Subunit assembly modulates the activities of the Type III restriction–modification enzyme PstII in vitro

We demonstrate that, like other Type III restriction endonuclease, PstII does not turnover such that a DNA substrate is only fully cleaved at a Res(2)Mod(2)-to-site ratio of ∼1:1. However, unlike other Type III enzymes, the cleavage rate profiles varied with protein concentration: using 5 nM DNA and 25 nM PstII, approximately half of the DNA was cut at a fast rate while the remainder was cut 24 times more slowly; in comparison, with 100 nM PstII cleavage occurs at a single fast rate. The inclusion of the methyl donor S-adenosyl methionine does not alter the rates with 100 nM PstII but with 25 nM PstII the reaction stopped after completion of the initial fast cleavage phase owing to methylation. Concentration-dependent rates were also observed in methylation assays: at 100 nM PstII, a single slow rate was measured while at lower PstII concentrations both fast and slow rates were measured. We propose a model in which the intact Res(2)Mod(2) complex favoured at high PstII concentrations is a fast endonuclease/slow methyltransferase while the various subassemblies which coexist at lower concentrations are fast methyltransferases. A potential role for disassembly in control of restriction activity in vivo is discussed

Theoretical Investigation of Small Polyatomic Ions Observed in Inductively Coupled Plasma Mass Spectrometry: HxCO+ and HxN2+ (x = 1, 2, 3)

Two series of small polyatomic ions, HxCO+ and HxN2+ (x = 1, 2, 3), were systematically characterized using three correlated theoretical techniques:  density functional theory using the B3LYP functional, spin-restricted second-order perturbation theory, and singles + doubles coupled cluster theory with perturbative triples. On the basis of thermodynamic data, the existence of these ions in inductively coupled plasma mass spectrometry (ICP-MS) experiments is not surprising since the ions are predicted to be considerably more stable than their corresponding dissociation products (by 30−170 kcal/mol). While each pair of isoelectronic ions exhibit very similar thermodynamic and kinetic characteristics, there are significant differences within each series. While the mechanism for dissociation of the larger ions occurs through hydrogen abstraction, the triatomic ions (HCO+ and HN2+) appear to dissociate by proton abstraction. These differing mechanisms help to explain large differences in the abundances of HN2+ and HCO+ observed in ICP-MS experiments

What Is It All For? : The Intentions and Priorities for Study Abroad in Canadian Teacher Education

Within the research literature and in public discourse on higher education, attention has focused on the need for new graduates to develop 21st century skills for success in an increasingly globalized world. Calls for institutions of higher education to support student mobility abound, with intentions that some have categorized as neoliberal and others ascribe to notions of global citizenship. In this paper, we bring together literature from the fields of internationalization, teacher education, and study abroad to provide a conceptual framing and response to an inquiry into the following research question: In what ways does a study abroad experience support the development of preservice teachers? Through a multi-phase, multiple-perspective case study approach, we draw on qualitative interview data to illuminate how faculties of education and their students conceptualize the role of study abroad in the development of preservice teachers. The intentions for these programs cluster under four themes: global citizenship, personal growth, professional development, and employability. The concept of structured encounters with difference emerges out of these themes as a conceptual frame for future study abroad initiatives

Effects of Cry1Ab-Expressing Corn Anthers on Monarch Butterfly Larvae

Previous studies suggest that exposure to corn, Zea mays L., anthers expressing Bacillus thuringiensis (Bt)-derived protein may have adverse effects on the larvae of monarch butterfly, Danaus plexippus (L.). To examine the potential effects of Bt anthers on monarch butterflies, studies were designed to test toxicity in the laboratory; examine anther distribution in space and time; compare distributions of anthers, pollen, and larval feeding; and measure effects of long-term exposure in the field. In the laboratory, monarch butterfly larvae fed on whole corn anthers, but anther feeding was sporadic. Larvae exposed to 0.3 anther/cm2 fed and weighed less after 4 d compared with larvae exposed to non-Bt anthers. Adverse effects increased with increasing anther density. Monarch butterfly larvae exposed to 0.9 anther/cm2had reduced feeding, weight, and survival and increased developmental time compared with larvae exposed to non-Bt anthers. Later instars were more tolerant of Bt toxin. For all studies, laboratory testing probably magnified effects because larvae were confined to petri dishes. Field studies showed toxic anther densities are uncommon on milkweed (Asclepias) leaves in and near cornfields during anthesis. Mean anther densities on milkweed leaves in cornfields during peak anthesis were between 0.06 and 0.1 anther/cm2 (≈ 3–5 anthers per leaf). When exposure to a density of five anthers per leaf was tested in field-cage studies, no effects on growth, development, or survival were detected. Based on probability of exposure to toxic densities, Bt anthers alone are not likely to pose a significant risk to monarch butterflies in Iowa

Parallel tempering Monte Carlo in LAMMPS.

