Mutagenesis and modelling of the alpha(1b)-adrenergic receptor highlight the role of the helix 3/helix 6 interface in receptor activation.

Computer simulations on a new model of the alpha1b-adrenergic receptor based on the crystal structure of rhodopsin have been combined with experimental mutagenesis to investigate the role of residues in the cytosolic half of helix 6 in receptor activation. Our results support the hypothesis that a salt bridge between the highly conserved arginine (R143(3.50)) of the E/DRY motif of helix 3 and a conserved glutamate (E289(6.30)) on helix 6 constrains the alpha1b-AR in the inactive state. In fact, mutations of E289(6.30) that weakened the R143(3.50)-E289(6.30) interaction constitutively activated the receptor. The functional effect of mutating other amino acids on helix 6 (F286(6.27), A292(6.33), L296(6.37), V299(6.40,) V300(6.41), and F303(6.44)) correlates with the extent of their interaction with helix 3 and in particular with R143(3.50) of the E/DRY sequence.

The Disadvantage of winning an election

This paper analyzes the problem that an incumbent faces during the legislature when deciding how to react to popular initiatives or policy proposals coming from different sources. We argue that this potential source of electoral disadvantage that the incumbent obtains after being elected can jeopardize the reelection possibilities of the incumbent. We analyze the decision of the incumbent when facing reelection and we characterize the conditions under which the advantages that the incumbent obtains can overcome the disadvantages. Finally, we use the results of this analysis to discuss some implications of the use of mechanisms of direct democracy like referenda and popular assemblies on electoral competition.

Multiple imputations applied to the DREAM3 phosphoproteomics challenge: a winning strategy.

DREAM is an initiative that allows researchers to assess how well their methods or approaches can describe and predict networks of interacting molecules [1]. Each year, recently acquired datasets are released to predictors ahead of publication. Researchers typically have about three months to predict the masked data or network of interactions, using any predictive method. Predictions are assessed prior to an annual conference where the best predictions are unveiled and discussed. Here we present the strategy we used to make a winning prediction for the DREAM3 phosphoproteomics challenge. We used Amelia II, a multiple imputation software method developed by Gary King, James Honaker and Matthew Blackwell[2] in the context of social sciences to predict the 476 out of 4624 measurements that had been masked for the challenge. To chose the best possible multiple imputation parameters to apply for the challenge, we evaluated how transforming the data and varying the imputation parameters affected the ability to predict additionally masked data. We discuss the accuracy of our findings and show that multiple imputations applied to this dataset is a powerful method to accurately estimate the missing data. We postulate that multiple imputations methods might become an integral part of experimental design as a mean to achieve cost savings in experimental design or to increase the quantity of samples that could be handled for a given cost.

Combined simulation and mutagenesis analyses reveal the involvement of key residues for peroxisome proliferator-activated receptor alpha helix 12 dynamic behavior.

The dynamic properties of helix 12 in the ligand binding domain of nuclear receptors are a major determinant of AF-2 domain activity. We investigated the molecular and structural basis of helix 12 mobility, as well as the involvement of individual residues with regard to peroxisome proliferator-activated receptor alpha (PPARalpha) constitutive and ligand-dependent transcriptional activity. Functional assays of the activity of PPARalpha helix 12 mutants were combined with free energy molecular dynamics simulations. The agreement between the results from these approaches allows us to make robust claims concerning the mechanisms that govern helix 12 functions. Our data support a model in which PPARalpha helix 12 transiently adopts a relatively stable active conformation even in the absence of a ligand. This conformation provides the interface for the recruitment of a coactivator and results in constitutive activity. The receptor agonists stabilize this conformation and increase PPARalpha transcription activation potential. Finally, we disclose important functions of residues in PPARalpha AF-2, which determine the positioning of helix 12 in the active conformation in the absence of a ligand. Substitution of these residues suppresses PPARalpha constitutive activity, without changing PPARalpha ligand-dependent activation potential.

An amphipathic alpha-helix at the C terminus of hepatitis C virus nonstructural protein 4B mediates membrane association.

Nonstructural protein 4B (NS4B) plays an essential role in the formation of the hepatitis C virus (HCV) replication complex. It is an integral membrane protein that has been only poorly characterized to date. It is believed to comprise a cytosolic N-terminal part, a central part harboring four transmembrane passages, and a cytosolic C-terminal part. Here, we describe an amphipathic alpha-helix at the C terminus of NS4B (amino acid residues 229 to 253) that mediates membrane association and is involved in the formation of a functional HCV replication complex.

