Quantitative detection of trichloroacetic acid in human urine using isotope dilution high-performance liquid chromatography-electrospray ionization tandem mass spectrometry

The chemical disinfection of drinking water to control microbial contaminants results in the formation of disinfection byproducts (DBPs). The volatile trihalomethanes and the nonvolatile haloacetic acids (HAAs) are the most prevalent DBPs. It is important to monitor human exposure to HAAs because of their potential adversehealth effects, such as cancer. Among the HAAs, urinary trichloroacetic acid (TCAA) is a potential valid biomarker for assessing chronic ingestion exposure to HAAs from drinking water. We have developed a rugged, high-throughput, sensitive, accurate, and precise assay for the measurement of trace levels of TCAA in human urine using a simple solid-phase extraction (SPE) cleanup followed by isotope dilution high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). TCAA is extracted from the urine using SPE, separated from other extract components by reversed-phase HPLC, and analyzed by negative ion electrospray ionization-isotope dilution-MS/MS using a multiple reaction monitoring experiment. The method is simple and fast and is not labor intensive (sample preparation and analysis can be performed in approximately 15 min) with a limit of detection of 0.5 ng/mL in 1 mL of urine.     

Composition and quality attributes of conventionally and organically farmed Pangasius fillets (Pangasius hypophthalmus) on the German market

A range of conventionally and organically farmed Pangasius or sutchi catfish fillets available on the German market were analysed to compare both composition and quality. Differentiation of Pangasius hypophthalmus from Pangasius bocourti was achieved by RFLP-SSCP analysis. The protein content of conventionally farmed fillets ranged between 13.3 and 15.7%, whereas organically produced fillets had significantly higher protein contents of between 17.0 and 17.4%. No difference in the fat content between farming methods was observed, which varied between 1.4 and 3.2%. Polyunsaturated fatty acids represented about 24% of the total fatty acids with a high level of linoleic acid. The comparison of the proximate composition indicated that water was added to most of the conventionally farmed products, in different amounts, as well as water-binding capacity enhancing additives. Differential scanning calorimetry was used to demonstrate the presence of polyphosphate on muscle proteins. Differences in texture, water-binding capacity and colour are discussed.     

Photocaged Hoechst Enables Subnuclear Visualization and Cell Selective Staining of DNA in vivo

Selective targeting of DNA by means of fluorescent labeling has become a mainstay in the life sciences. While genetic engineering serves as a powerful technique and allows the visualization of nucleic acid by using DNA-targeting fluorescent fusion proteins in a cell-type- and subcellular-specific manner, it relies on the introduction of foreign genes. On the other hand, DNA-binding small fluorescent molecules can be used without genetic engineering, but they are not spatially restricted. Herein, we report a photocaged version of the DNA dye Hoechst33342 (pcHoechst), which can be uncaged by using UV to blue light for the selective staining of chromosomal DNA in subnuclear regions of live cells. Expanding its application to a vertebrate model organism, we demonstrate uncaging in epithelial cells and short-term cell tracking in vivo in zebrafish. We envision pcHoechst as a valuable tool for targeting and interrogating DNA with precise spatiotemporal resolution in living cells and wild-type organisms.     

Synthesis and isolation of <Emphasis Type="Italic">trans</Emphasis>-7, <Emphasis Type="Italic">cis</Emphasis>-9 octadecadienoic acid and other CLA isomers by base conjugation of partially hydrogenated γ-linolenic acid

CLA is of considerable interest because of reported potentially beneficial effects in animal studies. CLA, while not yet unambiguously defined, is a mixture of octadecadienoic acids with conjugated double bonds. The major isomer in natural products is generally considered to be cis-9,trans-11-octadecadienoic acid (c9, t11), which represents >75% of the total CLA in most cases. Other isomers are drawing increased attention. The t7,c9 isomer, which is often the second-most prevalent CLA in natural products, has been reported to represent as much as 40% of total CLA in milk from cows fed a high-fat diet. The need for a reference material became apparent in a recent study directed specifically at measuring t7,c9-CLA in milk, plasma, and rumen. A suitable standard mixture was produced by stirring 0.5 g of γ-linolenic acid (all cis-6,9, 12-C_18∶3) with 100 mL of 10% hydrazine hydrate in methanol for 2.5 h at 45°C. The solution was diluted with H₂O and acidified with HCl. The resulting partially hydrogenated FA were extracted with ether/petroleum ether, dried with Na₂SO₄, and conjugated by adding of 6.6% KOH in ethlylene glycol and heating for 1.5 h at 150–160°C. Approximately 20 mg each of cis-6, trans-8; trans-7, cis-9; cis-9, trans-11; and rans-10, cis-12 were obtained along with other FA. Methyl esters (FAME) of these four cis/trans isomers were resolved by Ag⁺HPLC (UV 233) and partially resolved by GC/(MS or FID) (CP-Sil 88). Treatment of these FAME with I₂ yielded all possible cis/trans (geometric) isomers for the four positions 6,8; 7,9; 9,11; and 10,12.     

