Generation and characterization of monoclonal antibodies that recognize human and murine supervillin protein isoforms

Supervillin isoforms have been implicated in cell proliferation, actin filament-based motile processes, vesicle trafficking, and signal transduction. However, an understanding of the roles of these proteins in cancer metastasis and physiological processes has been limited by the difficulty of obtaining specific antibodies against these highly conserved membrane-associated proteins. To facilitate research into the biological functions of supervillin, monoclonal antibodies were generated against the bacterially expressed human supervillin N-terminus. Two chimeric monoclonal antibodies with rabbit Fc domains (clones 1E2/CPTC-SVIL-1; 4A8/CPTC-SVIL-2) and two mouse monoclonal antibodies (clones 5A8/CPTC-SVIL-3; 5G3/CPTC-SVIL-4) were characterized with respect to their binding sites, affinities, and for efficacy in immunoblotting, immunoprecipitation, immunofluorescence microscopy and immunohistochemical staining. Two antibodies (1E2, 5G3) recognize a sequence found only in primate supervillins, whereas the other two antibodies (4A8, 5A8) are specific for a more broadly conserved conformational epitope(s). All antibodies function in immunoblotting, immunoprecipitation and in immunofluorescence microscopy under the fixation conditions identified here. We also show that the 5A8 antibody works on immunohistological sections. These antibodies should provide useful tools for the study of mammalian supervillins

Possibilities for pedagogy in Further Education: Harnessing the abundance of literacy

In this report, it is argued that the most salient factor in the contemporary communicative landscape is the sheer abundance and diversity of possibilities for literacy, and that the extent and nature of students' communicative resources is a central issue in education. The text outlines the conceptual underpinnings of the Literacies for Learning in Further Education project in a social view of literacy, and the associated research design, methodology and analytical framework. It elaborates on the notion of the abundance of literacies in students' everyday lives, and on the potential for harnessing these as resources for the enhancement of learning. It provides case studies of changes in practice that have been undertaken by further education staff in order to draw upon students' everyday literacy practices on Travel and Tourism and Multimedia courses. It ends with some of the broad implications for conceptualising learning that arise from researching through the lens of literacy practices

Nature tourism and Irish film

This article provides a historical overview and reading of seminal Irish film from the perspective of nature tourism. Within Irish cultural studies, tourism is frequently equated with an overly romantic image of the island, which has been used to sell the country abroad. However, using notions like the tourist gaze and taking on board influential debates around space/place, one can posit a more progressive environmental vision of nature and landscape in our readings of film

Development and Deployment of a Bioreactor for the Removal of Sulfate and Manganese from Circumneutral Coal Mine Drainage

Surface mining, in the form of contour mining and mountain-top removal, is a common means for retrieving coal in the Appalachian Coal Belt region of Kentucky. Overburden or excess spoil generated by these two methods is placed in valley fills. Traditionally Constructed fills have been shown to adversely impact headwater ecosystems via stream burial and through alterations to the hydrology, sediment supply, water quality and biological composition of downstream reaches. Mine drainages emanating from the toe of valley fills often contain elevated levels of total dissolved solids and heavy metals. Drainage chemistry from Guy Cove, a valley fill located in eastern Kentucky, exhibited a mean pH of 6.5 and Fe, Mn and SO4 concentrations of 1.5, 14, and 1264 mg L-1, respectively. The objective of this research was to develop an anaerobic bioreactor for the purpose of reducing Mn and SO4 concentrations in the mine drainage. Development began with batch experiments that tested five different organic carbon sources and five different inorganic substrates. A synthetic mine drainage with a pH of 6.2 and Mn and SO4 concentrations of 90 and 1,500 mg/L, respectively, was used in the experiment. Manganese and SO4 removal varied widely between treatment matrices, with removal rates \u3c 10 to 100% for Mn and \u3c 10 to \u3e 80% for SO4. The substrate sources which provided the most treatment were hardwood mulch and biosolids combined with creek sediment. Subsequent experiments were performed using the synthetic mine drainage in small bioreactors (55 liter plastic tanks) filled with creek sediment with either hardwood mulch or biosolids, each replicated three times. Over a 65 day treatment period \u3e 90% of the Mn and 70% of the SO4 was removed. There were no statistical differences between the two organic substrates. Using this information, in-situ bioreactors consisting of two 5,500 liter plastic septic tanks filled with creek sediment, hardwood mulch and manure compost were installed at Guy Cove. Mine drainage was collected in a sump and conveyed through the inline bioreactors by gravity. Gate valves were used to control flow through the bioreactors. After a 10-month monitoring period, the in-situ bioreactors removed 12, 11, and 64% of Mn, SO4 and Fe from the drainage, respectively. Results from the field differed greatly from those observed under controlled laboratory conditions. Efforts to improve the efficiency of the in-situ bioreactors are underway

