Epithelial transport and deamidation of gliadin peptides: a role for coeliac disease patient immunoglobulin A.

In coeliac disease, the intake of dietary gluten induces small-bowel mucosal damage and the production of immunoglobulin (Ig)A class autoantibodies against transglutaminase 2 (TG2). We examined the effect of coeliac patient IgA on the apical-to-basal passage of gluten-derived gliadin peptides p31-43 and p57-68 in intestinal epithelial cells. We demonstrate that coeliac IgA enhances the passage of gliadin peptides, which could be abolished by inhibition of TG2 enzymatic activity. Moreover, we also found that both the apical and the basal cell culture media containing the immunogenic gliadin peptides were able to induce the proliferation of deamidation-dependent coeliac patient-derived T cells even in the absence of exogenous TG2. Our results suggest that coeliac patient IgA could play a role in the transepithelial passage of gliadin peptides, a process during which they might be deamidated.

Randomised clinical trial: low-FODMAP rye bread vs. regular rye bread to relieve the symptoms of irritable bowel syndrome.

BACKGROUND: Grains are high in FODMAPs (Fermentable Oligo-, Di-, Monosaccharides And Polyols) and often considered as triggers of IBS symptoms. AIM: To evaluate if rye bread low in FODMAPs would be better tolerated than regular rye bread in subjects with IBS. METHODS: The study was conducted as a randomised double blind controlled cross-over study (n = 87). Participants were supplied with both regular rye bread and low-FODMAP rye bread for 4 weeks. Symptoms were measured with a symptom severity scoring system (IBS-SSS) and visual analogue scale (VAS) assessments of individual symptoms. Quality of life was monitored. Colonic fermentation was measured by the breath hydrogen test and dietary intake by food diaries. RESULTS: Dietary fibre intake increased during both study periods compared to baseline. Many signs of IBS i.e. flatulence, abdominal pain, cramps and stomach rumbling were milder on the low-FODMAP rye bread (P-values: 0.04; 0.049; 0.01 and 0.001). The mean of VAS measurements was favourable towards LF bread [-3 (95% CI): -6 to -1, P = 0.02] but no differences were detected in IBS-SSS or quality of life. The AUC of breath hydrogen values was significantly lower during the low-FODMAP bread period (median 52.9 vs. 72.6; P = 0.01). CONCLUSIONS: Low-FODMAP rye bread helps IBS patients to control their symptoms and reduces gastrointestinal gas accumulation. However, replacing regular rye bread by low-FODMAP bread without concomitant broader dietary changes does not improve quality of life or IBS-SSS. Nonetheless, inclusion of low-FODMAP rye bread in diet might be one way that IBS patients could increase their fibre intake.

Characterisation of hydrolysable tannins from leaves of Betula pubescens by high-performance liquid chromatography-mass spectrometry.

A high-performance liquid chromatography-electrospray ionisation mass spectrometry (HPLC-ESI-MS) method, assisted by diode array detection, for the characterisation of individual hydrolysable tannins in birch leaves was developed. With the method, it was found that birch (Betula pubescens) leaves contained an exceptionally complex mixture of hydrolysable tannins; 14 gallotannins and 20 ellagitannins were identified. The developed HPLC-ESI-MS method allows the qualitative and quantitative determination of individual gallotannins and ellagitannins directly from crude birch leaf extract. This is important in studying ecological functions of these phenolic compounds, especially their role in the resistance of birch leaves against insects.

Phenolics and betacyanins in red beetroot (Beta vulgaris) root: distribution and effect of cold storage on the content of total phenolics and three individual compounds.

The distribution of total phenolics and main betacyanins in red beetroot (Beta vulgaris) root was determined. Also, the subsequent effects of cold storage on the content of total phenolics, main betacyanins (betanin and isobetanin), and the main known ferulic acid ester (beta-D-fructofuranosyl-alpha-D-(6-O-(E)-feruloylglucopyranoside) were determined in the peel, which is the root part containing the largest amount of total phenolics. The content of total phenolics in the red beetroot water extracts was determined according to a modification of the Folin-Ciocalteu method and expressed as gallic acid equivalents (GAE). The compounds of interest were identified by HPLC-ESI-MS and NMR techniques, and the contents of compounds were determined by HLPC analyses. The total phenolic contents in various root parts were found to decrease in the order peel, crown, flesh. Significant differences in the contents of total phenolics and individual compounds were found when the effect of cold storage (5 degrees C, 0-196 days) on the constituents of the peel from intact roots was examined. In addition to the betacyanins of red beetroot peel found in our earlier study, tentative identifications of betanidin and feruloylamaranthin were made.

Patterns in content of phenolic compounds in leaves of mountain birches along a strong pollution gradient.

The contents of individual low-molecular weight phenolic compounds (LMWPs) in mountain birch, Betula pubescens ssp. czerepanovii, leaves collected during 1996-1998 in six plots 7-65 km south of the nickel-copper smelter at Monchegorsk, Kola Peninsula, NW Russia, were reported. A high-performance liquid chromatography-electrospray ionisation-mass spectrometry (HPLC-ESI-MS) was used for the rapid identification of low-molecular weight phenolics. Quantification was performed by the analytical high-performance liquid chromatography with UV-detection. Contents of (+)-catechin and some gallic acid derivatives decreased significantly, and contents of flavonol glycosides slightly increased with the distance from the smelter. Hydroxycinnamic acid derivatives remained unaffected. These changes in birch leaf phenolics are probably related to the effect of environmental contamination on the biosynthetic reactions both in the shikimate and phenylpropanoid pathways.

