The degradation of p53 and its major E3 ligase Mdm2 is differentially dependent on the proteasomal ubiquitin receptor S5a.

p53 and its major E3 ligase Mdm2 are both ubiquitinated and targeted to the proteasome for degradation. Despite the importance of this in regulating the p53 pathway, little is known about the mechanisms of proteasomal recognition of ubiquitinated p53 and Mdm2. In this study, we show that knockdown of the proteasomal ubiquitin receptor S5a/PSMD4/Rpn10 inhibits p53 protein degradation and results in the accumulation of ubiquitinated p53. Overexpression of a dominant-negative deletion of S5a lacking its ubiquitin-interacting motifs (UIM)s, but which can be incorporated into the proteasome, also causes the stabilization of p53. Furthermore, small-interferring RNA (siRNA) rescue experiments confirm that the UIMs of S5a are required for the maintenance of low p53 levels. These observations indicate that S5a participates in the recognition of ubiquitinated p53 by the proteasome. In contrast, targeting S5a has no effect on the rate of degradation of Mdm2, indicating that proteasomal recognition of Mdm2 can be mediated by an S5a-independent pathway. S5a knockdown results in an increase in the transcriptional activity of p53. The selective stabilization of p53 and not Mdm2 provides a mechanism for p53 activation. Depletion of S5a causes a p53-dependent decrease in cell proliferation, demonstrating that p53 can have a dominant role in the response to targeting S5a. This study provides evidence for alternative pathways of proteasomal recognition of p53 and Mdm2. Differences in recognition by the proteasome could provide a means to modulate the relative stability of p53 and Mdm2 in response to cellular signals. In addition, they could be exploited for p53-activating therapies. This work shows that the degradation of proteins by the proteasome can be selectively dependent on S5a in human cells, and that this selectivity can extend to an E3 ubiquitin ligase and its substrate

Dependent self-employment: workers between employment and self-employment in the UK

Analysing the British Labour Force Survey, we highlight that dependent self-employed workers constitute a group distinct from both employees and independent self-employed workers in the labour market group. Dependent self-employed workers show characteristics of a more volatile labour market attachment than employed or self-employed workers. We provide empirical evidence that dependent self-employed workers are rather pushed than pulled into this labour market status, making dependent self-employment an example of 'necessity' rather than 'opportunity' entrepreneurship. Although data limitations only allow a limited longitudinal analysis, we provide evidence that the majority of dependent self-employed workers remain in the labour market in the short run - either as self-employed or employed - and that only few leave the labour market. In addition, dependent self-employment does not create jobs for others; in our data, dependent self-employed individuals stop being dependent and self-employed because they increase their customer base or return to paid employment

Reassessing the Impact of High Performance Workplaces

High performance workplace practices were extolled as an efficient means to increase firm productivity. The empirical evidence is disputed, however. To assess the productivity effects of a broad variety of measures, we simultaneously account for both unobserved heterogeneity and endogeneity using establishment panel data for Germany. We show that increasing employee participation enhances firm productivity in Germany, whereas incentive systems do not foster productivity. Our results further indicate that firms with structural productivity problems tend to introduce organisational changes that increase employee participation whereas well performing firms are more likely to offer incentives

“Nonsense Rides Piggyback on Sensible Things”: The Past, Present, and Future of Graphology

“Nonsense rides piggyback on sensible things”, declares professional sceptic and questioned-document analyst Joe Nickell concerning graphology. This chapter examines graphology’s enduring allure and reach, despite its controversies, and considers its relationship with other types of handwriting analysis. It first asks: is it possible to metaphorically “dissect” the page of handwritten texts, to scrutinize writing as a “medical paratext” rich in information about the writer’s state of health? It then interrogates the nature of the connection between physical and mental states and handwriting. It demonstrates how academics are going “back to basics” with their enquiries into individual difference and handwriting features, and how digital methodologies are contributing to this. Thus, this chapter is an updated study of graphology, providing a wider understanding of the concept of the paratext by considering the information captured in handwriting in the context of a digital age

Ignicoccus hospitalis and Nanoarchaeum equitans: ultrastructure, cell–cell interaction, and 3D reconstruction from serial sections of freeze-substituted cells and by electron cryotomography

Ultrastructure and intercellular interaction of Ignicoccus hospitalis and Nanoarchaeum equitans were investigated using two different electron microscopy approaches, by three-dimensional reconstructions from serial sections, and by electron cryotomography. Serial sections were assembled into 3D reconstructions, for visualizing the unusual complexity of I. hospitalis, its huge periplasmic space, the vesiculating cytoplasmic membrane, and the outer membrane. The cytoplasm contains fibres which are reminiscent to a cytoskeleton. Cell division in I. hospitalis is complex, and different to that in Euryarchaeota or Bacteria. An irregular invagination of the cytoplasmic membrane is followed by separation of the two cytoplasms. Simultaneous constriction of cytoplasmic plus outer membrane is not observed. Cells of N. equitans show a classical mode of cell division, by constriction in the mid-plane. Their cytoplasm exhibits two types of fibres, elongated and ring-shaped. Electron micrographs of contact sites between I. hospitalis and N. equitans exhibit two modes of interaction. One is indirect and mediated by thin fibres; in other cells the two cell surfaces are in direct contact. The two membranes of I. hospitalis cells are frequently seen in direct contact, possibly a prerequisite for transporting metabolites or substrates from the cytoplasm of one cell to the other. Rarely, a transport based on cargo vesicles is observed between I. hospitalis and N. equitans

