Cellular differentiation of non-transformed intestinal epithelial cells is regulated by Lactobacillus rhamnosus and L. casei strains.

The aim of this study was to characterize an in vitro modulating effect of three commensal Lactobacillus strains on cellular differentiation of non-transformed crypt-like rat small intestinal cell line IEC-18. IEC-18 was grown on extracellular matrix, with or without presence of Lactobacillus strains. Gene expression of IEC-18 bacterial detection system - such as Toll-like receptors TLR-2, TLR-4, signal adapter MyD88, cytoplasmic NOD2 receptor, inflammatory cytokines IL-18, IL-1beta, chemokine IL-8 and enzyme caspase-1 - was evaluated using real-time PCR. Expression and localization of TLR-2, TLR-4, IL-18 and caspase-1 proteins was demonstrated by Western blotting and immunofluorescent staining. Secretion of IL-18 to apical and basolateral surfaces was assayed by ELISA. Our results suggested that L. casei LOCK0919 accelerated differentiation of IEC-18 by stimulating TLR-2, TLR-4, MyD88, IL-18, caspase-1 mRNAs and proteins. L. casei LOCK0919 increased expression and transfer of villin and beta-catenin from cytoplasm to cell membrane. Presence of L. rhamnosus LOCK0900 resulted in detachment of IEC-18 layer from extracellular matrix leading to induction of IL-1beta, of TLR-2 and IL-8 mRNAs and stimulation of MyD88, caspase-1 and cytosolic receptor NOD2 mRNAs. L. rhamnosus LOCK0908 was not recognized by TLR-2 or TLR-4 receptors. Lactobacilli-IEC-18 crosstalk enhanced immune and barrier mucosal functions.

A new approach to the isolation and characterization of wheat flour allergens.

BACKGROUND: The incidence of food allergy to wheat is increasing. Its diagnosis depends on the purity of major allergens and their inclusion in tests. Isolation and characterization of wheat allergens are therefore of utmost importance. OBJECTIVE: To purify and identify wheat flour allergens most frequently recognized by patients' IgE antibodies and to study their allergenicity. METHODS: Water/salt-soluble extracts from wheat flour were prepared and separated using a combination of ultrafiltration, isoelectric focusing and liquid chromatography. Purified proteins were analysed by immunoblotting using pooled sera from patients with atopic dermatitis who possessed IgE specific to wheat. Wheat proteins found to bind IgE were subsequently identified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. The frequency and intensity of IgE binding of isolated proteins were tested using individual sera from patients and controls. RESULTS: We developed a procedure that allows isolation of wheat allergens from natural sources. Twenty-seven potential wheat allergens have been successfully identified; of these, the following seven are newly reported in food allergy: endogenous α-amylase/subtilisin inhibitor, trypsin/α-amylase inhibitor (AAI) CMX1/CMX3, thaumatin-like protein (TLP), xylanase inhibitor protein-1, ß-glucosidase, class II chitinase and 26 kDa endochitinase. TLP and wheatwin were shown to activate patients' basophils to a similar extent as two well-known allergens, lipid transfer protein (Tri a 14) and AAI 0.19 (Tri a 28.0101). CONCLUSION AND CLINICAL RELEVANCE: Our new approach enables the isolation of water/salt-soluble wheat allergens in their native form in amounts sufficient both for biological testing (in vivo and in vitro) and for physicochemical characterization. Such studies will lead to a more detailed knowledge of allergenicity of wheat proteins and to improved accuracy of diagnostic tests.

Effects of signalisation on four-arm urban junction safety.

This paper investigates the effect of signalisation on safety of urban priority controlled four-arm junctions. Forty-eight such junctions were carefully selected to have different values for the geometric and traffic characteristics that may influence the safety effects of signalisation. Long periods before and after signalisation were considered so that the effect of the regression to the mean phenomenon becomes negligible. Signalisation was found either to increase the junction safety or to leave it unchanged. Discriminant analysis showed that among numerous variables considered those significantly influencing the effects of signalisation on safety are the flow weighted average width per arm, the flow weighted average grade per stream, and a function of the product of the conflicting stream flows of the junction. The discriminant function developed predicts quite satisfactorily the behaviour of junction safety after signalisation.

Establishing relationships between accidents and flows at urban priority road junctions.

This papers explores the effects of traffic stream flows on accident potential at urban priority-controlled (i.e. unsignalized), four-arm junctions. Forty-three urban priority junctions were carefully selected so that other than flow parameters expected to influence accident potential have similar values at the junctions considered. Using traffic accident data for five-year time periods and the corresponding 24-hour flows, a new exposure index is proposed consisting of an expression of the flows of the junction's interacting traffic streams. The regression of this exposure index on the expected number of accidents per year at junctions of the type examined yields a quite satisfactory correlation coefficient, better than those achieved when other proposed indices are used.

[Simultaneous determination of volatile substances in intact apples and apple juice using gas chromatographic head-space analysis]. / Simultane Bestimmung flüchtiger, von unversehrten Apfeln abgegebener und im Apfelsaft befindlicher Stoffe mit Hilfe der gaschromatographischen Head-space-Analyse.

Relations between the rate of the generation of volatiles by intact apples during their ripening and the contents of the volatiles in the juice pressed out of the apples were investigated with the aid of two different techniques of headspace gas analysis. The juice was analyzed by the technique of stripping and trapping in a closed circuit, which made it possible to determine the violatiles in concentrations of several tens of micrograms/1 of juice. The volatiles generated by intact apples were determined by analysis the gas percolating through a layer of fruits by a sample-enrichment technique, the sensitivity of analysis being some hundredths of micrograms/1. The resistance of the apple matrix against the transfer of volatiles generated by apples to their surrounding does not vary substantially during the ripening of apples on tree. The permeability constant of the apple matrix ranges from some tenths to units of m1/(kg . h) with all the volatiles and apple species studied.