation site for Msx1-AS RNA transcription was located by primer extension in both mouse and human in an identical region, including a consensus TATA box, suggesting an evolutionary conservation of the AS RNA-mediated regulation of Msx1 gene expression.

Molecularly engineered resistance to California serogroup virus replication in mosquito cells and mosquitoes.

Introduction of genetic elements derived from a viral pathogen's genome may be used to reduce the vectorial capacity of mosquitoes for that virus. A double subgenomic Sindbis virus expression system was utilized to transcribe sequences of LaCrosse (LAC) virus small (S) or medium (M) segment RNA in sense or antisense orientation; wild-type Sindbis and LaCrosse viruses have single-stranded RNA genomes, the former being positive sense and the latter being negative sense. Recombinant viruses were generated and used to infect Aedes albopictus (C6/36) mosquito cells, which were challenged with wild-type LAC virus and then assayed for LAC virus replication. Several recombinant viruses containing portions of the LAC S segment were capable of inducing varying degrees of interference to the challenge virus. Cells infected with TE/3'2J/ANTI-S virus, expressing full-length negative-sense S RNA of LAC virus, yielded 3-6 log10TCID50 (tissue culture 50% infective dose) less LAC virus per ml than did cells infected with a double subgenomic sindbis virus containing no LAC insert. When C6/36 cells infected with TE/3'2J/ANTI-S were challenged with closely related heterologous bunyaviruses, a similar inhibitory effect was seen. Adult Ae. triseriatus mosquitoes infected with TE/3'2J/ANTI-S were also resistant to challenge by LAC virus. Organs that were productively infected by the double subgenomic Sindbis virus expressing the LAC anti-S sequences demonstrated little LAC virus or antigen. These studies indicate that expression of carefully selected antiviral sequences derived from the pathogen's genome may result in efficacious molecular viral interference in mosquito cells and, more importantly, in mosquitoes.

The BCL2 major breakpoint region is a sequence- and cell-cycle-specific binding site of the Ku antigen.

The majority of translocations involving BCL2 are very narrowly targeted to three breakpoint clusters evenly spaced over a 100-bp region of the gene's terminal exon. We have recently shown that the immediate upstream boundary of this major breakpoint region (mbr) is a specific recognition site for single-strand DNA (ssDNA) binding proteins on the sense and antisense strands. The downstream flank of the mbr is a helicase binding site. In this report we demonstrate that the helicase and ssDNA binding proteins show reciprocal changes in binding activity over the cell cycle. The helicase is maximally active in G1 and early S phases; the ssDNA binding proteins are maximally active in late S and G2/M phases. An inhibitor of helicase binding appears in late S and G2/M. Finally, at least one component of the helicase binding complex is the Ku antigen. Thus, a protein with helicase activity implicated in repair of double-strand breaks, variable (diversity) joining recombination, and, potentially, cell-cycle regulation is targeted to the BCL2 mbr.

A simple test of the vicarious trial-and-error hypothesis of hippocampal function.

Vicarious trial-and-error (VTE) is a term that Muenzinger and Tolman used to describe the rat's conflict-like behavior before responding to choice. Recently, VTE was proposed as a mechanism alternative to the concept of "cognitive map" in accounts of hippocampal function. That is, many phenomena of impaired learning and memory related to hippocampal interventions may be explained by behavioral first principles: reduced conflicting, incipient, pre-choice tendencies to approach and avoid. The nonspatial black-white discrimination learning and VTE behavior of the rat were investigated. Hippocampal-lesioned and sham-lesioned animals were trained for 25 days (20 trials per day) starting at 60 days of age. Each movement of the head from one discriminative stimulus to the other was counted as a VTE instance. Lesioned rats had fewer VTEs than sham controls, and the former learned much more slowly or never learned. After learning, VTE frequency declined. Male and female rats showed no significant differences in VTE behavior or discrimination learning.

Real time optimization in chemical process: evaluation of strategies, improvements and industrial application.

