Lagging strand replication of rolling-circle plasmids: Specific recognition of the ssoA-type origins in different gram-positive bacteria

Many bacterial plasmids replicate by a rolling-circle mechanism that involves the generation of single-stranded DNA (ssDNA) intermediates. Replication of the lagging strand of such plasmids initiates from their single strand origin (sso). Many different types of ssos have been identified. One group of ssos, termed ssoA, which have conserved sequence and structural features, function efficiently only in their natural hosts in vivo. To study the host specificity of sso sequences, we have analyzed the functions of two closely related ssoAs belonging to the staphylococcal plasmid pE194 and the streptococcal plasmid pLS1 in Staphylococcus aureus. The pLS1 ssoA functioned poorly in vivo in S. aureus as evidenced by accumulation of high levels of ssDNA but supported efficient replication in vitro in staphylococcal extracts. These results suggest that one or more host factors that are present in sufficient quantities in S. aureus cell-free extracts may be limiting in vivo. Mapping of the initiation points of lagging strand synthesis in vivo and in vitro showed that DNA synthesis initiates from specific sites within the pLS1 ssoA. These results demonstrate that specific initiation of replication can occur from the pLS1 ssoA in S. aureus although it plays a minimal role in lagging strand synthesis in vivo. Therefore, the poor functionality of the pLS1 in vivo in a nonnative host is caused by the low efficiency rather than a lack of specificity of the initiation process. We also have identified ssDNA promoters and mapped the primer RNAs synthesized by the S. aureus and Bacillus subtilis RNA polymerases from the pE194 and pLS1 ssoAs. The S. aureus RNA polymerase bound more efficiently to the native pE194 ssoA as compared with the pLS1 ssoA, suggesting that the strength of RNA polymerase–ssoA interaction may play a major role in the functionality of the ssoA sequences in Gram-positive bacteria.

Nonuniformity of the constitutive law parameters for shear rupture and quasistatic nucleation to dynamic rupture: a physical model of earthquake generation processes.

Based on the recent high-resolution laboratory experiments on propagating shear rupture, the constitutive law that governs shear rupture processes is discussed in view of the physical principles and constraints, and a specific constitutive law is proposed for shear rupture. It is demonstrated that nonuniform distributions of the constitutive law parameters on the fault are necessary for creating the nucleation process, which consists of two phases: (i) a stable, quasistatic phase, and (ii) the subsequent accelerating phase. Physical models of the breakdown zone and the nucleation zone are presented for shear rupture in the brittle regime. The constitutive law for shear rupture explicitly includes a scaling parameter Dc that enables one to give a common interpretation to both small scale rupture in the laboratory and large scale rupture as earthquake source in the Earth. Both the breakdown zone size Xc and the nucleation zone size L are prescribed and scaled by Dc, which in turn is prescribed by a characteristic length lambda c representing geometrical irregularities of the fault. The models presented here make it possible to understand the earthquake generation process from nucleation to unstable, dynamic rupture propagation in terms of physics. Since the nucleation process itself is an immediate earthquake precursor, deep understanding of the nucleation process in terms of physics is crucial for the short-term (or immediate) earthquake prediction.

Xce haplotypes show modified methylation in a region of the active X chromosome lying 3' to Xist.

During early mammalian embryogenesis, one of the two X chromosomes in somatic cells of the female becomes inactivated through a process that is thought to depend on a unique initiator region, the X-chromosome inactivation center (Xic). The recently characterized Xist sequence (X-inactive-specific transcript) is thought to be a possible candidate for Xic. In mice a further genetic element, the X chromosome-controlling element (Xce), is also known to influence the choice of which of the two X chromosomes is inactivated. We report that a region of the mouse X chromosome lying 15 kb distal to Xist contains several sites that show hypermethylation specifically associated with the active X chromosome. Analysis of this region in various Xce strains has revealed a correlation between the strength of the Xce allele carried and the methylation status of this region. We propose that such a region could be involved in the initial stages of the inactivation process and in particular in the choice of which of the two X chromosomes present in a female cell will be inactivated.

