Application of polymerase chain reaction–restriction fragment length polymorphism analysis and lab‐on‐a‐chip capillary electrophoresis for the specific identification of game and domestic meats

BACKGROUND: The objective of this study was to adapt and improve previously published polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) methods aimed at the identification of game and domestic meats, by replacing the gel electrophoretic steps for DNA fragment analysis by a chip‐based capillary electrophoresis system. RESULTS: PCR amplification of a mitochondrial 12S rRNA gene fragment and subsequent digestion of the amplicons with either MseI or a combination of MboII, BslI, and ApoI endonucleases generated characteristic PCR‐RFLP profiles that allowed discrimination among ten relevant game and domestic meat species. The Agilent 2100 Bioanalyzer utilises capillary electrophoresis on a microchip device that is capable of rapidly sizing DNA fragments, offering a valuable recent development for the analysis of complex DNA banding patterns. CONCLUSION: Results obtained in this work indicated that banding resolution on the system was sensitive, with detection of some small DNA fragments that were not observed with the published conventional PCR‐RFLP gel‐based method. Therefore, the new faster and easy handling procedure provides an additional powerful tool that can be employed for meat speciation. Copyright © 2009 Society of Chemical Industry     

Electron paramagnetic resonance spectroscopy studies of oxidative degradation of an active pharmaceutical ingredient and quantitative analysis of the organic radical intermediates using partial least-squares regression

Electron paramagnetic resonance (EPR) spectroscopy was used to study the radical species formed during the oxidation of an active pharmaceutical ingredient in the solid state. It was found that the extent of radical generation correlated to the formation of an oxidative degradation product. Multifrequency EPR and electron nuclear double resonance spectroscopy gave additional information on the identity of the organic radical species involved in the oxidation process, and a mechanism was proposed for the degradation, involving the formation of both carbon-centered and peroxy radicals. The multivariate analysis technique of partial least-squares (PLS) regression was then used to determine the extent of oxidation of the active pharmaceutical ingredient from the EPR spectra. The suitability of this approach was demonstrated from its application to a series of standards. The conventional approach for the quantitative analysis of EPR spectra is to measure the peak height or to perform double integration of the spectral region containing the signal of interest. Both of these methods have intrinsic errors associated with them, particularly for weak EPR signals with a poor signal-to-noise ratio or a sloping background response. The results obtained showed that greatly improved quantitation was obtained using the PLS regression approach.     

Fabrication and characterization of epitaxial Ba0.7Sr0.3TiO3 thin films for optical waveguide applications

The optical properties of barium strontium titanate (Ba0.7Sr0.3TiO3; BST) thin films are described. The BST thin films were epitaxially grown upon MgO (001) substrates by pulsed laser deposition. The crystallographic properties of the BST thin films were examined by x-ray diffraction. The BST thin films were highly optically transparent in the visible region. The optical waveguide properties were characterized by a prism coupling technique. An inverse-WBK method was employed to determine the refractive-index profile along the thickness of the BST films. Optical losses were measured by a moving fiber method, and the optical losses were found to be 0.93 dB/cm for the TE0 mode and 1.29 dB/cm for the TM0 mode at 1550 nm. Electro-optic (E-O) properties were measured by a phase-modulation detection method at 632.8 nm, and the BST films exhibited a predominantly quadratic E-O effect with a quadratic E-O coefficient of 6.64 x 10(-18) m2/V2.     

An efficient automated z-shim based method to correct through-slice signal loss in EPI at 3T

Object  To develop an efficient, automated method to correct through-slice signal loss in gradient-echo EPI at 3T. Material and methods  The optimal choice of two z-shim values for signal recovery was determined from simulations and experiments. The specific required z-shim values are determined using a rapid calibration method that combines information about the slice profile with a sparse set of measurements. The proposed correction method was implemented for a language fMRI study which suffers from signal loss near the auditory canals, and tested on 12 volunteers. Results  Using a square root sum of squares combination of two z-shim values full signal restoration (to within 2% of the correct value) was achieved in 96% of all correctable brain pixels for 3 mm slices, and partial correction in pixels outside this range. In all subjects, language processing activation was recovered in the inferior and lateral areas of the left temporal lobe which was not detectable with conventional fMRI. Conclusion  The careful choice of two z-shim values by the proposed method achieves through-slice signal loss correction for the majority of pixels in the brain for 3 mm slices at 3T.     

