Marcelo Cohen : Las fundaciones de la ciencia ficción (parte II: La comunidad)

El presente trabajo constituye una continuación de "Marcelo Cohen. Las fundaciones de la ciencia ficción", texto publicado en esta misma revista en el número 19 del año 2013. Allí ya se anticipaba que el problema de la formulación ontológica subjetiva en Cohen suponía un interrogante en torno a la comunidad. En esa instancia, se dejaron asimismo esbozados algunos aspectos del problema del estar-en-común para ser desarrollados en futuros trabajos. En sintonía con dicha propuesta, se retoman aquí los problemas de la ontología coheniana para luego postular que el problema de la comunidad ya se encuentra en germen en el mismo paradigma que el autor recupera y reformula. En Cohen, la propuesta romántica de una unión entre arte-ciencia-filosofía-religión-política se reactualiza. Aquí se analizan sus inflexiones.

CRÍTICOS & CRITICADOS A CRÍTICA CINEMATOGRÁFICA E SUA RELAÇÃO COM OS CINEASTAS DA RETOMADA

O objetivo deste estudo é analisar a crítica cinematográfica durante o período chamado de Retomada do cinema nacional. Foram escolhidos os maiores veículos de circulação da região Sudeste do País, os jornais O Globo (Rio de Janeiro), Folha de S.Paulo e O Estado de S.Paulo (São Paulo) e a revista Veja para fazer uma análise de conteúdo das críticas das seis maiores bilheterias da Retomada para, em seguida, conferir a recepção das mesmas pelos diretores desses filmes, por meio de entrevistas semi-estruturadas. A intenção é analisar quais conflitos permeiam a relação entre os críticos de cinema e os cineastas, a fim de contribuir para o melhor entendimento do trabalho de dois elementos dos mais importantes da área de cinema. O confronto do material da análise com as entrevistas confirmou a hipótese da existência de conflitos de valores e opiniões entre os dois lados e permitiu identificar pré-julgamentos, simpatias e antipatias, análises emotivas e não-fundamentadas de alguns críticos e cineastas, mas também opiniões e valores fundamentados de outros, demonstrando uma rica diversidade que não se encaixa em uma única definição.(AU)

CRÍTICOS & CRITICADOS A CRÍTICA CINEMATOGRÁFICA E SUA RELAÇÃO COM OS CINEASTAS DA RETOMADA

O objetivo deste estudo é analisar a crítica cinematográfica durante o período chamado de Retomada do cinema nacional. Foram escolhidos os maiores veículos de circulação da região Sudeste do País, os jornais O Globo (Rio de Janeiro), Folha de S.Paulo e O Estado de S.Paulo (São Paulo) e a revista Veja para fazer uma análise de conteúdo das críticas das seis maiores bilheterias da Retomada para, em seguida, conferir a recepção das mesmas pelos diretores desses filmes, por meio de entrevistas semi-estruturadas. A intenção é analisar quais conflitos permeiam a relação entre os críticos de cinema e os cineastas, a fim de contribuir para o melhor entendimento do trabalho de dois elementos dos mais importantes da área de cinema. O confronto do material da análise com as entrevistas confirmou a hipótese da existência de conflitos de valores e opiniões entre os dois lados e permitiu identificar pré-julgamentos, simpatias e antipatias, análises emotivas e não-fundamentadas de alguns críticos e cineastas, mas também opiniões e valores fundamentados de outros, demonstrando uma rica diversidade que não se encaixa em uma única definição.(AU)

Effects of mutations involving cell division, recombination, and chromosome dimer resolution on a priA2∷kan mutant

Recombinational repair of replication forks can occur either to a crossover (XO) or noncrossover (non-XO) depending on Holliday junction resolution. Once the fork is repaired by recombination, PriA is important for restarting these forks in Escherichia coli. PriA mutants are Rec− and UV sensitive and have poor viability and 10-fold elevated basal levels of SOS expression. PriA sulB mutant cells and their nucleoids were studied by differential interference contrast and fluorescence microscopy of 4′,6-diamidino-2-phenylindole-stained log phase cells. Two populations of cells were seen. Eighty four percent appeared like wild type, and 16% of the cells were filamented and had poorly partitioned chromosomes (Par−). To probe potential mechanisms leading to the two populations of cells, mutations were added to the priA sulB mutant. Mutating sulA or introducing lexA3 decreased, but did not eliminate filamentation or defects in partitioning. Mutating either recA or recB virtually eliminated the Par− phenotype. Filamentation in the recB mutant decreased to 3%, but increased to 28% in the recA mutant. The ability to resolve and/or branch migrate Holliday junctions also appeared crucial in the priA mutant because removing either recG or ruvC was lethal. Lastly, it was tested whether the ability to resolve chromosome dimers caused by XOs was important in a priA mutant by mutating dif and the C-terminal portion of ftsK. Mutation of dif showed no change in phenotype whereas ftsK1∷cat was lethal with priA2∷kan. A model is proposed where the PriA-independent pathway of replication restart functions at forks that have been repaired to non-XOs.

