Urine testing for designer steroids by liquid chromatography with androgen bioassay detection and electrospray quadrupole time-of-flight mass spectrometry identification

New anabolic steroids show up occasionally in sports doping and in veterinary control. The discovery of these designer steroids is facilitated by findings of illicit preparations, thus allowing bioactivity testing, structure elucidation using NMR and mass spectrometry, and final incorporation in urine testing. However, as long as these preparations remain undiscovered, new designer steroids are not screened for in routine sports doping or veterinary control urine tests since the established GC/MS and LC/MS/MS methods are set up for the monitoring of a few selected ions or MS/MS transitions of known substances only. In this study, the feasibility of androgen bioactivity testing and mass spectrometric identification is being investigated for trace analysis of designer steroids in urine. Following enzymatic deconjugation and a generic solid-phase extraction, the samples are analyzed by gradient LC with effluent splitting toward two identical 96-well fraction collectors. One well plate is used for androgen bioactivity detection using a novel robust yeast reporter gene bioassay yielding a biogram featuring a 20-s time resolution. The bioactive wells direct the identification efforts to the corresponding well numbers in the duplicate plate. These are subjected to high-resolution LC using a short column packed with 1.7-microm C18 material and coupled with electrospray quadrupole time-of-flight mass spectrometry (LC/QTOFMS) with accurate mass measurement. Element compositions are calculated and used to interrogate electronic substance databases. The feasibility of this approach for doping control is demonstrated via the screening of human urine samples spiked with the designer anabolic steroid tetrahydrogestrinone. Application of the proposed methodology, complementary to the established targeted urine screening for known anabolics, will increase the chance of finding unknown emerging designer steroids, rather then being solely dependent on findings of the illicit preparations themselves.     

Humidity-Regulating Trays: Moisture Absorption Kinetics and Applications for Fresh Produce Packaging

For packaged fresh produce, inappropriate high relative humidity (RH) levels and condensation of water vapour cause premature spoilage. Humidity-regulating trays were developed to solve this issue. They were made from a thermoformed multilayer structure: polyethylene (outside)/foamed hygroscopic ionomer (active layer) with 0 or 12 wt% NaCl/hygroscopic ionomer (sealing layer, inside). Moisture absorption kinetics of the humidity-regulating trays with 0 and 12 wt% NaCl (T-0 and T-12, respectively) was investigated under different RH conditions (76, 86, 96 and 100 %) at 13 °C for 16 days. Additional trays containing 7 g of distilled water were closed with a high barrier lidding film, and the headspace RH was continuously monitored as a function of time. As control, a polypropylene (control-PP) tray was used. Strawberries and tomatoes were used to test capability of the trays to regulate in-package RH. The amount of water absorbed by the T-0 and T-12 trays was 7.6 and 13.2 g, respectively. Active hygroscopic ionomer layer was effective in water vapour absorption, and the integration of NaCl into this active layer increased the water vapour absorption capacity of the tray. The Weibull model adequately described the moisture sorption kinetics of the individual packaging trays as a function of time. The headspace RH of trays covered with a lidding film was found to be 89.8, 99.6 and 100 % in the T-12, T-0 and control-PP trays, respectively. The T-12 trays containing fresh produce best regulated the in-package RH below 97 % and maintained overall quality, but at the expense of slightly higher product weight loss (2–3 wt% for strawberry, 1 wt% for tomatoes) compared to the control-PP trays (0.3–0.6 wt%).     

Electron microprobe and synchrotron x-ray fluorescence mapping of the heterogeneous distribution of copper in high-copper vineyard soils

The response of microorganisms to metal contamination of soils varies significantly from one investigation to another. One explanation is that metals are heterogeneously distributed at spatial scales relevant to microbes and that microoorganisms are able to avoid zones of intense contamination. This article aims to assess the microscale distribution of Cu in a vineyard soil. The spatial distribution of Cu was measured at two resolutions (0.3 mm and 20 microm) in thin sections of the surface 4 cm of undisturbed soil by electron microprobe and synchrotron X-ray microfluo-rescence spectroscopy. Bulk physicochemical analyses of Cu, pH, organic matter, texture, and mineralogy were performed. The results indicate that the Cu distribution is strongly heterogeneous at both scales of observation. Entire regions of the thin sections are virtually devoid of Cu, whereas highly localized "hotspots" have Cu signal intensities thousands of times higher than background. The distribution of Rb, or Al and Si, indicators of clay minerals, or Fe (iron (hydr)oxides), show that Cu is not preferentially associated with these mineral phases. Instead, Cu hotspots are associated with particulate organic matter. These observations suggest modification of current sampling protocols, and design of ecotoxicological experiments involving microorganisms, for contaminated soils.     

HXT5 expression is determined by growth rates in Saccharomyces cerevisiae

In the yeast Saccharomyces cerevisiae, hexose transporter (Hxt) proteins transport glucose across the plasma membrane. The Hxt proteins are encoded by a multigene family with 20 members, of which Hxt1-4p and Hxt6-7p are the major hexose transporters. The remaining Hxt proteins have other or unknown functions. In this study, expression of HXT5 under different experimental set-ups is determined. In glucose-grown batch cultures, HXT5 is expressed prior to glucose depletion. Independent of the carbon source used in batch cultures, HXT5 is expressed after 24 h of growth and during growth on ethanol or glycerol, which indicates that growth on glucose is not necessary for expression of HXT5. Increasing the temperature or osmolarity of the growth medium also induces expression of HXT5. In fed-batch cultures, expression of HXT5 is only observed at low glucose consumption rates, independent of the extracellular glucose concentration. The only common parameter in these experiments is that an increase of HXT5 expression is accompanied by a decrease of the growth rate of cells. To determine whether HXT5 expression is determined by the growth rate, cells were grown in a nitrogen-limited continuous culture, which enables modulation of only the growth rate of cells. Indeed, HXT5 is expressed only at low dilution rates. Therefore, our results indicate that expression of HXT5 is regulated by growth rates of cells, rather than by extracellular glucose concentrations, as is the case for the major HXTs. A possible function for Hxt5p and factors responsible for increased expression of HXT5 upon low growth rates is discussed.     

