Alternative catalytic residues in the active site of Esco acetyltransferases.

Cohesin is a protein complex whose core subunits, Smc1, Smc3, Scc1, and SA1/SA2 form a ring-like structure encircling the DNA. Cohesins play a key role in the expression, repair, and segregation of eukaryotic genomes. Following a catalytic mechanism that is insufficiently understood, Esco1 and Esco2 acetyltransferases acetylate the cohesin subunit Smc3, thereby inducing stabilization of cohesin on DNA. As a prerequisite for structure-guided investigation of enzymatic activity, we determine here the crystal structure of the mouse Esco2/CoA complex at 1.8 Å resolution. We reconstitute cohesin as tri- or tetrameric assemblies and use those as physiologically-relevant substrates for enzymatic assays in vitro. Furthermore, we employ cell-based complementation studies in mouse embryonic fibroblast deficient for Esco1 and Esco2, as a means to identify catalytically-important residues in vivo. These analyses demonstrate that D567/S566 and E491/S527, located on opposite sides of the murine Esco2 active site cleft, are critical for catalysis. Our experiments support a catalytic mechanism of acetylation where residues D567 and E491 are general bases that deprotonate the ε-amino group of lysine substrate, also involving two nearby serine residues - S566 and S527- that possess a proton relay function

Microchannel neural interface manufacture by stacking silicone and metal foil laminae

Objective: Microchannel neural interfaces (MNIs) overcome problems with recording from peripheral nerves by amplifying signals independent of Node of Ranvier position. Selective recording and stimulation using an MNI requires good insulation between microchannels and a high electrode density. We propose that stacking microchannel laminae will improve selectivity over single layer MNI designs due to the increase in electrode number and an improvement in microchannel sealing. Approach: This paper describes a manufacturing method for creating MNIs which overcomes limitations on electrode connectivity and microchannel sealing. Laser cut silicone – metal foil laminae were stacked using plasma bonding to create an array of microchannels containing tripolar electrodes. Electrodes were DC etched and electrode impedance and cyclic voltammetry were tested. Main results: MNIs with 100 µm and 200 µm diameter microchannels were manufactured. High electrode density MNIs are achievable with electrodes present in every microchannel. Electrode impedances of 27.2 ± 19.8 kΩ at 1kHz were achieved. Following 2 months of implantation in Lewis rat sciatic nerve, micro-fascicles were observed regenerating through the MNI microchannels. Significance: Selective microchannel neural interfaces with the peripheral nervous system may allow upper limb amputees to control prostheses intuitively

Drivers of grassland loss in Hungary during the post-socialist transformation (1987–1999)

The increase in the speed of land-cover change experienced worldwide is becoming a growing concern. Major socio-economic transitions, such as the breakdown of socialism in Europe, may lead to particularly high rates of landscape transformations. In this paper we examined the loss of semi-natural grasslands in Hungary between 1987 and 1999. We studied the relationship between 9 potential driving forces and the fate of grasslands using logistic GLMs. Grassland loss was found to be very high (1.31 % per year), which is far higher than either before or after this period. The most influential predictors of grassland loss were environmental and landscape characteristics (soil type, area of remnant grassland patches), and the socio-economic context (distance to paved road, and nearest settlement, human population density). Several processes and relationships can only be understood from a historical perspective (e.g. large extent of afforestation, strong decrease of soil water table). Grassland loss during the study period emerged as a consequence of survival strategies of individual farmers seeking adaptation to the changing environmental and socio-economic conditions, and not urbanization and agricultural intensification which are the main underlying drivers for the ongoing landscape transformations in most parts of the developed world. Though globalization increasingly influences local land use decisions , reconstructing and modelling recent landscape changes cannot be done without a proper understanding of local history and culture. Our analysis shows the importance of large-area yet high resolution landscape change research, which may reveal unexpected patterns of land cover change, undetected at coarser scales

Promoting communication skills for information systems students in Australian and Portuguese higher education : action research study

This paper aims to examine the value of communication skills learn- ing process through various assessments in Information Systems (IS) postgrad- uate units in Australia and Portugal. Currently, communication skills are indispensable to students in expanding their social networks and their knowl- edge at university and in the future workplace, since businesses expect their employees to have strong communication and presentation skills. This paper provides empirical evidence based on the anonymous quantitative and qualita- tive data collected during 2 years from 126 postgraduate students, which were collected via formal and informal feedback. Various assessment methods were used in Information Systems units to promote and develop the communication skills; these assessments are: reflective journal, business plan and prototype, discussion forum, presentation, and final examination. A Communication skills model (CSM) was developed based on Action research principles to promote the assessments which will assist IS students to enhance their communication skills. The research outcomes indicated that integrating communication skills in the assessments will allow students to promote their communication skills and boost their self-esteem skills. Furthermore, this paper added a new theoretical and practical contribution to higher-education teaching and learning literature, especially the action research for teachers to promote and develop communica- tion skills among students. Finally, integrating these skills in the units should meet the objectives and aims of the units, Master’s degrees, universities, and businesses’ needs, and satisfy our student’s need

