Effects of Dicer and Argonaute down-regulation on mRNA levels in human HEK293 cells

RNA interference and the microRNA (miRNA) pathway can induce sequence-specific mRNA degradation and/or translational repression. The human genome encodes hundreds of miRNAs that can post-transcriptionally repress thousands of genes. Using reporter constructs, we observed that degradation of mRNAs bearing sites imperfectly complementary to the endogenous let-7 miRNA is considerably stronger in human HEK293 than HeLa cells. The degradation did not result from the Ago2-mediated endonucleolytic cleavage but it was Dicer- and Ago2-dependent. We used this feature of HEK293 to address the size of a pool of transcripts regulated by RNA silencing in a single cell type. We generated HEK293 cell lines depleted of Dicer or individual Ago proteins. The cell lines were used for microarray analyses to obtain a comprehensive picture of RNA silencing. The 3'-untranslated region sequences of a few hundred transcripts that were commonly up-regulated upon Ago2 and Dicer knock-downs showed a significant enrichment of putative miRNA-binding sites. The up-regulation upon Ago2 and Dicer knock-downs was moderate and we found no evidence, at the mRNA level, for activation of silenced genes. Taken together, our data suggest that, independent of the effect on translation, miRNAs affect levels of a few hundred mRNAs in HEK293 cells

The changing patterns of group politics in Britain

Two interpretations of ways in which group politics in Britain have presented challenges to democracy are reviewed: neo-corporatism or pluralistic stagnation and the rise of single issue interest groups. The disappearance of the first paradigm created a political space for the second to emerge. A three-phase model of group activity is developed: a phase centred around production interests, followed by the development of broadly based 'other regarding' groups, succeeded by fragmented, inner directed groups focusing on particular interests. Explanations of the decay of corporatism are reviewed. Single issue group activity has increased as party membership has declined and is facilitated by changes in traditional media and the development of the internet. Such groups can overload the policy-making process and frustrate depoliticisation. Debates about the constitution and governance have largely ignored these issues and there is need for a debate

Post-Retained Single Crowns versus Fixed Dental Prostheses: A 7-Year Prospective Clinical Study

Biomechanical integrity of endodontically treated teeth (ETT) is often compromised. Degree of hard tissue loss and type of final prosthetic restoration should be carefully considered when making a treatment plan. The objective of this prospective clinical trial was to assess the influence of the type of prosthetic restoration as well as the degree of hard tissue loss on 7-y clinical performance of ETT restored with fiber posts. Two groups (n = 60) were defined depending on the type of prosthetic restoration needed: 1) single unit porcelain-fused-to-metal (PFM) crowns (SCs) and 2) 3- to 4-unit PFM fixed dental prostheses (FDPs), with 1 healthy and 1 endodontically treated and fiber post-restored abutment. Within each group, samples were divided into 2 subgroups (n = 30) according to the amount of residual coronal tissues after abutment buildup and final preparation: A) >50% of coronal residual structure or B) equal to or <50% of coronal residual structure. The clinical outcome was assessed based on clinical and intraoral radiographic examinations at the recalls after 6, 12, 24, 36, 48, and 84 mo. Data were analyzed by Kaplan-Meier log-rank test and Cox regression analysis (P < 0.05). The overall 7-y survival rate of ETT restored with fiber post and either SCs or FDPs was 69.2%. The highest 84-mo survival rate was recorded in group 1A (90%), whereas teeth in group 2B exhibited the lowest performance (56.7% survival rate). The log-rank test detected statistically significant differences in survival rates among the groups (P = 0.048). Cox regression analysis revealed that the amount of residual coronal structure (P = 0.041; hazard ratio [HR], 2.026; 95% confidence interval [CI] for HR, 1.031–3.982) and the interaction between the type of prosthetic restoration and the amount of residual coronal structure (P = 0.024; HR, 1.372; 95% CI for HR, 1.042–1.806) were statistically significant factors for survival (ClinicalTrials.gov NCT01532947)

Are language production problems apparent in adults who no longer meet diagnostic criteria for attention-deficit/hyperactivity disorder?

In this study, we examined sentence production in a sample of adults (N = 21) who had had attention-deficit/hyperactivity disorder (ADHD) as children, but as adults no longer met DSM-IV diagnostic criteria (APA, 2000). This “remitted” group was assessed on a sentence production task. On each trial, participants saw two objects and a verb. Their task was to construct a sentence using the objects as arguments of the verb. Results showed more ungrammatical and disfluent utterances with one particular type of verb (i.e., participle). In a second set of analyses, we compared the remitted group to both control participants and a “persistent” group, who had ADHD as children and as adults. Results showed that remitters were more likely to produce ungrammatical utterances and to make repair disfluencies compared to controls, and they patterned more similarly to ADHD participants. Conclusions focus on language output in remitted ADHD, and the role of executive functions in language production

