The major soyabean allergen P34 resists proteolysis in vitro and is transported through intestinal epithelial cells by a caveolae-mediated mechanism

Soya is considered to be one of the eight most significant food allergens. Among the allergenic soya proteins determined to date, P34 has been identified as one of the immunodominant soya antigens. Sensitisation to a specific food antigen like P34 generally follows the transit of intact antigens across the intestinal barrier and usually occurs in infants, who are most susceptible to food allergies. In the present study, we used the intestinal epithelial cell line IPEC-J2, which was originally derived from the jejunum of a neonatal piglet, to recapitulate the infant intestinal epithelium and study the binding and uptake of P34 protein. P34 was partially resistant to degradation in an in vitro proteolysis assay. IPEC-J2 cells were able to endocytose intact P34, as shown by immunofluorescence and immunoelectronmicroscopy methods. P34 associated with lipid raft microdomains of IPEC-J2 cells, and disruption of caveolae/lipid raft microdomains using methyl-β-cyclodextrin abolished P34 endocytosis, indicating that the observed endocytosis was mediated by caveolae. Using IPEC-J2 cells grown on Transwell filters, we further demonstrated that P34 is transported through the epithelial monolayer by transcytosis. Piglets frequently show hypersensitivity to soya antigens, and in this study, we show that healthy adult pigs with dietary exposure to soya protein mount an antibody response to soyabean protein P34, suggesting that this protein has entered the body, probably through gastrointestinal uptake. In summary, our data suggest that soya P34 resists proteolysis in the gastrointestinal tract and is transported through the intestinal epithelial barrier, thereby allowing sensitisation of immune cells in the sub-epithelial compartment. © Cambridge University Press 2012 [accessed November 30th 2012

Influence of microwave probes on calibrated on-wafer measurements

On-wafer probing with ground-signal-ground (GSG) probes contributes a variety of side effects, which are related to the measured line type, the carrier material, the layout with the neighboring structures, and the probe. Thus, the size and shape of the probe together with the measured line type and the neighboring circuits influence the quality of the calibrated measured result. This paper presents corresponding results when using the multiline-thru-reflect-line (mTRL) calibration, which is commonly accepted as one of the most accurate calibration algorithms, and concentrates on the impact of the probe construction together with neighboring elements, for the most common planar transmission lines, coplanar waveguides (CPWs), and thin-film microstrip lines (TFMSLs). For the first time, design guidelines with regard to the layout, the measurement environment, and the construction of the probes are given

Disposable micro-fluidic biosensor array for online parallelized cell adhesion kinetics analysis on quartz crystal resonators

In this contribution we present a new disposable micro-fluidic biosensor array for the online analysis of adherent Madin Darby canine kidney (MDCK-II) cells on quartz crystal resonators (QCRs). The device was conceived for the parallel cultivation of cells providing the same experimental conditions among all the sensors of the array. As well, dedicated sensor interface electronics were developed and optimized for fast spectra acquisition of all 16 QCRs with a miniaturized impedance analyzer. This allowed performing cell cultivation experiments for the observation of fast cellular reaction kinetics with focus on the comparison of the resulting sensor signals influenced by different cell distributions on the sensor surface. To prove the assumption of equal flow circulation within the symmetric micro-channel network and support the hypothesis of identical cultivation conditions for the cells living above the sensors, the influence of fabrication tolerances on the flow regime has been simulated. As well, the shear stress on the adherent cell layer due to the flowing media was characterized. Injection molding technology was chosen for the cheap mass production of disposable devices. Furthermore, the injection molding process was simulated in order to optimize the mold geometry and minimize the shrinkage and the warpage of the parts. MDCK-II cells were cultivated in the biosensor array. Parallel cultivation of cells on the gold surface of the QCRs led to first observations of the impact of the cell distribution on the sensor signals during cell cultivation. Indeed, the initial cell distribution revealed a significant influence on the changes in the measured acoustic load on the QCRs suggesting dissimilar cell migrations as well as proliferation kinetics of a non-confluent MDCK-II cell layer