Development of FRET-Based Assays in the Far-Red Using CdTe Quantum Dots

Colloidal quantum dots (QDs) are now commercially available in a biofunctionalized form, and Förster resonance energy transfer (FRET) between bioconjugated dots and fluorophores within the visible range has been observed. We are particularly interested in the far-red region, as from a biological perspective there are benefits in pushing to ∼700 nm to minimize optical absorption (ABS) within tissue and to avoid cell autofluorescence. We report on FRET between streptavidin- (STV-) conjugated CdTe quantum dots, Qdot705-STV, with biotinylated DY731-Bio fluorophores in a donor-acceptor assay. We also highlight the changes in DY731-Bio absorptivity during the streptavidin-biotin binding process which can be attributed to the structural reorientation. For fluorescence beyond 700 nm, different alloy compositions are required for the QD core and these changes directly affect the fluorescence decay dynamics producing a marked biexponential decay with a long-lifetime component in excess of 100 nanoseconds. We compare the influence of the two QD relaxation routes upon FRET dynamics in the presence of DY731-Bio

Mountain lions prey selectively on prion-infected mule deer

The possibility that predators choose prey selectively based on age or condition has been suggested but rarely tested. We examined whether mountain lions (Puma concolor) selectively prey upon mule deer (Odocoileus hemionus) infected with chronic wasting disease, a prion disease. We located kill sites of mountain lions in the northern Front Range of Colorado, USA, and compared disease prevalence among lion-killed adult (≥2 years old) deer with prevalence among sympatric deer taken by hunters in the vicinity of kill sites. Hunter-killed female deer were less likely to be infected than males (odds ratios (OR) = 0.2, 95% confidence intervals (CI) = 0.1–0.6; p = 0.015). However, both female (OR = 8.5, 95% CI = 2.3–30.9) and male deer (OR = 3.2, 95% CI = 1–10) killed by a mountain lion were more likely to be infected than same-sex deer killed in the vicinity by a hunter (p < 0.001), suggesting that mountain lions in this area actively selected prion-infected individuals when targeting adult mule deer as prey items

Enhancing the immunogenicity of tumour lysate-loaded dendritic cell vaccines by conjugation to virus-like particles

BACKGROUND: Tumour cell lysates are an excellent source of many defined and undefined tumour antigens and have been used clinically in immunotherapeutic regimes but with limited success. METHODS: We conjugated Mel888 melanoma lysates to rabbit haemorrhagic disease virus virus-like particles (VLP), which can act as vehicles to deliver multiple tumour epitopes to dendritic cells (DC) to effectively activate antitumour responses. RESULTS: Virus-like particles did not stimulate the phenotypic maturation of DC although, the conjugation of lysates to VLP (VLP-lysate) did overcome lysate-induced suppression of DC activation. Lysate-conjugated VLP enhanced delivery of antigenic proteins to DC, while the co-delivery of VLP-lysates with OK432 resulted in cross-priming of naïve T cells, with expansion of a MART1(+) population of CD8(+) T cells and generation of a specific cytotoxic response against Mel888 tumour cell targets. The responses generated with VLP-lysate and OK432 were superior to those stimulated by unconjugated lysate with OK432. CONCLUSION: Collectively, these results show that the combination of VLP-lysate with OK432 delivered to DC overcomes the suppressive effects of lysates, and enables priming of naïve T cells with superior ability to specifically kill their target tumour cells

Scenario-based eLearning to promote active learning in large cohorts: students’ perspective

This work presents results from an investigation on the students' perspective of the use of scenario‐based (SBL) e‐learning and their performance in a first‐year core chemical engineering module in a Scottish university. SBL is a pedagogy that promotes active learning by bringing to the classroom practical and industrial experience. When combined with online delivery, SBL can be used to increase students' engagement in large cohorts. The scenario developed and used in this work was delivered via the Scenario‐Based Learning Interactive tool. A survey was used to capture students' insights on the activity and the use of the software. These data were then analyzed in combination with students' exam performance. Two cohorts with around 100 students each participated in this study over 2 years (with 91% response rate in the evaluation questionnaire). They indicate that students fully engaged with this form of learning as it links module content with real industrial applications. There is, however, a significant difference between female and male students in terms of the enjoyment they derived from the computer‐based activity with male students preferring the activity over tutorials with opposite results for females. There is no relationship between the perceived level of difficulty of the scenario and the exam performance in either cohort. The majority of students identified that they developed their problem‐solving and analytical skills through doing the scenario activity. In general, the students found the software difficult to use which suggests the need to explore other tools for the delivery of scenario‐based activities