Stress Analysis of Elastic-Plastic Contact Damage in Ceramic Coatings on Metal Substrates

An elastic-plastic analysis of damage in ceramic coatings on metal substrates from contacts with spherical indenters is made using finite element modeling. Computations are carried out specifically for plasma-sprayed alumina:titania on a soft steel. The algorithm assumes an elastic sphere with frictionless contact on a flat elastic-plastic layered specimen, and incrementally evaluates the expanding contact field as a function of applied load. Two key aspects of the contact field are examined: (i) the indentation stress-strain curve; (ii) the damage zone geometry. Composite coating/substrate indentation stress-strain curves are computed for two coating thicknesses, using input material parameters from iterative fits to data from control tests on free-standing coating and substrate materials. Contours of principal shear stresses, most notably those contours corresponding to yield zone boundaries in both the softer substrate and the harder coating, are mapped out in the fully plastic region. Corresponding distributions of tensile stresses are also mapped out, and are shown to correlate with the locations of transverse fractures in the coating. General implications concerning material and geometrical design of ceramic-based layer structures are discussed.     

Microbial Fuel Cell Application for Azoic Dye Decolorization with Simultaneous Bioenergy Production Using Stenotrophomonas sp.

A single‐chamber air‐cathode microbial fuel cell (MFC) was successfully applied for decolorization of the diazoic dye Reactive Black 5 (RB5) with simultaneous production of electricity. An innovative low‐cost medium, the marine water‐glucose‐yeast extract medium (MWGY), was developed which appears as a textile marine effluent in terms of salinity. The anode compartment containing a single bacterial strain of Stenotrophomonas sp. showed almost complete dye decolorization with different RB5 concentrations after seven days of treatment. For the four dye concentrations used, a correlation between dye removal and production of electrical energy was found. The comparison of the decolorization process in the MFC with that in a batch reactor as control experiment highlights the efficiency of the single‐chamber air‐cathode MFC technology which improved the dye removal by the same bacterial strain.     

Retinal neurostimulator for a multifocal vision prosthesis.

A neurostimulator application-specific integrated circuit (ASIC) with scalable circuitry that can stimulate 14 channels, has been developed for an epi-retinal vision prosthesis. This ASIC was designed to allow seven identical units to be connected to control up to 98 channels, with the ability to stimulate 14 electrodes simultaneously. The neurostimulator forms part of a vision prosthesis, designed to restore vision to patients who have lost their sight due to retinal diseases such as retinitis pigmentosa and macular degeneration. For charge balance, the neurostimulator was designed to stimulate with current sources and sinks operating together, and with the ability to drive a hexagonal mosaic of electrodes to reduce the electrical crosstalk that occurs when multiple bipolar stimulation sites are active simultaneously. A hexagonal mosaic of electrodes surrounds each stimulation site and has been shown to effectively isolate each site, increasing the ability to inject localized independent charge into multiple regions simultaneously.     

Interactive visualization for opportunistic exploration of large document collections

Finding relevant information in a large and comprehensive collection of cross-referenced documents like Wikipedia usually requires a quite accurate idea where to look for the pieces of data being sought. A user might not yet have enough domain-specific knowledge to form a precise search query to get the desired result on the first try. Another problem arises from the usually highly cross-referenced structure of such document collections. When researching a subject, users usually follow some references to get additional information not covered by a single document. With each document, more opportunities to navigate are added and the structure and relations of the visited documents gets harder to understand.     

Fabrication of bronze components by metal injection moulding using powders with different particle characteristics

The aim of this work is to fabricate bronze components by metal injection moulding (MIM) studying the possibility of changing partially or totally the gas atomised powder by water atomised ones that are cheaper than the former. In order to carry out this study, a bronze 90/10 gas atomised spherical powder (usual MIM powder <22 μm) and two water atomised irregular powders (particle size <35 μm and <140 μm) were mixed in different proportions. As received powders and their mixtures were used to fabricate feedstocks and processed by MIM to evaluate the influence of powder particle size and morphology on debinding and sintering stages. Finally, both mechanical properties hardness and maximum flexural stress were determined to characterize the sintered materials. The addition of irregular fine and coarse powders was found to affect the moulding process, although densities and mechanical properties close to values of gas atomised one were obtained after sintering. Therefore, the use of water atomised bronze powders could be a promising way to diminish production costs in this technology.