Dual-color bioluminescent bioreporter for forensic analysis: evidence of androgenic and anti-androgenic activity of illicit drugs

Bioassays represent promising complementary techniques to conventional analytical approaches used in doping analysis to detect illicit drugs like anabolic-androgenic steroids (AAS). The fact that all AAS share a common mechanism of action via the human androgen receptor (hAR) enables the use of bioassays, relying on the activation of hAR as antidoping screening tools. Previously, we developed a dual-color bioreporter based on yeast cells engineered to express hAR and androgen response elements driving the expression of the bioluminescent (BL) reporter protein Photinus pyralis luciferase. A second reporter protein, the red-emitting luciferase PpyRE8, was introduced in the bioreporter as internal viability control. Here, we report the first forensic application of a straightforward, accurate, and cost-effective bioassay, relying on spectral resolution of the two BL signals, in 96-microwell format. The bioreporter responds to dihydrotestosterone as reference androgen in a concentration-dependent manner from 0.08 to 1,000 nM with intra- and inter-assay variation coefficients of 11.4 % and 13.1 %, respectively. We also demonstrated the suitability of this dual-color bioreporter to assess (anti)-androgenic activity of pure AAS, mixtures of AAS, and other illicit drugs provided by the Scientific Police. Significant anti-androgenic activity was observed in samples labeled as marijuana and hashish, containing Δ⁹-tetrahydrocannabinol as major constituent.

Intermethod Comparison and Evaluation of Measured Near Surface Residual Stress in Milled Aluminum

Background

While near surface residual stress (NSRS) from milling is a driver for distortion in aluminum parts there are few studies that directly compare available techniques for NSRS measurement.

Objective

We report application and assessment of four different techniques for evaluating residual stress versus depth in milled aluminum parts.

Methods

The four techniques are: hole-drilling, slotting, cos(α) x-ray diffraction (XRD), and sin²(ψ) XRD, all including incremental material removal to produce a stress versus depth profile. The milled aluminum parts are cut from stress-relieved plate, AA7050-T7451, with a range of table and tool speeds used to mill a large flat surface in several samples. NSRS measurements are made at specified locations on each sample.

Results

Resulting data show that NSRS from three techniques are in general agreement: hole-drilling, slotting, and sin²(ψ) XRD. At shallow depths (<?0.03 mm), sin²(ψ) XRD data have the best repeatability (<?15 MPa), but at larger depths (>?0.04 mm) hole-drilling and slotting have the best repeatability (<?10 MPa). NSRS data from cos(α) XRD differ from data provided by other techniques and the data are less repeatable. NSRS data for different milling parameters show that the depth of NSRS increases with feed per tooth and is unaffected by cutting speed.

Conclusion

Hole-drilling, slotting, and sin²(ψ) XRD provided comparable results when assessing milling-induced near surface residual stress in aluminum. Combining a simple distortion test, comprising removal of a 1 mm thick wafer at the milled surface, with a companion stress analysis showed that NSRS data from hole-drilling are most consistent with milling-induced distortion.

     

Dynamics of communal vocalizations in a social songbird, the zebra finch (Taeniopygia guttata)

Colonies or communities of animals such as fishes, frogs, seabirds, or marine mammals can be noisy. Although vocal communication between clearly identified sender(s) and receiver(s) has been well studied, the properties of the noisy sound that results from the acoustic network of a colony of gregarious animals have received less attention. The resulting sound could nonetheless convey some information about the emitting group. Using custom-written software for automatic detection of vocalizations occurring over many hours of recordings, this study reports acoustic features of communal vocal activities in a gregarious species, the zebra finch (Taeniopygia guttata). By biasing the sex ratio and using two different housing conditions (individual versus communal housing), six groups of zebra finches were generated, with six different social structures that varied both in terms of sex-composition and proportion of paired individuals. The results showed that the rate of emission and the acoustic dynamic both depended on the social structure. In particular, the vocal activity of a group of zebra finches depended mainly on the number of unpaired birds, i.e., individuals not part of a stably bonded pair.     

A thermodynamic characterization of aqueous nanostructures of water molecules formed by prolonged contact with the hydrophilic polymer Nafion

This work investigates the physicochemical alterations of water perturbed by prolonged contact with the hydrophilic polymer Nafion, referred to as iteratively nafionized water (INW). The parameters measured are: electrical conductivity, χ μS cm^?1, heat of mixing with basic sodium hydroxide (NaOH) solutions, ΔQ _NaOH ^mix  J kg^?1, and pH. The results indicate that supramolecular aggregates of water molecules form after prolonged contact with a Nafion surface. Analytical determination by ion chromatography allows us to exclude the role of contaminants. This suggests that water may possess an exceptional self-organization capability triggered by the contact with a hydrophilic surface. Conductometric, pH-metric, and calorimetric titrations of INW were performed by the addition of NaOH solutions to determine the concentration of the aqueous nanostructures, via conductometric and pH-metric titration with NaOH solutions. Thermodynamic parameters were determined via calorimetric titration, for the process of formation of complexes between the nanostructures and the base used.     

Cell‐Permeant and Photocleavable Chemical Inducer of Dimerization

Chemical inducers of dimerization (CIDs) have been developed to orchestrate protein dimerization and translocation. Here we present a novel photocleavable HaloTag‐ and SNAP‐tag‐reactive CID (MeNV‐HaXS) with excellent selectivity and intracellular reactivity. Excitation at 360 nm cleaves the methyl‐6‐nitroveratryl core of MeNV‐HaXS. MeNV‐HaXS covalently links HaloTag‐ and SNAP‐tag fusion proteins, and enables targeting of selected membranes and intracellular organelles. MeNV‐HaXS‐mediated translocation has been validated for plasma membrane, late endosomes, lysosomes, Golgi, mitochondria, and the actin cytoskeleton. Photocleavage of MeNV‐HaXS liberates target proteins and provides access to optical manipulation of protein relocation with high spatiotemporal and subcellular precision. MeNV‐HaXS supports kinetic studies of protein dynamics and the manipulation of subcellular enzyme activities, which is exemplified for Golgi‐targeted cargo and the assessment of nuclear import kinetics.     

A remarkable accelerating effect of iodide ions in the photostimulated phosphonylation of bromoaromatic compounds

We find that 1 equiv. of NaI greatly accelerates the photostimulated substitution (S_RN¹ mechanism) of bromobenzene, bromotoluenes and bromopyridines by diethylphosphite ion, in acetonitrile/tetrahydrofuran mixtures. Consequently, bromoaromatic compounds are suitable for preparative use.     

Sur le groupe de la géométrie hypersphérique

Sans résumé     

Large-eddy simulation of transition to turbulence in a heated annular channel

One carries out three-dimensional large-eddy simulations of natural convection in a horizontal annulus using Smagorinsky's dynamic subgrid model. The onset of transition to turbulence and turbulent regimes are analyzed. The characteristics of unstable flows and their influence on the heat-transfer process are studied. To cite this article: E.L.M. Padilla, A. Silveira-Neto, C. R. Mecanique 333 (2005).     

Mathematical Working Spaces in schooling: an introduction

ZDM – Mathematics Education - The theoretical and methodological model of Mathematical Working Space (MWS) is introduced in this paper. For over 10&nbsp;years, the model has been the...