Together forever? Explaining exclusivity in party-firm relations

Parties and firms are the key actors of representative democracy and capitalism respectively and the dynamic of attachment between them is a central feature of any political economy. This is the first article to systematically analyse the exclusivity of party-firm relations. We consider exclusivity at a point in time and exclusivity over time. Does a firm have a relationship with only one party at a given point in time, or is it close to more than one party? Does a firm maintain a relationship with only one party over time, or does it switch between parties? Most important, how do patterns of exclusivity impact on a firm’s ability to lobby successfully? We propose a general theory, which explains patterns of party-firm relations by reference to the division of institutions and the type of party competition in a political system. A preliminary test of our theory with Polish survey data confirms our predictions, establishing a promising hypothesis for future research

Trade unions and precariat in Europe : representative claims

Trade unions have been charged with neglecting labour market ‘outsiders’, while alternative actors have emerged to represent these. In response, unions have stepped up their claim to be representative of all workers, without distinction. We review the theoretical and policy debates on this issue, and argue that representation as such has been under-theorized. We draw on Saward’s concept of ‘representative claims’ to analyse the different grounds for competing assertions of representativeness. We identify four main forms of claims, and illustrate these with empirical examples. We conclude that these different claims are mutually reinforcing in stimulating attention to the outsiders, and in their interaction with institutional settings, they have a performative effect in defining new social actors

The Adaptor Molecule Nck Localizes the WAVE Complex to Promote Actin Polymerization during CEACAM3-Mediated Phagocytosis of Bacteria

Background: CEACAM3 is a granulocyte receptor mediating the opsonin-independent recognition and phagocytosis of human-restricted CEACAM-binding bacteria. CEACAM3 function depends on an intracellular immunoreceptor tyrosine-based activation motif (ITAM)-like sequence that is tyrosine phosphorylated by Src family kinases upon receptor engagement. The phosphorylated ITAM-like sequence triggers GTP-loading of Rac by directly associating with the guanine nucleotide exchange factor (GEF) Vav. Rac stimulation in turn is critical for actin cytoskeleton rearrangements that generate lamellipodial protrusions and lead to bacterial uptake. Principal Findings: In our present study we provide biochemical and microscopic evidence that the adaptor proteins Nck1 and Nck2, but not CrkL, Grb2 or SLP-76, bind to tyrosine phosphorylated CEACAM3. The association is phosphorylation-dependent and requires the Nck SH2 domain. Overexpression of the isolated Nck1 SH2 domain, RNAi-mediated knock-down of Nck1, or genetic deletion of Nck1 and Nck2 interfere with CEACAM3-mediated bacterial internalization and with the formation of lamellipodial protrusions. Nck is constitutively associated with WAVE2 and directs the actin nucleation promoting WAVE complex to tyrosine phosphorylated CEACAM3. In turn, dominant-negative WAVE2 as well as shRNA-mediated knock-down of WAVE2 or the WAVE-complex component Nap1 reduce internalization of bacteria. Conclusions: Our results provide novel mechanistic insight into CEACAM3-initiated phagocytosis. We suggest that the CEACAM3 ITAM-like sequence is optimized to co-ordinate a minimal set of cellular factors needed to efficiently trigger actin-based lamellipodial protrusions and rapid pathogen engulfment

Instances and connectors : issues for a second generation process language

This work is supported by UK EPSRC grants GR/L34433 and GR/L32699Over the past decade a variety of process languages have been defined, used and evaluated. It is now possible to consider second generation languages based on this experience. Rather than develop a second generation wish list this position paper explores two issues: instances and connectors. Instances relate to the relationship between a process model as a description and the, possibly multiple, enacting instances which are created from it. Connectors refers to the issue of concurrency control and achieving a higher level of abstraction in how parts of a model interact. We believe that these issues are key to developing systems which can effectively support business processes, and that they have not received sufficient attention within the process modelling community. Through exploring these issues we also illustrate our approach to designing a second generation process language.Postprin

Building the New Europe: Western and Eastern Roads to Social Partnership

[Excerpt] While the ways in which neoliberalism and economic integration undermine social partnership and the welfare state have been extensively studied, less attention has been given to the ways in which such economic forces may push actors together, in reinvigorated bargaining relationships, to find workable solutions to difficult problems. In his article, we examine the contemporary status of social partnership in four case study countries—Germany, the United Kingdom, Bulgaria and Poland—as well as for Europe as a whole. In the west, while Germany presents a case of established social partnership under pressure, the United Kingdom has stood over the past two decades on the opposite neoliberal side. In the east, Bulgaria is one of the more developed cases of post-communist tripartism, while Poland exemplifies a weaker tripartism that emerged at a later stage of the transformation process. In selecting more and less developed social partnership cases in both west and east, we test the argument that the rise of Thatcher/Reagan/ Friedman ‘free market economics’ is paradoxically driving a resurgence and consolidation of social partnership relations across the new (both western and eastern) Europe