Once Upon a Time in Therapy: Crafting Therapeutic Narratives to Help Familiarize Children with the Therapeutic Process.

Young children often harbor misconceptions about psychotherapy and the role of psychologists. These misconceptions are ignited by rumors and misinformation that are provided to the child by a variety of sources and can compromise both the effectiveness of therapy and the therapeutic dyad. In this paper we explore how recent trends in patient engagement in child psychotherapy, cultural dynamics between patients and practitioners, and children's lack of knowledge surrounding mental health services can negatively impact therapy. Wednesday Afternoons with Dr. J. (WADJ) is a whimsical fictional therapeutic narrative created to inform children about aspects of the therapeutic process while providing adults with tangible structure surrounding how to talk to children about mental healthcare. The advantages of utilizing this narrative to prime children for therapy are discussed, as are methods for promoting the narrative to the greater community.

Arthroscopic stabilisation of an acute acromioclavicular dislocation grade III in a patient with ectopic insertion of the pectoralis minor: technical considerations

The different approaches used in arthroscopic stabilisation of the acromioclavicular joint are well known. However, and despite a great incidence of ectopic pectoralis minor insertion, an alternative choice for the use of arthroscopic portal has not being sufficiently described. Here, we describe a case of acute acromioclavicular dislocation grade III. The arthroscopic stabilisation was achieved using the TightRope (Arthrex, Naples, USA) implant. Through this technique, the approach to the articular portion of the coracoid process can be made intra-articularly or from the subacromial space. We accessed intra-articularly, by opening the rotator interval to reach the coracoid process from the joint cavity. After opening the rotator interval, an ectopic insertion of the pectoralis minor was observed. The choice of approach of the coracoid process from the subacromial space would have complicated the intervention, making it necessary to sever the ectopic tendon to complete the technique, lengthening the surgical time and increasing the chance of complications. For this reason, the use of a standard posterior portal providing intra-articular arthroscopic access through the rotator interval is recommended since the aforementioned anatomical variation is not infrequent. Level of evidence Therapeutic studies—investigating the results of treatment, Level V.

Comment Letter: The NYSE and the End of the Structural Separation between Regulatory and Commercial Interests of the Exchange

The NYSE transformed into a for profit entity in 2006. As part of the approval process, the NYSE agreed to structurally separate the regulatory function from the business function. In doing so, the NYSE created NYSE Regulation, a non-profit with an independent board, to handle most regulatory matters. During the comment period, a spirited debate arose over the ability of a for profit company to carry out a regulatory mission. Some suggested that the regulatory function was incompatible with a "for profit" motive and that NYSE Regulation should be spun off. Others accepted the proposed structure but called for additional changes designed to reduce the possible influence of the public holding company over the regulatory function. In the end, the SEC approved the structure but with a number of prophylactic safeguards including the requirement that NYSE Regulation have a board consisting of all independent directors (save the CEO) and that directors from the for profit holding company could not make up a majority of the board. More recently, however, the NYSE has proposed to end the structural separation of the two functions and instead put in place a functional separation. The proposal would result in the termination of the delegation agreement between the Exchange and NYSE Regulation and the creation of both a Regulatory Oversight Committee of the Board of Directors of the Exchange and the creation of a Chief Regulatory Officer. This letter examines the history of the separation of the two functions and critiques the NYSE's proposal.

