The use of insect cells to identify potent and selective inhibitors of the replication of the Dengue and Chikungunya viruses and unravel their molecular mechanism of action

Dengue and Chikungunya viruses cause the most important arthropod-borne viral infections for humans. These viruses are predominant in tropical and subtropical regions. In addition, these viruses are predominant in tropical and subtropical regions. Dengue mortality rate is around 1.2 to 3.5% and deaths due to chikungunya fever are around 1 in 1000; however, half of chikungunya-infected patients evolve into a chronic state that can persist for months up to years. There are no antiviral drugs available for DENV and CHIKV treatment and prevention. Moreover, vector control strategies have failed so far. Thus, the development of potent inhibitors for a broad spectrum of RNA viruses is urgently needed. We established and characterized a new embryonic insect cell line from Culex quinquefasciatus mosquito. Also we established the flaviviruses and alphavirus replication, both in C6/36 and Lulo insect cell lines, as well as in Vero cell line. In addition we carried out a reference compound library and reference panel of assays and data for DENV, which provides a benchmark for further studies. During this study, a panel of 9 antiviral molecules, with proven in vitro anti-dengue virus activity and that act at different stages of the DENV life cycle, was selected. Finally, Favipiravir or T-705, was identified as inhibitor in vitro and in vivo of alphaviruses and the mutation K291R in nsP4, which is responsible of the polymerase activity, was found as the mode of action in CHIKV. Interestingly, lysine in motif F1 is also highly conserved in positive-stranded RNA viruses and this might explain the broad spectrum of T-705 antiviral activity.

"Functional Upregulation of Ca2+ -Activated K+ Channels in the Development of Substantia Nigra Dopamine Neurons"

Many connections in the basal ganglia are made around birth when animals are exposed to a host of new affective, cognitive, and sensori-motor stimuli. It is thought that dopamine modulates cortico-striatal synapses that result in the strengthening of those connections that lead to desired outcomes. We propose that there must be a time before which stimuli cannot be processed into functional connections, otherwise it would imply an effective link between stimulus, response, and reward in uterus. Consistent with these ideas, we present evidence that early in development dopamine neurons are electrically immature and do not produce high-frequency firing in response to salient stimuli. We ask first, what makes dopamine neurons immature? and second, what are the implications of this immaturity for the basal ganglia? As an answer to the first question, we find that at birth the outward current is small (3nS-V), insensitive to Ca2z, TEA, BK, and SK blockers. Rapidly after birth, the outward current increases to 15nS-V and becomes sensitive to Ca2z, TEA, BK, and SK blockers. We make a detailed analysis of the kinetics of the components of the outward currents and produce a model for BK and SK channels that we use to reproduce the outward current, and to infer the geometrical arrangement of BK and Ca2z channels in clusters. In the first cluster, T-type Ca2z and BK channels are coupled within distances of *20 nm (200 A˚). The second cluster consists of L-type Ca2z and BK channels that are spread over distances of at least 60 nm. As for the second question, we propose that early in development, the mechanism of action selection is in a ‘‘locked-in’’ state that would prevent dopamine neurons from reinforcing cortico-striatal synapses that do not have a functional experiential- based value.

Functional Upregulation of Ca2+ -Activated K+ Channels in the Development of Substantia Nigra Dopamine Neurons

Many connections in the basal ganglia are made around birth when animals are exposed to a host of new affective, cognitive, and sensori-motor stimuli. It is thought that dopamine modulates cortico-striatal synapses that result in the strengthening of those connections that lead to desired outcomes. We propose that there must be a time before which stimuli cannot be processed into functional connections, otherwise it would imply an effective link between stimulus, response, and reward in uterus. Consistent with these ideas, we present evidence that early in development dopamine neurons are electrically immature and do not produce high-frequency firing in response to salient stimuli. We ask first, what makes dopamine neurons immature? and second, what are the implications of this immaturity for the basal ganglia? As an answer to the first question, we find that at birth the outward current is small (3nS-V), insensitive to Ca2+, TEA, BK, and SK blockers. Rapidly after birth, the outward current increases to 15nS-V and becomes sensitive to Ca2+, TEA, BK, and SK blockers. We make a detailed analysis of the kinetics of the components of the outward currents and produce a model for BK and SK channels that we use to reproduce the outward current, and to infer the geometrical arrangement of BK and Ca2+ channels in clusters. In the first cluster, T-type Ca2+ and BK channels are coupled within distances of similar to 20 nm (200 parallel to). The second cluster consists of L-type Ca2+ and BK channels that are spread over distances of at least 60 nm. As for the second question, we propose that early in development, the mechanism of action selection is in a "locked-in" state that would prevent dopamine neurons from reinforcing cortico-striatal synapses that do not have a functional experiential-based value.

The work of psychologists in social policy in Brazil

This study verses on the meeting of psychology with the social policy area. The objective is to discuss how psychologists that work in the Basic Health Units and in the Social Assistance Reference Centers experience their daily practices. Using the interview and participant observation methods, we investigated the modes of action of 13 psychologists involved in these service units. The results indicate that because these contexts are adverse to the traditional forms of action, many professionals experience daily strangeness and discomfort, resulting from their encounters with limiting situations of difficult management. Furthermore, we observed practices and experiences that were marked by feelings of compassion, pity, and resentment. We consider, finally, that the encounter between psychology and social policies requires flexibility in traditional acting modes as well as the pursuit of strategies that consider cultural contexts characteristics, avoiding capture, fatigue and saturation process that stifle the worker.

Development and/or change of the notion of permanent object and operative causality: empirical evidence in the first year of life

This study examines the notion of permanent object during the first year of life, taking into account the controversy of two approaches about the nature of change: developmental change and cognitive change. Using a longitudinal/cross-sectional design, tasks adapted of the subscale of permanent object and operative causality of the Uzgiris-Hunt Scale (Uzgiris and Hunt, 1975) (Uzgiris & Hunt, 1975) were presented to 110 infants of 0, 3, 6 and 9 months-old, which reside in three cities of Colombia. The results showed three types of strategies: (a) Not resolution; (b) Exploratory and (c) Resolution, which follow different trajectories in children’s performance. This allows affirming that adaptive conquests of the cognitive development stay together with the variety of strategies. Using strategies reveals adjustments and transformations of action programs that consolidate the notion of permanent object not necessarily with age, but with self-regulatory processes. Empirical evidence contributes to the understanding of the relations between the emergence of novelty in the development and performance variability

Insurgency in a time of terrorism

Is it possible to separate terrorism from armed insurgency? Are all armed manifestations terrorism? Is terrorism a method of action or a logic for action? This article purports to analyze insurgency in the midst of the anti-terrorist era of the new century. By using Vietnam as an example, the author points out how armed insurgency has as of lately been called “terrorist”, yet stresses that this concept should be carefully and strategically looked through in order to better understand the violent nature of two very different phenomena: transactional and trans-national insurgencies-----¿Se puede escindir el terrorismo de la insurgencia armada? ¿Toda expresión armada es terrorismo? ¿El territorio es un método de acción o una lógica de acción? Este artículo presenta un análisis de la insurgencia a la luz de la era antiterrorista del nuevo siglo. Con la utilización del ejemplo de Vietnam el autor señala cómo la insurgencia armada ha sido señalada de “terrorista”; sin embargo, debe revisarse detenidamente de manera estratégica este concepto para entender mejor la naturaleza de la violencia de dos fenómenos diferentes: la insurgencia transnacional y el terrorismo transnacional.