People on the Edge See More. A conversation with the cuban-american writer Cristina García

L'entrevista amb l'escriptora cubano-americana Cristina García explora el tema de la identitat cubanoamericana i desvetlla la riquesa literària que sorgeix de la fusió de dues cultures, la cubana i la nord-americana, i com aquesta fusió innova la literatura nord-americana tradicional. En la seva novel·la, Dreaming in Cuban (1992), l'escriptora explora els efectes de la Revolució castrista des de la perspectiva de les dones cubanes que van quedar-se a l'illa, així com de les dones que emigraren als Estats Units. The conversation with Cuban-American writer Cristina García explores what it means to be Cuban-American, and reveals how to grow bicultural enriches mainstream American literature. In her novel Dreaming in Cuban (1992), the writer explores the effects of the Castro Revolution from the perspective of Cuban women who remained in Cuba, as well as from the experience of women who emigrated to the United States.

People on the Edge See More. A conversation with the cuban-american writer Cristina García

L'entrevista amb l'escriptora cubano-americana Cristina García explora el tema de la identitat cubanoamericana i desvetlla la riquesa literària que sorgeix de la fusió de dues cultures, la cubana i la nord-americana, i com aquesta fusió innova la literatura nord-americana tradicional. En la seva novel·la, Dreaming in Cuban (1992), l'escriptora explora els efectes de la Revolució castrista des de la perspectiva de les dones cubanes que van quedar-se a l'illa, així com de les dones que emigraren als Estats Units. The conversation with Cuban-American writer Cristina García explores what it means to be Cuban-American, and reveals how to grow bicultural enriches mainstream American literature. In her novel Dreaming in Cuban (1992), the writer explores the effects of the Castro Revolution from the perspective of Cuban women who remained in Cuba, as well as from the experience of women who emigrated to the United States.

Methionine-321 in the C-terminal alpha-helix of catabolic ornithine carbamoyltransferase from Pseudomonas aeruginosa is important for positive homotropic cooperativity.

Pseudomonas aeruginosa has a pair of distinct ornithine carbamoyltransferases. The anabolic ornithine carbamoyltransferase encoded by the argF gene catalyzes the formation of citrulline from ornithine and carbamoylphosphate. The catabolic ornithine carbamoyltransferase encoded by the arcB gene promotes the reverse reaction in vivo; although this enzyme can be assayed in vitro for citrulline synthesis, its unidirectionality in vivo is determined by its high concentration at half maximum velocity for carbamoylphosphate ([S]0.5) and high cooperativity toward this substrate. We have isolated mutant forms of catabolic ornithine carbamoyltransferase catalyzing the anabolic reaction in vivo. The corresponding arcB mutant alleles on a multicopy plasmid specifically suppressed an argF mutation of P. aeruginosa. Two new mutant enzymes were obtained. When methionine 321 was replaced by isoleucine, the mutant enzyme showed loss of homotropic cooperativity at physiological carbamoylphosphate concentrations. Substitution of glutamate 105 by lysine resulted in a partial loss of the sigmoidal response to increasing carbamoylphosphate concentrations. However, both mutant enzymes were still sensitive to the allosteric activator AMP and to the inhibitor spermidine. These results indicate that at least two residues of catabolic ornithine carbamoyltransferase are critically involved in positive carbamoylphosphate cooperativity: glutamate 105 (previously known to be important) and methionine 321. Mutational changes in either amino acid will affect the geometry of helix H2, which contains several residues required for carbamoylphosphate binding.

Amphipathic alpha-helix AH2 is a major determinant for the oligomerization of hepatitis C virus nonstructural protein 4B.

Nonstructural protein 4B (NS4B) is a key organizer of hepatitis C virus (HCV) replication complex formation. It induces a specific membrane rearrangement, designated membranous web, that serves as a scaffold for the HCV replication complex. However, the mechanisms underlying membranous web formation are poorly understood. Based on fluorescence resonance energy transfer (FRET) and confirmatory coimmunoprecipitation analyses, we provide evidence for an oligomerization of NS4B in the membrane environment of intact cells. Several conserved determinants were found to be involved in NS4B oligomerization, through homotypic and heterotypic interactions. N-terminal amphipathic ?-helix AH2, comprising amino acids 42 to 66, was identified as a major determinant for NS4B oligomerization. Mutations that affected the oligomerization of NS4B disrupted membranous web formation and HCV RNA replication, implying that oligomerization of NS4B is required for the creation of a functional replication complex. These findings enhance our understanding of the functional architecture of the HCV replication complex and may provide new angles for therapeutic intervention. At the same time, they expand the list of positive-strand RNA virus replicase components acting as oligomers.