Nostalgic representation of reality in television fiction: an empirical study based on the analysis of the Spanish television series Cuéntame cómo pasó

The Spanish transition from dictatorship to democracy is often described as an example of negotiation or agreement between the elites (Sánchez Cuenca and Aguilar, 2009: 433). Journalistic and political elites, aware of their important historical role, agreed a consensus on certain issues (democracy, constitution, amnesty) or characters (King Juan Carlos I), in order to ensure the stability of the democratic process (Zugasti, 2007, 2008). Television, which articulates the discourse of the masses, has been one of the basic means used to illustrate the development. Among the highlights of recent major audiovisual content, Cuéntame cómo pasó (2001-present) -a TV-series designed to explain changes with a nostalgic tone in Spanish society since 1968 until today- stands out. By choosing a random sample of episodes for this research we propose to verify the validity of the representation of the political process which contextualizes the series. By analyzing many elements, such as the opinions of the main characters, their personal, political and geographical situations, we try to show the construction of a focal point that sanctifies the official version. We also stress the pacifying and nostalgic tone, which constructs stereotypes and taboos about the process and which characterizes this series as a symbolic culmination of the democratization undertaken by the elites.

Calibration of the length of a chain of single gold atoms

Using a scanning tunnelling microscope or mechanically controllable break junction it has been shown that it is possible to control the formation of a wire made of single gold atoms. In these experiments an interatomic distance between atoms in the chain of ∼3.6 Å was reported which is not consistent with recent theoretical calculations. Here, using precise calibration procedures for both techniques, we measure the length of the atomic chains. Based on the distance between the peaks observed in the chain length histogram we find the mean value of the interatomic distance before chain rupture to be 2.5±0.2 Å. This value agrees with the theoretical calculations for the bond length. The discrepancy with the previous experimental measurements was due to the presence of He gas, that was used to promote the thermal contact, and which affects the value of the work function that is commonly used to calibrate distances in scanning tunnelling microscopy and mechanically controllable break junctions at low temperatures.

Effect of the porous structure in carbon materials for CO2 capture at atmospheric and high-pressure

Activated carbons prepared from petroleum pitch and using KOH as activating agent exhibit an excellent behavior in CO2 capture both at atmospheric (∼168 mg CO2/g at 298 K) and high pressure (∼1500 mg CO2/g at 298 K and 4.5 MPa). However, an exhaustive evaluation of the adsorption process shows that the optimum carbon structure, in terms of adsorption capacity, depends on the final application. Whereas narrow micropores (pores below 0.6 nm) govern the sorption behavior at 0.1 MPa, large micropores/small mesopores (pores below 2.0–3.0 nm) govern the sorption behavior at high pressure (4.5 MPa). Consequently, an optimum sorbent exhibiting a high working capacity for high pressure applications, e.g., pressure-swing adsorption units, will require a poorly-developed narrow microporous structure together with a highly-developed wide microporous and small mesoporous network. The appropriate design of the preparation conditions gives rise to carbon materials with an extremely high delivery capacity ∼1388 mg CO2/g between 4.5 MPa and 0.1 MPa. Consequently, this study provides guidelines for the design of carbon materials with an improved ability to remove carbon dioxide from the environment at atmospheric and high pressure.

Thermal decomposition of the different particles size fractions of almond shells and olive stones. Thermal behaviour changes due to the milling processes

The aim of this study is to investigate the effect of particle size on the non-isothermal pyrolysis of almond shells (AS) and olive stones (OS) and to show possible differences in the composition of the different fractions obtained after milling and sieving. The results obtained from the study of different particle size of AS and OS samples show significant differences in the solid residue obtained and in the shape and overlapping degree of the peaks, especially with the smaller particle size. These differences can be due to different factors: (a) the amount of inorganic matter, which increases as particle size decreases, (b) heat and mass transfer processes, (c) different sample composition as a consequence of the milling process which may provoke changes in the structure and the segregation of the components (in addition to the ashes) increasingly changes the composition of the sample as the particle size decreases.