Quality of comminuted sausages formulated from mechanically deboned poultry meat

Comminuted sausages formulated with mechanically deboned poultry meat-MDPM (turkey or chicken, frame or neck) treated in different ways before production (vacuum packed MDPM or air packed skeletons deboned at production) and stored frozen for 6 or 18 weeks have been studied using a full-factorial design and chemical, physical and sensory analysis. MDPM was obtained from a Beehive separator. Comminuted sausages with MDPM from turkey frames, stored at -25?°C as skeletons, packed in air for 18 weeks developed marked rancid odour and flavour which could also be detected by thiobarbituric acid reactive substances (TBARS) and the concentration of volatile compounds. Vacuum packed mechanically deboned meat irrespective of species and carcass part could be stored for up to 18 weeks and used in formulation of sausages without any serious changes in their sensory profiles. Instrumental colour differences between sausages were mainly due to species. Turkey sausages were darker, redder and less yellow than chicken sausages. Moisture and fat contents were influenced by carcass part and to a lesser extent depended on species and storage time. Principal least squares regression (PLS2) showed close relationships between instrumental parameters and sensory attributes. TBARS, hexanal and volatile compounds were highly correlated with rancid flavour while redness (a(?)) was strongly related to sensory colour hue and colour strength.     

Hydrogenation behavior on 30 nm vanadium thin films with two different adhesion conditions

The influence of a thin polycarbonate de-adhesion layer on the hydrogen concentration is studied on 30 nm vanadium films deposited on glass substrates, using electrochemical hydrogenography in an optical microscopy and scanning tunneling microscopy. It is shown that the optical reflection provides information about the de-adhesion morphology (buckles) while the optical transmission signal gives information about concentration and film thickness changes. Artificially patterned samples allow simultaneous studying adhered and de-adhered film parts, for similar mean concentrations. The optical data clearly show a different hydrogen behavior of the two parts. Data interpretation suggests higher local hydrogen content in the adhered film parts than for the detached films parts. Strong changes in the optical transmission of the adhered film parts can be attributed to strong morphological changes at the film surface. These changes are mainly attributed to grain sliding processes in the vanadium film.     

FEM simulation supported evaluation of a hydrogen grain boundary diffusion coefficient in MgH2

The diffusion coefficient data of hydrogen in the Magnesium-hydrogen system shows a large scatter, their trends extrapolations vary at room temperature between 10^?12 m²/s and 10^?29 m²/s. At room temperature the hydrogen diffusion coefficient in MgH₂ is, thus, uncertain by about 17 orders of magnitude. This may be partially attributed to grain boundaries contributing to the measured diffusion coefficient. In this paper we use finite-element (FEM) simulations to evaluate the influence of the grain boundary diffusion on the measured total diffusion depending on the difference of the grain boundary (D_GB) and volume (D_V) diffusion coefficients, as well as on the grain size. These results will be compared to Harrisson's analytical solutions. When the diffusion coefficients differ by more than D_V < 10^?3·D_GB, Harrison's diffusion regime C becomes the best way to describe the total diffusion. The results are used to re-interpret literature data on hydrogen diffusion in MgH₂ from this grain boundary contribution point of view. At 300 K, a hydrogen grain boundary diffusion coefficient ranging from D_GB = 10^?17 m²/s to D_GB = 10^?20 m²/s, depending on the individual type of sample in MgH₂, results from the data evaluation.     

Grouped Gaussian processes for solar power prediction

We consider multi-task regression models where the observations are assumed to be a linear combination of several latent node functions and weight functions, which are both drawn from Gaussian process priors. Driven by the problem of developing scalable methods for forecasting distributed solar and other renewable power generation, we propose coupled priors over groups of (node or weight) processes to exploit spatial dependence between functions. We estimate forecast models for solar power at multiple distributed sites and ground wind speed at multiple proximate weather stations. Our results show that our approach maintains or improves point-prediction accuracy relative to competing solar benchmarks and improves over wind forecast benchmark models on all measures. Our approach consistently dominates the equivalent model without coupled priors, achieving faster gains in forecast accuracy. At the same time our approach provides better quantification of predictive uncertainties.

     

The Next Generation of Colloidal Probes: A Universal Approach for Soft and Ultra‐Small Particles

The colloidal probe technique, which is based on the atomic force microscope, revolutionizes direct force measurements in many fields, such as interface science or biomechanics. It allows for the first time to determine interaction forces on the single particle or cell level. However, for many applications, important “blind spots” remain, namely, the possibility to probe interaction potentials for nanoparticles or complex colloids with a soft outer shell. Definitely, these are colloidal systems that are currently of major industrial importance and interest from theory. The here‐presented novel approach allows for overcome the aforementioned limitations. Its applicability has been demonstrated for 300 nm sized carboxylate‐modified latex particles as well as sub‐micron core–shell particles with a soft poly‐N‐isopropylacrylamide hydrogel shell and a rigid silica core. For the latter, which until now cannot be studied by the colloidal probe technique, determined is the temperature dependency of electrosteric and adhesion forces has been determined on the single particle level.