Recombination rate and selection strength in HIV intra-patient evolution

The evolutionary dynamics of HIV during the chronic phase of infection is driven by the host immune response and by selective pressures exerted through drug treatment. To understand and model the evolution of HIV quantitatively, the parameters governing genetic diversification and the strength of selection need to be known. While mutation rates can be measured in single replication cycles, the relevant effective recombination rate depends on the probability of coinfection of a cell with more than one virus and can only be inferred from population data. However, most population genetic estimators for recombination rates assume absence of selection and are hence of limited applicability to HIV, since positive and purifying selection are important in HIV evolution. Here, we estimate the rate of recombination and the distribution of selection coefficients from time-resolved sequence data tracking the evolution of HIV within single patients. By examining temporal changes in the genetic composition of the population, we estimate the effective recombination to be r=1.4e-5 recombinations per site and generation. Furthermore, we provide evidence that selection coefficients of at least 15% of the observed non-synonymous polymorphisms exceed 0.8% per generation. These results provide a basis for a more detailed understanding of the evolution of HIV. A particularly interesting case is evolution in response to drug treatment, where recombination can facilitate the rapid acquisition of multiple resistance mutations. With the methods developed here, more precise and more detailed studies will be possible, as soon as data with higher time resolution and greater sample sizes is available.Comment: to appear in PLoS Computational Biolog

Pair-instability and super-luminous supernova discoveries at z = 2.05, z = 2.50, and z = 3.90

We present the discovery of three super-luminous supernovae (SLSNe) at z = 2 - 4 as part of our survey to detect ultraviolet-luminous supernova at z > 2. SLSNe are ≥10 times more luminous than normal supernova types, reaching peak luminosities of ≳10^(44) erg s^(−1). A small subset of SLSNe (type SLSN-R) exhibit a slow evolution, and thus enormous integrated energies (≳10^(51) erg), consistent with the radiative decay of several solar masses of 56 Ni. SLSN-R are believed to be the deaths of very massive stars, ∼140 - 260 M_⊙, that are theorized to result in pair-instability supernovae. Two of the high redshift SLSNe presented here are consistent with the behavior of SLSN-R out to the extent in which their light curves are sampled, with the third event being consistent with the more rapid fade of the type II-L SLSN SN 2008es at z = 0.205. SLSNe are extremely rare locally but are expected to have been more common in the early Universe and as members of the first generation of stars to form after the Big Bang, the Population III stars. The high intrinsic luminosity of SLSNe and their detectability using our image-stacking technique out to z ∼ 6 provide the first viable route to detect and study the deaths of massive Population III stars which are expected to form in pristine gas at redshifts as low as z ∼ 2

The literacy practices for assessment in the vocational curriculum - the case of Hospitality

This article explores two case studies of the literacy practices of assessment in the vocational curriculum. Previous studies have identified learning in the vocational curriculum as being assessment-driven and that subjects often associated with limited levels of literacy actually require multiple literacy practices from students. Drawing upon studies of the literacy practices of assessment in the teaching of Hospitality, the study explores the academic and occupational forms of reading and writing required of students. These literacy practices are often invisible to both students and staff and yet are central to the success of students in relation to attainment. The article suggests that vocational subjects may actually be more demanding in the range of literacies required for assessment than is often considered to be the case

Individualized decision aid for diverse women with lupus nephritis (IDEA-WON): A randomized controlled trial.

