Specific,semi-quantitative detection of the soybean allergen Gly m Bd 30K DNA by PCR

A semi-quantitative PCR-based system has been developed for detection of DNA sequences for the soybean allergen Gly m Bd 30K. The selected primers were highly specific for soybean and did not show amplification from a panel of legume relatives. Repeatability was assessed in a spiking experiment of soybean in wheat flour (0.0001–100% soybean), using known standards for comparison of the amount of output DNA from different PCR reactions. The frequency of PCR reactions with successful amplification of the soybean allergen sequence was highly dependent on initial target DNA concentration, and showed a rapid sigmoidal decrease, when target concentration approached the detection limit (0.01%). For samples with successful amplification there was a good correlation between the initial amount of soybean in the mixture and the output from PCR, when suitable block designs were used to control experimental errors. The simple approach used for quantification in this study proved efficient for assessment of homogeneity of self-prepared soybean bars used for provocation tests of food allergic patients in clinical practice. In addition the method was efficient in detecting soybean allergen sequences in a number of processed foods.     

Self-assembled silver nanoprisms monolayers at the liquid/liquid interface

A crown ether derivative (4′-aminobenzo-15-crown-5 hydrotetrafluoroborate) can mediate the self-assembly of silver nanoprisms at the liquid/liquid interface in the form of a stable hybrid nanocomposite film. The metallic luster of the interfacial film results from the electronic coupling of Ag nanoprisms, suggesting the formation of closely packed nanoprism thin films. The interfacial film of nanoprisms could be transferred to solid substrates as a Langmuir–Blodgett film. The properties of films were studied by UV–vis spectroscopy and transmission electron microscopy.     

A reliable and sensitive time-resolved fluorescent immunochromatographic assay (TRFICA) for ochratoxin A in agro-products

Immunochromatographic assays (ICAs) are considered as a suitable diagnostic tool for the detection of mycotoxins. Mycotoxins and especially, ochratoxin A are analytes with more demanding sensitivity requirements. To enhance the sensitivity of current immunochromatographic assays for ochratoxin A (OTA), a novel sensitive ICA was developed in this study. In the assay, microspheres enclosing fluorescent europium (III) [Eu(III)] nanoparticles (EuNPs) were used as a label for OTA monoclonal antibody (OTA-mAb) conjugation. Accordingly, assay was called time-resolved fluorescent immunochromatographic assay (TRFICA). The test strip was composed of three parts: a sample pad, nitrocellulose membrane and an absorbent pad. As for detection, a proper concentration of conjugated microspheres was pipetted into the microtube and sample extract was added to it. Then the strip was inserted into the tube and the fluid flow along the strip. The TRFICA results were obtained in 8 min and read by a portable TRFICA strip reader. The established method allows quantitative determination of OTA with limit of detection as low as 1.0 μg kg⁻¹ in the samples. For validation, spiked samples including wheat, maize, soybean and rice were respectively assayed by TRFICA and a standard high performance liquid chromatography equipped with a fluorescence detector (HPLC-FLD), and good agreement of results was obtained between two methods.     

Rapid detection method for aflatoxin B1 in soybean sauce based on fluorescent microspheres probe

Soybean sauce, a Chinese traditional and daily condiment, is often contaminated by aflatoxin B₁. An extract-free immunochromatographic assay was proposed based on fluorescent microspheres probe for the' detection of aflatoxin B₁ in soybean sauce. The probe was prepared by coupling fluorescent microspheres with anti-aflatoxin B₁ antibody by the 1-ethyl-3(3-dimethylaminopropyl) carbodiimides hydrochloride-mediated method. The background from the soybean sauce sample on strip was eliminated because of the optical property of the probe. The sample without extracting procedure was directly detected by diluting with 10% methanol solution. The visible detection limit for the qualitative analysis of aflatoxin B₁ in the proposed method was 2.5 μg/L, which was lower than the maximum level of 5 μg/L set by the Chinese government. The results were well agreed with those obtained by liquid chromatography-mass spectrometry (LC-MS). The method showed satisfactory characteristics, such as rapid detection, easy operation, and high sensitivity, and can thus be applied for the large-scale and on-site screening of soybean sauce contaminated with aflatoxin B₁. To our knowledge, this report is the first one on the qualitative detection of aflatoxin B₁ in dark colored food samples directly by fluorescent microspheres probe-based immunochromatography.     

The ALEPH minivertex detector

Vertex detectors allow high precision reconstruction of particle tracks and therefore make possible the investigation of the decay topology of short-lived particles in collider experiments.In the ALEPH experiment at LEP a minivertex detector will be installed. It consists of silicon microstrip detectors arranged on two concentric “cylindrical” surfaces around the interaction point. With this geometry it will be possible to measure the r − ϕ − z coordinates of particles traversing the detector. The expected position resolution is 10 μm in r − ϕ and 20 μm in r − z.For optimum signal processing monolithic CMOS readout electronics are under development. Each chip consists of 60 charge sensitive preamplifiers, multiplexed into one output channel. Fast power switching will reduce heat dissipation.Details about construction and expected device performance will be described.     

Counting anticoincidences to reduce statistical uncertainty in the calibration of a multiplicity detector

The observation of anticoincidences reduced statistical uncertainties as well as pileup corrections in the efficiency calibration of a multiplicity detector. The efficiency of the 4π gadolinium-loaded organic scintillation detector was measured for detecting capture of neutrons scattered by the hydrogen in a second detector. In such situations the effect of background upon the counting fluctuations is reduced below the value that would occur if the average counting rate during coincidence gates were utilized. The applicable formulae were obtained by elementary methods and were verified with experimental data and with computer simulations for a broad range of detector efficiencies and background intensities. The general requirements for use of the technique are indicated.     

Conformations of aniline black (polyaniline) molecules

Conformations of aniline black molecules were studied using a semi-empirical computer-aided analysis. A strong deformation of the ‘planar structure’ of the molecules has been found.     

The Tumor Suppressor Roles of miR-433 and miR-127 in Gastric Cancer

The discovery of microRNAs (miRNAs) provides a new and powerful tool for studying the mechanism, diagnosis and treatment of human cancers. Currently, the methylation epigenetic silencing of miRNAs with tumor suppressor features by CpG island hypermethylation is emerging as a common hallmark of different tumors. Here we showed that miR-433 and miR-127 were significantly down-regulated in gastric cancer (GC) tissues compared with the adjacent normal regions in 86 paired samples. Moreover, the lower level of miR-433 and miR-127 was associated with pM or pTNM stage in clinical gastric cancer patients. The restored expression of miR-433 and miR-127 in GC cells upon 5-Aza-CdR and TSA treatment suggested the loss of miR-433 and miR-127 was at least partly regulated by epigenetic modification in GC. Furthermore, the ectopic expression of miR-433 and miR-127 in gastric cancer cell lines HGC-27 inhibits cell proliferation, cell cycle progression, cell migration and invasion by directly interacting with the mRNA encoding oncogenic factors KRAS and MAPK4 respectively. Taken together, our results showed that miR-433 and miR-127 might act as tumor suppressors in GC, and it may provide novel diagnostic and therapeutic options for human GC clinical operation in the near future.