Tourism potential analysis of the castles of Vinalopó, Spain: an opportunity for social and tourist development of cultural heritage

Within the overall framework of the renewal process of coastal tourist destinations, cultural heritage has frequently been used as a key argument for the introduction and development of strategies for the diversification and differentiation of the traditional tourist product based on sun and sand. This is the situation of the province of Alicante, identified with the Costa Blanca geotourism brand, where there are important economic issues that could contribute to the renewal of this coastal tourist destination. One of the most significant heritage values of this space consists of a series of medieval fortresses located along the axis of the Vinalopó River, which has acted since prehistoric times as a natural route from within the provincial mainland to the coast. It is precisely the argument of this historical, territorial and landscape content that has been used repeatedly in recent years to develop initiatives aimed at the creation of a tourist product, currently inexistent, based on the route of the castles of Vinalopó. This communication aims to analyse the degree of tourism potential of the fortresses located in the towns of Biar, Banyeres de Mariola, Sax, Villena, Novelda, Elda, Petrer and Elche, which constitute the core of municipalities where these medieval fortresses are located, finally pointing out some proposals for the creation of a heritage tourism product.

Phase progression throughout the therapeutic process : longitudinal case studies

Tese de doutoramento, Psicologia (Psicologia Clínica), Universidade de Lisboa, Faculdade de Psicologia, 2016

Do Institutional Changes Make a Difference? A veto player analysis of how institutional changes in the Council of the EU influence legislative efficiency and outputs. Brugges Political Research Paper No. 31, October 2013

A driving argument behind recent EU treaty reforms was that more qualified majority voting (QMV) was required to reduce the potential dangers of legislative paralysis caused by enlargement. Whilst existing literature on enlargement mostly focuses on the question of what changed in the legislative process after the 2004 enlargement, the question of why these changes occurred has been given far less attention. Through the use of a single veto player theoretical model, this paper seeks to test and explain whether enlargement reduces the efficiency of the legislative process and alters the type of legislation produced, and whether QMV can compensate for these effects. In doing this, it offers a theoretical explanation as to why institutional changes that alter the level of cohesion between actors in the Council have an influence over both the legislative process and its outcomes.

Eurasian integration. Russia's attempt at the economic unification of the post-Soviet area. OSW Study 44/2013

In 2009, Vladimir Putin, the then Russian prime minister, gave impetus to the establishment of closer relations within what was then a still narrow group of three countries: Russia, Kazakhstan and Belarus. Russia was determined in embarking on the implementation of the principles of the Customs Union among these three states and, since 2012, within the Common Economic Space as well. This process of integration is intended to bring about the introduction of ‘four freedoms’ in this area: the free movement of goods, services, capital and labour. From Moscow’s point of view, building up such integration structures is especially necessary in order to counteract the economic expansion of the European Union and China. It also feels it is important to take measures against the loosening of the bonds between the CIS countries and Russia. At the same time, close co-operation is expected to guarantee for Russia that the strong politico-economic influences in this area will be maintained. Despite the numerous limitations of the integration process, such as the small number of the participating states or limited progress in implementing the CES, this is still the most advanced integration programme in the region seen since the collapse of the USSR. Progress in putting the rules of the Customs Union into practice can be seen as a success for Moscow. In turn, the formation of the CES is still at an early stage, and it is difficult to determine at this point to what extent the three countries will harmonise their markets.

Gibt es eine politische Philosophie der Europäischen Union? = Is there a political philosophy of the European Union?. ZEI Discussion Paper C223, 2014

In a theoretical context, the European Union is generally interpreted through the prism of integration theories, which in turn reflect the ever changing empirical reality of the integration process. ZEI Director Ludger Kühnhardt asks if and to what extent the process of European integration has begun to generate a specific political philosophy which uses the EU - and not the classical notion of the state – as the starting and reference point for its reasoning. Kühnhardt examines examples – such as the European notion of civil rights and the notion of the Union itself, but also critical categories such as euroskepticism – which indicate that the EU itself is beginning to be the starting point and frame of reference for a reflection on the common good. For now, a political philosophy in the context of the European Union exists only in an embryonic stage, but the topic may generate intellectual insights through further and deeper research.