Topological challenges to DNA replication: Conformations at the fork

The unwinding of the parental DNA duplex during replication causes a positive linking number difference, or superhelical strain, to build up around the elongating replication fork. The branching at the fork and this strain bring about different conformations from that of (−) supercoiled DNA that is not being replicated. The replicating DNA can form (+) precatenanes, in which the daughter DNAs are intertwined, and (+) supercoils. Topoisomerases have the essential role of relieving the superhelical strain by removing these structures. Stalled replication forks of molecules with a (+) superhelical strain have the additional option of regressing, forming a four-way junction at the replication fork. This four-way junction can be acted on by recombination enzymes to restart replication. Replication and chromosome folding are made easier by topological domain barriers, which sequester the substrates for topoisomerases into defined and concentrated regions. Domain barriers also allow replicated DNA to be (−) supercoiled. We discuss the importance of replicating DNA conformations and the roles of topoisomerases, focusing on recent work from our laboratory.

Rescue of stalled replication forks by RecG: Simultaneous translocation on the leading and lagging strand templates supports an active DNA unwinding model of fork reversal and Holliday junction formation

Modification of damaged replication forks is emerging as a crucial factor for efficient chromosomal duplication and the avoidance of genetic instability. The RecG helicase of Escherichia coli, which is involved in recombination and DNA repair, has been postulated to act on stalled replication forks to promote replication restart via the formation of a four-stranded (Holliday) junction. Here we show that RecG can actively unwind the leading and lagging strand arms of model replication fork structures in vitro. Unwinding is achieved in each case by simultaneous interaction with and translocation along both the leading and lagging strand templates at a fork. Disruption of either of these interactions dramatically inhibits unwinding of the opposing duplex arm. Thus, RecG translocates simultaneously along two DNA strands, one with 5′-3′ and the other with 3′-5′ polarity. The unwinding of both nascent strands at a damaged fork, and their subsequent annealing to form a Holliday junction, may explain the ability of RecG to promote replication restart. Moreover, the preferential binding of partial forks lacking a leading strand suggests that RecG may have the ability to target stalled replication intermediates in vivo in which lagging strand synthesis has continued beyond the leading strand.

Formation of Holliday junctions by regression of nascent DNA in intermediates containing stalled replication forks: RecG stimulates regression even when the DNA is negatively supercoiled

Replication forks formed at bacterial origins often encounter template roadblocks in the form of DNA adducts and frozen protein–DNA complexes, leading to replication-fork stalling and inactivation. Subsequent correction of the corrupting template lesion and origin-independent assembly of a new replisome therefore are required for survival of the bacterium. A number of models for replication-fork restart under these conditions posit that nascent strand regression at the stalled fork generates a Holliday junction that is a substrate for subsequent processing by recombination and repair enzymes. We show here that early replication intermediates containing replication forks stalled in vitro by the accumulation of excess positive supercoils could be cleaved by the Holliday junction resolvases RusA and RuvC. Cleavage by RusA was inhibited by the presence of RuvA and was stimulated by RecG, confirming the presence of Holliday junctions in the replication intermediate and supporting the previous proposal that RecG could catalyze nascent strand regression at stalled replication forks. Furthermore, RecG promoted Holliday junction formation when replication intermediates in which the replisome had been inactivated were negatively supercoiled, suggesting that under intracellular conditions, the action of RecG, or helicases with similar activities, is necessary for the catalysis of nascent strand regression.

The tight linkage between DNA replication and double-strand break repair in bacteriophage T4

Double-strand break (DSB) repair and DNA replication are tightly linked in the life cycle of bacteriophage T4. Indeed, the major mode of phage DNA replication depends on recombination proteins and can be stimulated by DSBs. DSB-stimulated DNA replication is dramatically demonstrated when T4 infects cells carrying two plasmids that share homology. A DSB on one plasmid triggered extensive replication of the second plasmid, providing a useful model for T4 recombination-dependent replication (RDR). This system also provides a view of DSB repair in T4-infected cells and revealed that the DSB repair products had been replicated in their entirety by the T4 replication machinery. We analyzed the detailed structure of these products, which do not fit the simple predictions of any of three models for DSB repair. We also present evidence that the T4 RDR system functions to restart stalled or inactivated replication forks. First, we review experiments involving antitumor drug-stabilized topoisomerase cleavage complexes. The results suggest that forks blocked at cleavage complexes are resolved by recombinational repair, likely involving RDR. Second, we show here that the presence of a T4 replication origin on one plasmid substantially stimulated recombination events between it and a homologous second plasmid that did not contain a T4 origin. Furthermore, replication of the second plasmid was increased when the first plasmid contained the T4 origin. Our interpretation is that origin-initiated forks become inactivated at some frequency during replication of the first plasmid and are then restarted via RDR on the second plasmid.