MiR-126-3p Is Dynamically Regulated in Endothelial-to-Mesenchymal Transition during Fibrosis

In fibrotic diseases, myofibroblasts derive from a range of cell types including endothelial-to-mesenchymal transition (EndMT). Increasing evidence suggests that miRNAs are key regulators in biological processes but their profile is relatively understudied in EndMT. In human umbilical vein endothelial cells (HUVEC), EndMT was induced by treatment with TGFβ2 and IL1β. A significant decrease in endothelial markers such as VE-cadherin, CD31 and an increase in mesenchymal markers such as fibronectin were observed. In parallel, miRNA profiling showed that miR-126-3p was down-regulated in HUVECs undergoing EndMT and over-expression of miR-126-3p prevented EndMT, maintaining CD31 and repressing fibronectin expression. EndMT was investigated using lineage tracing with transgenic Cdh5-Cre-ERT2; Rosa26R-stop-YFP mice in two established models of fibrosis: cardiac ischaemic injury and kidney ureteric occlusion. In both cardiac and kidney fibrosis, lineage tracing showed a significant subpopulation of endothelial-derived cells expressed mesenchymal markers, indicating they had undergone EndMT. In addition, miR-126-3p was restricted to endothelial cells and down-regulated in murine fibrotic kidney and heart tissue. These findings were confirmed in patient kidney biopsies. MiR-126-3p expression is restricted to endothelial cells and is down-regulated during EndMT. Over-expression of miR-126-3p reduces EndMT, therefore, it could be considered for miRNA-based therapeutics in fibrotic organs.     

Relationship between the physicochemical properties of starches and the glycemic indices of some Jamaican yams (Dioscorea spp.)

Starch granules from round leaf yellow yam (RY), Lucea yam (LY), white yam (WY), and Chinese yam (CY) grown in Jamaica were isolated and the relationship between starch amylose content, crystallinity, microscopic properties, in vitro digestibility, and the glycemic index (GI) of the tubers was investigated. The results indicate that RY had the highest amylose content (265.30 +/- 0.09 g/kg starch) while CY the lowest (111.44 +/- 0.03 g/kg starch). A corresponding variation in starch digestibility and GI was also observed, as CY which had the highest in vitro digestibility had the highest GI (21.27 +/- 0.01 and 97.42 +/- 0.62%, respectively), while RY, LY, and WY starches with low digestibility had lowest GI. Differences in the crystalline pattern of the different starches were observed, where RY, LY, and WY displayed the type B crystalline pattern while CY had the intermediate crystallite (type C).     

Radiocarbon-based assessment of fossil fuel-derived contaminant associations in sediments

Hydrophobic organic contaminants (HOCs) are associated with natural organic matter (OM) in the environment via mechanisms such as sorption or chemical binding. The latter interactions are difficult to quantitatively constrain, as HOCs can reside in different OM pools outside of conventional analytical windows. Here, we exploited natural abundance variations in radiocarbon (14C) to trace various fossil fuel-derived HOCs (14C-free) within chemically defined fractions of contemporary OM (modern 14C content) in 13 samples including marine and freshwater sediments and one dust and one soil sample. Samples were sequentially treated by solvent extraction followed by saponification. Radiocarbon analysis of the bulk sample and resulting residues was then performed. Fossil fuel-derived HOCs released by these treatments were quantified from an isotope mass balance approach as well as by gas chromatography-mass spectrometry. For the majority of samples (n = 13), 98-100% of the total HOC pool was solvent extractable. Nonextracted HOCs are only significant (29% of total HOC pool)in one sample containing p,p-2,2-bis(chlorophenyl)-1,1,1-trichloroethane and its metabolites. The infrequency of significant incorporation of HOCs into nonextracted OM residues suggests that most HOCs are mobile and bioavailable in the environment and, as such, have a greater potential to exert adverse effects.