The increasing economic competition drives the industry to implement tools that improve their processes efficiencies. The process automation is one of these tools, and the Real Time Optimization (RTO) is an automation methodology that considers economic aspects to update the process control in accordance with market prices and disturbances. Basically, RTO uses a steady-state phenomenological model to predict the process behavior, and then, optimizes an economic objective function subject to this model. Although largely implemented in industry, there is not a general agreement about the benefits of implementing RTO due to some limitations discussed in the present work: structural plant/model mismatch, identifiability issues and low frequency of set points update. Some alternative RTO approaches have been proposed in literature to handle the problem of structural plant/model mismatch. However, there is not a sensible comparison evaluating the scope and limitations of these RTO approaches under different aspects. For this reason, the classical two-step method is compared to more recently derivative-based methods (Modifier Adaptation, Integrated System Optimization and Parameter estimation, and Sufficient Conditions of Feasibility and Optimality) using a Monte Carlo methodology. The results of this comparison show that the classical RTO method is consistent, providing a model flexible enough to represent the process topology, a parameter estimation method appropriate to handle measurement noise characteristics and a method to improve the sample information quality. At each iteration, the RTO methodology updates some key parameter of the model, where it is possible to observe identifiability issues caused by lack of measurements and measurement noise, resulting in bad prediction ability. Therefore, four different parameter estimation approaches (Rotational Discrimination, Automatic Selection and Parameter estimation, Reparametrization via Differential Geometry and classical nonlinear Least Square) are evaluated with respect to their prediction accuracy, robustness and speed. The results show that the Rotational Discrimination method is the most suitable to be implemented in a RTO framework, since it requires less a priori information, it is simple to be implemented and avoid the overfitting caused by the Least Square method. The third RTO drawback discussed in the present thesis is the low frequency of set points update, this problem increases the period in which the process operates at suboptimum conditions. An alternative to handle this problem is proposed in this thesis, by integrating the classic RTO and Self-Optimizing control (SOC) using a new Model Predictive Control strategy. The new approach demonstrates that it is possible to reduce the problem of low frequency of set points updates, improving the economic performance. Finally, the practical aspects of the RTO implementation are carried out in an industrial case study, a Vapor Recompression Distillation (VRD) process located in Paulínea refinery from Petrobras. The conclusions of this study suggest that the model parameters are successfully estimated by the Rotational Discrimination method; the RTO is able to improve the process profit in about 3%, equivalent to 2 million dollars per year; and the integration of SOC and RTO may be an interesting control alternative for the VRD process.

Legal, Ethical and Cultural Issues U.S. companies face when moving operations to China.

Beyond free trade agreements governing cross-border commerce, the source of most global economic growth is attributed to business across interconnecting markets. Among the most attractive and complex markets, China stands out. Despite its appeal, American businesses are more likely to fail in China due to an overwhelming desire for an immediate return on investment while neglecting to consider or completely disregarding China's unique legal, ethical and cultural environment. This capstone project will give recommendations to help businesses succeed when entering China and avoid legal, ethical and cultural issues such as the ones that Google, Apple, and Yahoo experienced.

GE models and algorithms for condensed phase equilibrium data regression in ternary systems: limitations and proposals

Phase equilibrium data regression is an unavoidable task necessary to obtain the appropriate values for any model to be used in separation equipment design for chemical process simulation and optimization. The accuracy of this process depends on different factors such as the experimental data quality, the selected model and the calculation algorithm. The present paper summarizes the results and conclusions achieved in our research on the capabilities and limitations of the existing GE models and about strategies that can be included in the correlation algorithms to improve the convergence and avoid inconsistencies. The NRTL model has been selected as a representative local composition model. New capabilities of this model, but also several relevant limitations, have been identified and some examples of the application of a modified NRTL equation have been discussed. Furthermore, a regression algorithm has been developed that allows for the advisable simultaneous regression of all the condensed phase equilibrium regions that are present in ternary systems at constant T and P. It includes specific strategies designed to avoid some of the pitfalls frequently found in commercial regression tools for phase equilibrium calculations. Most of the proposed strategies are based on the geometrical interpretation of the lowest common tangent plane equilibrium criterion, which allows an unambiguous comprehension of the behavior of the mixtures. The paper aims to show all the work as a whole in order to reveal the necessary efforts that must be devoted to overcome the difficulties that still exist in the phase equilibrium data regression problem.