BackgroundTreatment decision-making regarding immunosuppressive therapy is challenging for individuals with lupus. We assessed the effectiveness of a decision aid for immunosuppressive therapy in lupus nephritis.Methods and findingsIn a United States multicenter, open-label, randomized controlled trial (RCT), adult women with lupus nephritis, mostly from racial/ethnic minority backgrounds with low socioeconomic status (SES), seen in in- or outpatient settings, were randomized to an individualized, culturally tailored, computerized decision aid versus American College of Rheumatology (ACR) lupus pamphlet (1:1 ratio), using computer-generated randomization. We hypothesized that the co-primary outcomes of decisional conflict and informed choice regarding immunosuppressive medications would improve more in the decision aid group. Of 301 randomized women, 298 were analyzed; 47% were African-American, 26% Hispanic, and 15% white. Mean age (standard deviation [SD]) was 37 (12) years, 57% had annual income of <$40,000, and 36% had a high school education or less. Compared with the provision of the ACR lupus pamphlet (n = 147), participants randomized to the decision aid (n = 151) had (1) a clinically meaningful and statistically significant reduction in decisional conflict, 21.8 (standard error [SE], 2.5) versus 12.7 (SE, 2.0; p = 0.005) and (2) no difference in informed choice in the main analysis, 41% versus 31% (p = 0.08), but clinically meaningful and statistically significant difference in sensitivity analysis (net values for immunosuppressives positive [in favor] versus negative [against]), 50% versus 35% (p = 0.006). Unresolved decisional conflict was lower in the decision aid versus pamphlet groups, 22% versus 44% (p < 0.001). Significantly more patients in the decision aid versus pamphlet group rated information to be excellent for understanding lupus nephritis (49% versus 33%), risk factors (43% versus 27%), medication options (50% versus 33%; p ≤ 0.003 for all); and the ease of use of materials was higher in the decision aid versus pamphlet groups (51% versus 38%; p = 0.006). Key study limitations were the exclusion of men, short follow-up, and the lack of clinical outcomes, including medication adherence.ConclusionsAn individualized decision aid was more effective than usual care in reducing decisional conflict for choice of immunosuppressive medications in women with lupus nephritis.Trial registrationClinicaltrials.gov, NCT02319525

Present-day stress orientations and tectonic provinces of the NW Borneo collisional margin

Extent: 15p.Borehole failure observed on image and dipmeter logs from 55 petroleum wells across the NW Borneo collisional margin were used to determine maximum horizontal stress (σH) orientations; combined with seismic and outcrop data, they define seven tectonic provinces. The Baram Delta–Deepwater Fold-Thrust Belt exhibits three tectonic provinces: its inner shelf inverted province (σH is NW-SE, margin-normal), its outer shelf extension province (σH is NE-SW, margin-parallel), and its slope to basin floor compression province (σH is NW-SE, margin-normal). In the inverted province, σH reflects inversion of deltaic normal faults. The σH orientations in the extension and compression provinces reflect deltaic gravitational tectonics. The shale and minibasin provinces have been recognized in offshore Sabah. In the shale province, σH is N010°E, which aligns around the boundary of a massif of mobile shale. Currently, no data are available to determine σH in the minibasin province. In the Balingian province, σH is ESE-WNW, reflecting ESE absolute Sunda plate motions due to the absence of a thick detachment seen elsewhere in NW Borneo. The Central Luconia province demonstrates poorly constrained and variable σH orientations. These seven provinces result from the heterogeneous structural and stratigraphic development of the NW Borneo margin and formed due to complex collisional tectonics and the varied distribution and thicknesses of stratigraphic packages.Rosalind C. King, Mark R. P. Tingay, Richard R. Hillis, Christopher K. Morley, and James Clar

Open science in archaeology

No abstract available