Roles of DNA polymerases V and II in SOS-induced error-prone and error-free repair in Escherichia coli

DNA polymerase V, composed of a heterotrimer of the DNA damage-inducible UmuC and UmuD\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} \begin{equation*}{\mathrm{_{2}^{^{\prime}}}}\end{equation*}\end{document} proteins, working in conjunction with RecA, single-stranded DNA (ssDNA)-binding protein (SSB), β sliding clamp, and γ clamp loading complex, are responsible for most SOS lesion-targeted mutations in Escherichia coli, by catalyzing translesion synthesis (TLS). DNA polymerase II, the product of the damage-inducible polB (dinA ) gene plays a pivotal role in replication-restart, a process that bypasses DNA damage in an error-free manner. Replication-restart takes place almost immediately after the DNA is damaged (≈2 min post-UV irradiation), whereas TLS occurs after pol V is induced ≈50 min later. We discuss recent data for pol V-catalyzed TLS and pol II-catalyzed replication-restart. Specific roles during TLS for pol V and each of its accessory factors have been recently determined. Although the precise molecular mechanism of pol II-dependent replication-restart remains to be elucidated, it has recently been shown to operate in conjunction with RecFOR and PriA proteins.

Intrinsic transcript cleavage activity of RNA polymerase.

The GreA and GreB transcript cleavage factors of Escherichia coli suppress elongation arrest and may have a proofreading role in transcription. With the use of E. coli greA-greB- mutant, RNA polymerase is demonstrated to possess substantial intrinsic transcript cleavage activity. Mildly alkaline pH mimics the effect of the Gre proteins by inducing transcript cleavage in ternary complexes and antagonizing elongation arrest through a cleavage-and-restart reaction. Thus, transcript cleavage constitutes the second enzymological activity of RNA polymerase along with polymerization/pyrophosphorolysis of RNA, whereas the Gre proteins merely enhance this intrinsic property.

Capacity-building through English language training to improve long-term resettlement success: A case study of the refugees along the Thailand-Burma border

Achieving long-term resettlement success is a challenge for many refugees seeking to restart their lives after displacement and being uprooted from their lives. Refugees must deal with finding employment, integrating into a society immensely different from what they have known their whole lives, and starting over from scratch. Learning a new language enables refugees to progress towards integration and long-term resettlement success, however, resettled refugees face a multitude of barriers in the U.S. to accessing language classes and attaining English proficiency. This study seeks to bridge this problem by exploring the possibilities of implementing a standardized language training program in the refugee camps to better prepare refugees for resettlement. A case study of the refugees along the Thai-Burma border demonstrated the significance of learning English in the camps on eventual English proficiency as well as the need for increased partnerships to overcome the barriers of lack of motivation and lack of funding. The author explores the possibilities of implementing a language training program in the camps by determining need, interest, barriers, and perceptions through the use of interviews, surveys, and focus groups of camp refugees, resettled refugees, and key organizational representatives. The significance of these results offers the possibility of leveraging and unlocking resettlement as a durable solution for more of the world's refugees in protracted situations.

An application of fuzzy logic for hardware/software partitioning in embedded systems

Hardware/Software partitioning (HSP) is a key task for embedded system co-design. The main goal of this task is to decide which components of an application are to be executed in a general purpose processor (software) and which ones, on a specific hardware, taking into account a set of restrictions expressed by metrics. In last years, several approaches have been proposed for solving the HSP problem, directed by metaheuristic algorithms. However, due to diversity of models and metrics used, the choice of the best suited algorithm is an open problem yet. This article presents the results of applying a fuzzy approach to the HSP problem. This approach is more flexible than many others due to the fact that it is possible to accept quite good solutions or to reject other ones which do not seem good. In this work we compare six metaheuristic algorithms: Random Search, Tabu Search, Simulated Annealing, Hill Climbing, Genetic Algorithm and Evolutionary Strategy. The presented model is aimed to simultaneously minimize the hardware area and the execution time. The obtained results show that Restart Hill Climbing is the best performing algorithm in most cases.