A large-scale experiment to evaluate control of invasive muskrats

The muskrat (Ondatra zibethicus) is an invasive species in Europe. The extensive waterways of the Netherlands provide ideal habitat for muskrats, and a large population established itself after arrival in 1941. A control program was put into effect immediately because muskrat burrowing can compromise the integrity of dikes and, hence, poses a significant public safety risk. The current (2015) annual catch of approximately 89,000 individuals is equivalent to approximately 0.30 muskrats/km of waterway, well above the national objective in spite of decades of effort. The control program is expensive (€35 M annually) and contested by animal rights groups. These factors created the need for a careful evaluation of the full range of control possibilities, from ‘no control’ to ‘extermination.’ As part of this, we experimentally evaluated the validity of a previously published correlation (based on historical data) between catch and effort. We raised or lowered removal effort (2013–2016) in a stratified random sample of 117 5-km × 5-km ‘atlas squares’ from the national grid. We found that catch-per-unit effort (CPUE) decreased after effort was increased, and rose after effort was decreased, by amounts slightly greater than expected based on the correlational data, though confidence intervals enclose zero. As anticipated, CPUE varied consistently and strongly between seasons. The biggest (and unanticipated) effects were those of the catch in the preceding 3 years (‘history’), and surrounding area (‘neighborhood’). Our experiment confirms estimates of intensity of control required to lower muskrat populations. These results will help with more effective allocation of control effort, and better-informed evaluation of the economic costs of various control options

Transport properties of dense fluid argon

We calculate using molecular dynamics simulations the transport properties of realistically modeled fluid argon at pressures up to $\simeq 50GPa$ and temperatures up to $3000K$. In this context we provide a critique of some newer theoretical predictions for the diffusion coefficients of liquids and a discussion of the Enskog theory relevance under two different adaptations: modified Enskog theory (MET) and effective diameter Enskog theory. We also analyze a number of experimental data for the thermal conductivity of monoatomic and small diatomic dense fluids.Comment: 8 pages, 6 figure

Ab initio van der Waals interactions in simulations of water alter structure from mainly tetrahedral to high-density-like

The structure of liquid water at ambient conditions is studied in ab initio molecular dynamics simulations using van der Waals (vdW) density-functional theory, i.e. using the new exchange-correlation functionals optPBE-vdW and vdW-DF2. Inclusion of the more isotropic vdW interactions counteracts highly directional hydrogen-bonds, which are enhanced by standard functionals. This brings about a softening of the microscopic structure of water, as seen from the broadening of angular distribution functions and, in particular, from the much lower and broader first peak in the oxygen-oxygen pair-correlation function (PCF), indicating loss of structure in the outer solvation shells. In combination with softer non-local correlation terms, as in the new parameterization of vdW-DF, inclusion of vdW interactions is shown to shift the balance of resulting structures from open tetrahedral to more close-packed. The resulting O-O PCF shows some resemblance with experiment for high-density water (A. K. Soper and M. A. Ricci, Phys. Rev. Lett., 84:2881, 2000), but not directly with experiment for ambient water. However, an O-O PCF consisting of a linear combination of 70% from vdW-DF2 and 30% from experiment on low-density liquid water reproduces near-quantitatively the experimental O-O PCF for ambient water, indicating consistency with a two-liquid model with fluctuations between high- and low-density regions

Expression of divIB of Bacillus subtilis during vegetative growth

Expression of the division initiation gene, divIB, of Bacillus subtilis vegetative growth was examined. lacZ fusion studies and transcription start point mapping have established that a sigma A promoter proximal to divIB is utilized in vivo. The -10 region of this promoter, which is located 93 bp upstream of the start codon, has been defined precisely by site-directed mutagenesis that destroys the promoter. Examination of transcripts by Northern (RNA) blotting has shown that there are at least two transcripts for divIB. The established proximal promoter was found to give rise to a very minor transcript which could not be convincingly demonstrated in wild-type cells but which became apparent upon insertion of a plasmid into the chromosome just upstream of this promoter. The major transcript for divIB originated from a site several kb upstream of the gene and is probably the same as the long polycistronic message also traversing the murD-spoVE-murG genes that was identified previously by others (A.D. Henriques, H. de Lencastre, and P.J. Piggot, Biochimie 74:735-748, 1992). Transcription from the proximal promoter alone, in an upstream-deletion mutant strain, provided sufficient DivIB for normal growth and division as well as sporulation