Tindara Addabbo, Marie-Pierre Arrizabalaga, Cristina Borderías y Alastair Owens (eds.), Gender Inequalities, Households and the Production of Well-being in Modern Europe

Ressenya del llibre: 'Gender Inequalities, Households and the Production of Well-being in Modern Europe'

Winning time in land warfare : a reading of the classical strategists prior to the age of mechanization

We live in an age where rationalization and demands of efficiency taint every aspect of our lives both as individuals and as a society. Even warfare cannot escape the increased speed of human interaction. Time is a resource to be managed. It has to be optimized, saved and won in military affairs as well. The purpose of this research paper is to analyze the dogmatic texts of military thought to search for answers what the classics of strategy saw in the interrelations of temporality and warfare and if their thoughts remain meaningful in the contemporary conjunction. Since the way a society functions is reflected in the way it conducts its wars, there naturally are differences between an agrarian, industrial and information society. Theorists of different eras emphasize things specific to their times, but warfare, like any human interaction, is always bounded by temporality. Not only is the pace of warfare dependent on the progress of the society, but time permeates warfare in all its aspects. This research paper focuses on two specific topics that arose from the texts themselves; how should time be managed and manipulated in warfare and how to economize and “win” it from the enemy. A method where lengthy quotations are used to illustrate the main point of the strategists has been chosen for this research paper. While Clausewitz is the most prominent source of quotations, thoughts from ancient India and China are represented as well to prove that the combination of right force in the right place at the right time is still the way of the victorious. Tactics change in the course of time but the principles of strategy remain unaltered and are only adapted to suit new situations. While ancient and pre-modern societies had their focus on finding auspicious moments for battle in the flow of kronos-time based on divinities, portents and auguries, we can trace elements of manipulation of time in warfare from the earliest surviving texts. While time as a fourth dimension of the battlespace emerged only in the modern era, all through the history of military thought it has had a profound meaning. In the past time could be squandered, today it always has to be won. This paper asks the question “why”.

Redefining U.S. borders : a reading of Sandra Cisneros’s Caramelo, Cristina Garcia’s The Agüero Sisters, and David Plante’s The Family and The Native

Cette thèse propose une redéfinition de la notion de frontière dans le contexte américain, avec pour point de départ les romans de trois voix littéraires issues de trois minorités ethniques : Sandra Cisneros (Caramelo), Cristina Garcia (The Agüero Sisters) et David Plante (The Family et The Native). Je conceptualise la frontière comme fluctuation entre mouvement et immobilité, entre porosité et imperméabilité. Dans le premier chapitre, je fournis des repères sur la théorie des frontières et j'analyse les avancées de ce champ d'étude, du concept de terre frontalière (“Borderland Theories”) jusqu'aux récits d'immigration. Je propose un cadre conceptuel que j'appelle « Écrire la frontière à partir de la perspective de la frontière », lequel permet une lecture neuve des récits de frontière, et une redéfinition de la notion elle-même. Prise comme perspective, la frontière est une dynamique vivante, ce qui la rend plurielle et impossible à fixer définitivement; aussi les récits de frontière présentent-ils une grande variété d’expériences, toutes liées à des moments et à des points de vue uniques. Dans le second chapitre, j’analyse la porosité des frontières dans le contexte géopolitique contemporain, en mettant en lumière comment la colonisation, la mondialisation économique et l’immigration sont autant de mécanismes de transgression des frontières qui suivent des orientations transnationales, dénationales et postnationales. Dans le troisième chapitre, j’étudie la résurgence des frontières dans la vie des immigrants qui habitent aux États-Unis. J’identifie l’insécurité capitaliste ainsi que la marchandisation de l’espace et de l’ethnicité comme étant à l'origine du renforcement des frontières délimitant les quartiers ethniques; génératrices de stéréotypes négatifs, ces divisions physiques deviennent une technologie d’exclusion et d’injustice sociale. Le dernier chapitre présente une lecture des aspects esthétiques de la frontière, voyant comment ils peuvent contribuer à écrire la frontière à partir de la perspective de la frontière. Dans les textes à l'étude, j'examine de près la problématisation du concept de représentation, la multiplicité des points de vue narratifs, l’inaccessibilité du réel, et la partialité de la médiation. Mots clés : Théories et écrits sur les frontières, minorités ethniques aux États-Unis, multiculturalisme, culture, immigration, mondialisation, espace, place, territoire, état-nation, nationalisme, histoire, langue et langage, représentation, communauté, justice sociale, citoyenneté