Partición hardware software de un codificador JPEG utilizando escalador de colinas estocástico

La partición hardware/software es una etapa clave dentro del proceso de co-diseño de los sistemas embebidos. En esta etapa se decide qué componentes serán implementados como co-procesadores de hardware y qué componentes serán implementados en un procesador de propósito general. La decisión es tomada a partir de la exploración del espacio de diseño, evaluando un conjunto de posibles soluciones para establecer cuál de estas es la que mejor balance logra entre todas las métricas de diseño. Para explorar el espacio de soluciones, la mayoría de las propuestas, utilizan algoritmos metaheurísticos; destacándose los Algoritmos Genéticos, Recocido Simulado. Esta decisión, en muchos casos, no es tomada a partir de análisis comparativos que involucren a varios algoritmos sobre un mismo problema. En este trabajo se presenta la aplicación de los algoritmos: Escalador de Colinas Estocástico y Escalador de Colinas Estocástico con Reinicio, para resolver el problema de la partición hardware/software. Para validar el empleo de estos algoritmos se presenta la aplicación de este algoritmo sobre un caso de estudio, en particular la partición hardware/software de un codificador JPEG. En todos los experimentos es posible apreciar que ambos algoritmos alcanzan soluciones comparables con las obtenidas por los algoritmos utilizados con más frecuencia.

Déficience dans la réparation par excision de nucléotides en phase S induite par la séquestration du facteur de réplication RPA

Introduction Les lésions induites par les rayons UV peuvent causer des blocages dans la réplication de l'ADN. Ces dommages sont éliminés par le processus moléculaire très conservé de réparation par excision de nucléotides (NER). Nous avons précédemment démontré que la protéine ATR, une kinase majeure impliquée dans le stress réplicatif, est requise pour une NER efficace, et ce exclusivement durant la phase S. Des résultats subséquents ont suggéré que ce prérequis n’était pas lié à la réponse induite par ATR, mais plutôt d’une conséquence globale causée par la présence de stress réplicatif. En ce sens, nous mettons l’emphase qu’après irradiation UV, le complexe RPA joue un rôle crucial dans l'activation des mécanismes de NER ainsi que dans le redémarrage des fourches de réplication bloquées. Hypothèses: En général, les mutations qui confèrent une augmentation du stress réplicatif engendrent une séquestration excessive du facteur RPA aux fourches de réplication bloquées ce qui réduit son accessibilité pour le NER. Méthodes et résultats: Le modèle de la levure a été choisi pour vérifier cette hypothèse. Nous avons développé un essai de NER spécifique à chacune des phases du cycle cellulaire pour démontrer que les cellules déficientes en Mec1, l’homologue d’ATR, sont défectives dans la réparation par excision de nucléotides spécifiquement en phase S. De plus, plusieurs autres mutants de levure, caractérisés par un niveau de dommages spontanés élevé, ont aussi exhibé un défaut similaire. Ces mutants ont démontré une fréquence et une intensité de formation de foyers de RPA plus élevée. Finalement, une diminution partielle de RPA dans les levures a induit un défaut significatif dans le NER spécifiquement durant la phase S. Conclusion: Nos résultats supportent la notion que la séquestration de RPA aux fourches de réplication endommagées durant la phase S prévient son utilisation pour la réparation par excision de nucléotides ce qui inhibe fortement l'efficacité de réparation. Cette étude chez la levure facilite l’élucidation du phénomène analogue chez l’humain et, ultimement, comprend des implications majeures dans la compréhension du mécanisme de développement des cancers UV-dépendants.

Transnistria – Where to? EPC Policy Brief, 13 March 2014

At the Vilnius summit of the Eastern Partnership in November 2013, Moldova initialled its Association Agreement with the EU, including a Deep and Comprehensive Free Trade Area (DCFTA) agreement, and is expected to sign the documents before the end of August 2014. Meanwhile, Russia has increased its presence and pressure in the region, as a consequence of which Armenia declined the AA and DCFTA with the EU and Ukraine, after months of protests and political paralysis, now has part of its territory occupied by Russia. Moldova is no exception to Russian pressure. As the country gets closer to upgrading relations with the EU, Russia has increased its activities in Moldova, including in the autonomous region of Gagauzia and in the breakaway region of Transnistria. Even though the “5+2” negotiations for the settlement of the Transnistria conflict continue, the number of incidents in and around this region have increased. The window of opportunity created by the involvement of Germany in the settlement of the conflict and the restart of the “5+2” negotiations in late 2011 seems to have closed. Given the recent events in the region and in Moldova/Transnistria, including the potential impact of DCFTA and visa liberalisation, Chisinau finds it increasingly difficult to manage the juggling act between its EU commitment and dialogue with Tiraspol. This Policy Brief presents the background, state of play and prospects of the Transnistria conflict while also focusing on the potential impact of Moldova’s Association Agreement with the EU on the settlement of the conflict.