Biogeographical Patterns and Phenological Changes in Lapiedra martinezii Lag. Related to Its Alkaloid Diversity

The aim of this work was to investigate the alkaloid patterns of Lapiedra martinezii and their relation to biogeography and phenology focused in a phylogenetic comparison. Plants from 14 populations of L. martinezii, covering almost its entire distribution area, were subjected to morphological, ecological, and phytochemical analysis. Experiments for different alkaloid-type content are proposed as a new tool for analysis of plant distribution. Several plants were transplanted for weekly observation of their phenological changes, and alkaloids from different plant organs were extracted, listed, and compared. The alkaloid pattern of L. martinezii comprises 49 compounds of homolycorine, lycorine, tazettine, haemantamine, and narciclasine types. The populations located in the north and south margins of the distribution area displayed alkaloid patterns different from those of the central area. Changes in these patterns during their phenological cycle may be related to a better defence for plant reproduction. L. martinezii is an old relict plant, and it has maintained some of the more primitive morphological features and alkaloid profiles of the Mediterranean Amaryllidaceae. The variations in alkaloid content observed could be interpreted in a phylogenetic sense, and those found in their phenological changes, in an adaptive one.

Culture and World Vision: The Cognitive Mythical Mode

Culture is the system of knowledge, from whose meanings the human being screened and selected their understanding of Reality in the broad sense, and interprets and regulates the facts and data of social behavior. In this sense, culture is a program for social action and acting in humans during the process of socialization and social interaction. The meanings of each culture are the cumulative product of collective and individual thinking, in ecological economic, social and political specific situations, so are the expression of each particular cultural historical conjuncture. Moreover, the universal cognitive structure for the apprehension of cultural reality is the World Vision (WV). Due to its importance and significance as substratum of religious and political belief systems, we will gird our study to mythical cognitive mode or mythical WV.

La reapertura de radiotelevisión valenciana. De la manipulación al servicio público

La próxima reapertura de la radiotelevisión valenciana, teniendo en cuenta el fracaso en la gestión de RTVV durante años, suscita interrogantes sobre cómo se está desarrollando el proceso de creación del nuevo ente público. Cuáles son las medidas que se están llevando a cabo para garantizar la independencia política de la nueva corporación y evitar los errores del pasado. El objetivo de este estudio consiste en comparar los mecanismos y herramientas con los que contaba RTVV y con los que se dotará al nuevo ente, ya que a priori marcarán la diferencia entre ambos. En este sentido, la investigación se fundamenta en el análisis de contenido de la legislación tanto de RTVV como de la futura Corporación Valenciana de Medios de Comunicación, teniendo como referencia el informe de expertos Bases per a la renovació de l’espai comunicatiu valencià i la restitució del servei públic de radiotelevisió. Finalmente, se ha podido concluir que algunas herramientas de autorregulación interna propuestas hoy, ya formaban parte de la antigua RTVV. Estos organismos no cumplieron su cometido debido a la politización de los mismos, que en definitiva fue la principal causa de su declive. No obstante, también existieron carencias en cuanto a regulación externa, y que en esta nueva etapa se están incluyendo para aumentar las garantías de independencia y cumplimiento deontológico profesional.

Countermanding in rats as a practical model for investigation of adaptive control of behaviour, lifespan changes in behavioural control and neurotransmitter function

The countermanding paradigm was designed to investigate the ability to cancel a prepotent response when a stop signal is presented and allows estimation of the stop signal response time (SSRT), an otherwise unobservable behaviour. Humans exhibit adaptive control of behaviour in the countermanding task, proactively lengthening response time (RT) in expectation of stopping and reactively lengthening RT following stop trials or errors. Human performance changes throughout the lifespan, with longer RT, SSRT and greater emphasis on post-error slowing reported for older compared to younger adults. Inhibition in the task has generally been improved by drugs that increase extracellular norepinephrine. The current thesis examined a novel choice response countermanding task in rats to explore whether rodent countermanding performance is a suitable model for the study of adaptive control of behaviour, lifespan changes in behavioural control and the role of neurotransmitters in these behaviours. Rats reactively adjusted RT in the countermanding task, shortening RT after consecutive correct go trials and lengthening RT following non-cancelled, but not cancelled stop trials, in sessions with a 10 s, but not a 1 s post-error timeout interval. Rats proactively lengthened RT in countermanding task sessions compared to go trial-only sessions. Together, these findings suggest that rats strategically lengthened RT in the countermanding task to improve accuracy and avoid longer, unrewarded timeout intervals. Next, rats exhibited longer RT and relatively conserved post-error slowing, but no significant change in SSRT when tested at 12, compared to 7 months of age, suggesting that rats exhibit changes in countermanding task performance with aging similar to those observed in humans. Finally, acute administration of yohimbine (1.25, 2.5 mg/kg) and d-amphetamine (0.25, 0.5 mg/kg), which putatively increase extracellular norepinephrine and dopamine respectively, resulted in RT shortening, baseline-dependent effects on SSRT, and attenuated adaptive RT adjustments in rats in the case of d-amphetamine. These findings suggest that dopamine and norepinephrine encouraged motivated, reward-seeking behaviour and supported inhibitory control in an inverted-U-like fashion. Taken together, these observations validate the rat countermanding task for further study of the neural correlates and neurotransmitters mediating adaptive control of behaviour and lifespan changes in behavioural control.