We present here the details of the implementation of the parallel tempering Monte Carlo technique into a LAMMPS, a heavily used massively parallel molecular dynamics code at Sandia. This technique allows for many replicas of a system to be run at different simulation temperatures. At various points in the simulation, configurations can be swapped between different temperature environments and then continued. This allows for large regions of energy space to be sampled very quickly, and allows for minimum energy configurations to emerge in very complex systems, such as large biomolecular systems. By including this algorithm into an existing code, we immediately gain all of the previous work that had been put into LAMMPS, and allow this technique to quickly be available to the entire Sandia and international LAMMPS community. Finally, we present an example of this code applied to folding a small protein

Fluctuating magnetic moments in liquid metals

We re-analyze literature data on neutron scattering by liquid metals to show that non-magnetic liquid metals possess a magnetic moment that fluctuates on a picosecond time scale. This time scale follows the motion of the cage-diffusion process in which an ion rattles around in the cage formed by its neighbors. We find that these fluctuating magnetic moments are present in liquid Hg, Al, Ga and Pb, and possibly also in the alkali metals.Comment: 17 pages, 5 figures, submitted to PR

Effects on Monarch Butterfly Larvae (Lepidoptera: Danaidae) After Continuous Exposure to Cry1Ab-Expressing Corn During Anthesis

Effects on monarch butterfly, Danaus plexippus L., after continuous exposure of larvae to natural deposits of Bacillus thuringiensis (Bt) and non-Bt pollen on milkweed, were measured in five studies. First instars were exposed at 3–4 and 6–7 d after initial anthesis, either directly on milkweed plants in commercial cornfields or in the laboratory on leaves collected from milkweeds in corn plots. Pollen exposure levels ranging from 122 to 188 grains/cm2/d were similar to within-field levels that monarch butterfly populations might experience in the general population of cornfields. Results indicate that 23.7% fewer larvae exposed to these levels of Bt pollen during anthesis reached the adult stage. A risk assessment procedure used previously was updated with a simulation model estimating the proportion of second-generation monarch butterflies affected. When considered over the entire range of the Corn Belt, which represents only 50% of the breeding population, the risk to monarch butterfly larvae associated with long-term exposure to Bt corn pollen is 0.6% additional mortality. Exposure also prolonged the developmental time of larvae by 1.8 d and reduced the weights of both pupae and adults by 5.5%. The sex ratio and wing length of adults were unaffected. The ecological significance of these sublethal effects is discussed relative to generation mortality and adult performance

Hidden Collections Redux: Summer 2013

This report is offers an update on the hidden collections at the UNT Libraries. Its purpose is to serve as a true representation of the collections in the UNT Libraries with little or no access

Pre-validation of a MALDI MS proteomics-based method for the reliable detection of blood and blood provenance

Abstract: The reliable identification of blood, as well as the determination of its origin (human or animal) is of great importance in a forensic investigation. Whilst presumptive tests are rapid and deployed in situ, their very nature requires confirmatory tests to be performed remotely. However, only serological tests can determine blood provenance. The present study improves on a previously devised Matrix Assisted Laser Desorption Ionisation Mass Spectrometry (MALDI MS)—proteomics based method for the reliable detection of blood by enabling the determination of blood provenance. The overall protocol was developed to be more specific than presumptive tests and faster/easier than the gold standard liquid chromatography (LC) MS/MS analysis. This is considered a pre-validation study that has investigated stains and fingermarks made in blood, other biofluids and substances that can elicit a false-positive response to colorimetric or presumptive tests, in a blind fashion. Stains and marks were either untreated or enhanced with a range of presumptive tests. Human and animal blood were correctly discriminated from other biofluids and non-biofluid related matrices; animal species determination was also possible within the system investigated. The procedure is compatible with the prior application of presumptive tests. The refined strategy resulting from iterative improvements through a trial and error study of 56 samples was applied to a final set of 13 blind samples. This final study yielded 12/13 correct identifications with the 13th sample being correctly identified as animal blood but with no species attribution. This body of work will contribute towards the validation of MALDI MS based methods and deployment in violent crimes involving bloodshed