Isolation and Culture of Larval Cells from C. elegans

Cell culture is an essential tool to study cell function. In C. elegans the ability to isolate and culture cells has been limited to embryonically derived cells. However, cells or blastomeres isolated from mixed stage embryos terminally differentiate within 24 hours of culture, thus precluding post-embryonic stage cell culture. We have developed an efficient and technically simple method for large-scale isolation and primary culture of larval-stage cells. We have optimized the treatment to maximize cell number and minimize cell death for each of the four larval stages. We obtained up to 7.8×104 cells per microliter of packed larvae, and up to 97% of adherent cells isolated by this method were viable for at least 16 hours. Cultured larval cells showed stage-specific increases in both cell size and multinuclearity and expressed lineage- and cell type-specific reporters. The majority (81%) of larval cells isolated by our method were muscle cells that exhibited stage-specific phenotypes. L1 muscle cells developed 1 to 2 wide cytoplasmic processes, while L4 muscle cells developed 4 to 14 processes of various thicknesses. L4 muscle cells developed bands of myosin heavy chain A thick filaments at the cell center and spontaneously contracted ex vivo. Neurons constituted less than 10% of the isolated cells and the majority of neurons developed one or more long, microtubule-rich protrusions that terminated in actin-rich growth cones. In addition to cells such as muscle and neuron that are high abundance in vivo, we were also able to isolate M-lineage cells that constitute less than 0.2% of cells in vivo. Our novel method of cell isolation extends C. elegans cell culture to larval developmental stages, and allows use of the wealth of cell culture tools, such as cell sorting, electrophysiology, co-culture, and high-resolution imaging of subcellular dynamics, in investigation of post-embryonic development and physiology

Determinants of Capital Structure in Non-Financial Companies

In this paper, we evaluate firm-, industry- and country-specific factors determining a firm’s capital structure. The empirical validity of several capital structure theories has been ambiguous so far. We shed light on the main drivers of leverage and depict differences in industry and country characteristics. Using a short panel data set with a large cross-section, we are able to show that firm size, industry leverage, industry growth and tax shield positively affect leverage ratios, while profitability and liquidity have negative impacts. Moreover, our model is an improvement over Rajan and Zingales’ (1995) four-factor core model in terms of explaining data variation. The results are robust against different panel estimators, decompositions and over time

How are antibodies involved in the protective mechanism of susceptible mice infected with T. cruzi?

Host resistance to Trypanosoma cruzi is dependent on both natural and acquired immune responses. During the acute phase of the infection the presence of IFN-g, TNF-a, IL-12 and GM-CSF has been closely associated with resistance, whereas TGF-ß and IL-10 have been associated with susceptibility. Several investigators have demonstrated that antibodies are responsible for the survival of susceptible animals in the initial phase of infection and for the maintenance of low levels of parasitemia in the chronic phase. However, how this occurs is not yet understood. Our results and other data in the literature support the hypothesis that the protective role of antibodies in the acute phase of infection is dependent mostly on their ability to induce removal of bloodstream trypomastigotes from the circulation in addition to other concomitant cell-mediated events

Current Account Balances and Structural Adjustment in the Euro Area

In the past decade, a set of euro area countries has accumulated large current account deficits. After a brief relaxation of the euro area internal imbalances in the wake of the financial crisis, it appears as if this pattern arises anew when times normalize again and Germany still sticks to export-led growth. This issue has been labelled one of the most challenging economic policy issues for Europe inter alia by the European Commission and some other players on the EU level. In this paper, we analyse the role of private restructuring and structural reforms for the urgently needed sustainable readjustment of intra-euro area current account balances. A panel regression reveals a significant impact of structural reforms on intra-euro area current account balances. This implies that in particular structural reforms and wage restraint in notorious current account and budget deficit countries such as Greece are highly suitable to support long-term economic stability in Europe.In den vergangenen Jahren haben einige Staaten der EWU, besonders Griechenland, Spanien und Portugal, hohe Leistungsbilanzdefizite aufgebaut. Die Finanzmarktkrise führte zwar zu einer Entspannung, jedoch ist eine deutliche Reduzierung der Defizite nicht in Sicht. Insbesondere durch die Griechenland-Krise haben die Leistungsbilanzdefizite einen hohen Stellenwert auf der wirtschaftspolitischen Agenda der EU eingenommen. Das Papier untersucht, inwieweit private Restrukturierungsmaßnahmen der Privatwirtschaft und/oder Strukturreformen zu einer notwendigen und nachhaltigen Korrektur der Leistungsbilanzdefizite in der EWU beitragen können. Eine umfassende empirische Analyse zeigt die herausragende Bedeutung von Strukturreformen für die Anpassung der Leistungsbilanzen innerhalb der EWU. Der Effekt von Arbeitsmarktreformen ist besonders ausgeprägt. Dies unterstützt die Forderung nach umfassenden und nachhaltigen Strukturreformen in der EWU, besonders in den Ländern mit hohen Leistungsbilanzdefiziten. Dies würde die derzeitigen ökonomischen Spannungen in der EWU reduzieren und die Stabilität der EWU sowie des Euros erhöhen