The Mirage of the State: Why the West has failed in Afghanistan, Iraq, Libya, Mali and Central African Republic? Jean Monnet/Robert Schuman Paper Series Vol. 14 No. 15, July 2014

The state still matters. However, the members of the Euro-Atlantic community may be misinterpreting this crucial baseline prior launching their military interventions since 2001. The latest violence and collapse of the state of Iraq after the invasion of Northern Iraq by a radical Sunni Muslim terrorist group, so-called Islamic State of Iraq and Syria (ISIS), demonstrate once again the centrality and requirement of a functioning state in order to maintain violent forces to disrupt domestic and regional stability. Since 2001, the US and its European allies have waged wars against failed-states in order to increase this security and national interests, and then have been involved in some type of state-building.1 This has been the case in Afghanistan, Iraq, Libya, Mali, and Central African Republic (CAR). France went into Mali (2012) and CAR (2013), which preceded two European Union military and civilian Common Security and Defense Policy missions (CSDP), in order to avoid the collapse of these two states. The threat of the collapse of both states was a concern for the members of the Euro-Atlantic community as it could have spread to the region and causing even greater instabilities. In Mali, the country was under radical Islamic pressures coming from the North after the collapse of Libya ensuing the 2011 Western intervention, while in CAR it was mainly an ethno-religious crisis. Failed states are a real concern, as they can rapidly become training grounds for radical groups and permitting all types of smuggling and trafficking.2 In Mali, France wanted to protect its large French population and avoid the fall of Mali in the hands of radical Islamic groups directly or indirectly linked to Al-Qaeda. A fallen Mali could have destabilized the region of the Sahel and ultimately affected the stability of Southern European borders. France wanted to avoid the development of a safe haven across the Sahel where movements of people and goods are uncontrolled and illegal.3 Since the end of the Cold War, Western powers have been involved in stabilizing neighborhoods and regions, like the Balkans, Africa, and Middle East, which at the exceptions of the Balkans, have led to failed policies. 9/11 changes everything. The US, under President George W. Bush, started to wage war against terrorism and all states link to it. This started a period of continuous Western interventions in this post-9/11 era in Afghanistan, Iraq, Libya, Mali and CAR. If history has demonstrated one thing, the members of the Euro-Atlantic community are struggling and will continue to struggle to stabilize Afghanistan, Iraq, Libya, Mali and Central African Republic (CAR) for one simple reason: no clear endgame. Is it the creation of a state à la Westphalian in order to permit these states to operate as the sole guarantor of security? Or is the reestablishment of status quo in these countries permitting to exit and end Western operations? This article seeks to analyze Western interventions in these five countries in order to reflect on the concept of the state and the erroneous starting point for each intervention.4 In the first part, the political status of each country is analyzed in order to understand the internal and regional crisis. In a second time, the concept of the state, framed into the Buzanian trinity, is discussed and applied to the cases. In the last part the European and American civilian-military doctrines are examined in accordance with their latest military interventions and in their broader spectrum.

The Ukraine crisis and the demise of the European Security order. EPC Policy Brief, 1 December 2015

The annexation of Crimea by Russia and the subsequent war in the Donbas have delivered a severe blow to the European security order, and have increased tensions between the West and Russia to a level unprecedented since the end of the Cold War. In this context, it would be difficult to start negotiating a new commitment to the principles of the European security order. In this Policy Brief, Paul Ivan analyses the Ukraine crisis and its effect on European security and calls for the EU and NATO to stand firm on their own principles, but also to engage carefully with Russia to develop mechanisms to contain risks and avoid escalation and military confrontation, whether in Syria, the Baltic, the Mediterranean or the Black seas. The recent shooting down of a Russian jet by Turkish forces makes clear the need to prevent such incidents with potential rapid escalatory dynamics.

The novels and letters of Jane Austen;

v. 1-2. Sense and sensibility.--v. 3-4. Pride and prejudice.--v. 5-6. Mansfield Park.--v. 7-8. Emma.--v. 9. Northanger Abbey.--v. 10. Persuasion.--v. 11. Lady Susan. The Watsons. Letters.--v. 12. Letters.