Real-time three-dimensional transthoracic echocardiography in daily practice: initial experience

<p>Abstract</p> <p>Aim of the work</p> <p>To evaluate the feasibility and possible additional value of transthoracic real-time three-dimensional echocardiography (RT3D-TTE) for the assessment of cardiac structures as compared to 2D-TTE.</p> <p>Methods</p> <p>320 patients (mean age 45 ± 8.4 years, 75% males) underwent 2D-TTE and RT3D-TTE using 3DQ-Q lab software for offline analysis. Volume quantification and functional assessment was performed in 90 patients for left ventricle and in 20 patients for right ventricle. Assessment of native (112 patients) and prosthetic (30 patients) valves morphology and functions was performed. RT3D-TTE was performed for evaluation of septal defects in 30 patients and intracardiac masses in 52 patients.</p> <p>Results</p> <p>RT3D-TTE assessment of left ventricle was feasible and reproducible in 86% of patients while for right ventricle, it was (55%). RT3D-TTE could define the surface anatomy of mitral valve optimally (100%), while for aortic and tricuspid was (88% and 81% respectively). Valve area could be planimetered in 100% for the mitral and in 80% for the aortic. RT3D-TTE provided a comprehensive anatomical and functional evaluation of prosthetic valves. RT3D-TTE enface visualization of septal defects allowed optimal assessment of shape, size, area and number of defects and evaluated the outcome post device closure. RT3D-TTE allowed looking inside the intracardiac masses through multiple sectioning, valuable anatomical delineation and volume calculation.</p> <p>Conclusion</p> <p>Our initial experience showed that the use of RT3D-TTE in the assessment of cardiac patients is feasible and allowed detailed anatomical and functional assessment of many cardiac disorders.</p

Local Government Revenue Mobilisation in Anglophone Africa

This paper examines opportunities and constraints facing local revenue mobilisation in anglophone Africa, with an emphasis on urban settings. It discusses specific revenue instruments and their effects on economic efficiency, income distribution and accountability. In particular, it addresses political and administrative constraints facing various revenue instruments and factors affecting citizens’ compliance. The analysis is illustrated with examples from across anglophone Africa. A general conclusion emerging from the study is that local revenues mobilised in most local government authorities in Africa are necessary but not sufficient to develop and supply adequate services for the fast-growing population. On this basis, areas for further research on local government revenue mobilisation in Africa are identifiedDfI

Impairment of Immunoproteasome Function by β5i/LMP7 Subunit Deficiency Results in Severe Enterovirus Myocarditis

Proteasomes recognize and degrade poly-ubiquitinylated proteins. In infectious disease, cells activated by interferons (IFNs) express three unique catalytic subunits β1i/LMP2, β2i/MECL-1 and β5i/LMP7 forming an alternative proteasome isoform, the immunoproteasome (IP). The in vivo function of IPs in pathogen-induced inflammation is still a matter of controversy. IPs were mainly associated with MHC class I antigen processing. However, recent findings pointed to a more general function of IPs in response to cytokine stress. Here, we report on the role of IPs in acute coxsackievirus B3 (CVB3) myocarditis reflecting one of the most common viral disease entities among young people. Despite identical viral load in both control and IP-deficient mice, IP-deficiency was associated with severe acute heart muscle injury reflected by large foci of inflammatory lesions and severe myocardial tissue damage. Exacerbation of acute heart muscle injury in this host was ascribed to disequilibrium in protein homeostasis in viral heart disease as indicated by the detection of increased proteotoxic stress in cytokine-challenged cardiomyocytes and inflammatory cells from IP-deficient mice. In fact, due to IP-dependent removal of poly-ubiquitinylated protein aggregates in the injured myocardium IPs protected CVB3-challenged mice from oxidant-protein damage. Impaired NFκB activation in IP-deficient cardiomyocytes and inflammatory cells and proteotoxic stress in combination with severe inflammation in CVB3-challenged hearts from IP-deficient mice potentiated apoptotic cell death in this host, thus exacerbating acute tissue damage. Adoptive T cell transfer studies in IP-deficient mice are in agreement with data pointing towards an effective CD8 T cell immune. This study therefore demonstrates that IP formation primarily protects the target organ of CVB3 infection from excessive inflammatory tissue damage in a virus-induced proinflammatory cytokine milieu