Betalains and phenolics in red beetroot (Beta vulgaris) peel extracts: extraction and characterisation.

The extraction of red beetroot (Beta vulgaris) peel betalains and phenolics was compared with two extraction methods and solvents. The content of total phenolics in the extracts was determined according to a modification of the Folin-Ciocalteu method and expressed as gallic acid equivalents (GAE). The profiles of extracts were analysed by high-performance liquid chromatography (HPLC). The compounds of beetroot peel extracted with 80% aqueous methanol were characterised from separated fractions using HPLC- diode array detection (HPLC-DAD) and HPLC-electrospray ionisation-mass spectrometry (HPLC-ESI-MS) techniques. The extraction methods and the choice of solvent affected noticeably the content of individual compounds in the extract. The betalains found in beetroot peel extract were vulgaxanthin I, vulgaxanthin II, indicaxanthin, betanin, prebetanin, isobetanin and neobetanin. Also cyclodopa glucoside, N-formylcyclodopa glucoside, glucoside of dihydroxyindol-carboxylic acid, betalamic acid, L-tryptophan, p-coumaric acid, ferulic acid and traces of unidentified flavonoids were detected.

5,5',6,6'-Tetrahydroxy-3,3'-biindolyl from beetroot (Beta vulgaris) peel extract.

A compound of unusual structure was isolated from red beetroot (Beta vulgaris) peel extract and identified as 5,5',6,6'-tetrahydroxy-3,3'-biindolyl based on the combination of NMR and MS studies.

Variation of phlorotannins among three populations of Fucus vesiculosus as revealed by HPLC and colorimetric quantification.

In ecological studies, phlorotannins have conventionally been quantified as a group with similar functionality. Since this group consists of oligo- and polymers, the quantification of their pooled contents alone may not sufficiently describe the variation of these metabolites. Genetic variation, plastic responses to environment, and the ecological functions of separate phlorotannin oligo- and polymers may differ. Two analyses, i.e., the colorimetric Folin-Ciocalteu assay and a normal-phase high-performance liquid chromatographic (HPLC) method were used to study genetic and environmental variation in phlorotannins of the brown alga Fucus vesiculosus (L.). The colorimetric method provides the total phlorotannin content, the latter a profile of 14 separate traces from the phenolic extract that represent an individual or groups of phlorotannins. We reared the algae that originated from three separate populations in a common garden for 3 months under ambient and enriched-nutrient availability and found that they differed in both their total phlorotannin content and in phlorotannin profiles. Some individual traces of the profiles separated the populations more clearly than the colorimetric assay. Although nutrient enrichment decreased total phlorotannin content, it did not show a significant influence on the phlorotannin profile. This implies that plastic responses of compounds other than phlorotannins may interfere with the determination of total phlorotannins. However, the phlorotannin profile and the total content showed genetic variation among local populations of F. vesiculosus; therefore, phlorotannins may respond to natural selection and evolve both quantitatively and qualitatively.

Effects of long-term open-field ozone exposure on leaf phenolics of European silver birch (Betula pendula Roth).

The response of phenolic compounds as a result of long-term low open-field ozone exposure was studied in ozone-sensitive and ozone-tolerant clones of European silver birch (Betula pendula Roth). The saplings were exposed to 1.5-1.6 times the ambient (elevated) ozone and ambient air (as control) over three growing seasons from May 1996 until August 1998. Quantification by modified Folin-Ciocalteau assay showed a 16.2% increase in total phenolics in elevated ozone plants as compared to that in controls and a corresponding 9.9% increase of 10 phenolic compounds quantified by HPLC. Five nonflavonoids and five flavonoids showed 8.4% and 11.4% increases, respectively. The phenolic results indicated slightly higher ozone sensitivity of clone 5 as compared to clone 2. The most ozone-responsive phenolic compounds in clone 2 and clone 5 were (+)-catechin (CT), chlorogenic acid (CGA), 5-p-coumaroylquinic acid (5CQA), 3-p-coumaroylquinic acid (3CQA), myricetin galactopyranoside (MG), quercetin-3-O-glucuronopyranoside (QGR), and quercetin-3-O-arabinofuranoside (QA). Increased phenolic content in ozone-exposed plants was related to impaired growth and accelerated leaf senescence, indicated by enhanced autumn leaf yellowing and lower chlorophyll and Mg content. The change in carbon allocation towards defensive phenolics at the expense of growth was greater in the ozone-sensitive clone as compared to tolerant clone.

Proanthocyanidins of mountain birch leaves: quantification and properties.

Proanthocyanidins (PAs; condensed tannins) are present in mountain birch leaves in soluble and cell wall-bound forms. Crude preparations of soluble PAs were isolated from birch leaves and purified by chromatography on a Sephadex LH-20 column with a yield of about 7% of leaf dry mass. Some chemical characteristics were elucidated with 13C-NMR and HPLC-ECI-MS. Birch leaf PAs were mainly delphinidin type oligo- and polymers with average molecular mass of about 3000. In order to quantify PAs, the method involving heating PA-containing materials in 1-butanol:hydrochloric acid (95:5, v/v), and spectrophotometric determination of the anthocyanidin monomers so formed was modified and optimised. Mature leaves were characterised by a relatively high content of PAs: mean values for soluble and bound PAs were 103 and 40 mg/g dry mass, respectively. In mature leaves the soluble PAs determined the total protein precipitation capacity (PPC) of extracts. In young leaves, the contribution of PAs to the total content of phenolics and the total PPC of tannins was about 20-25% only.

Solvent effects in lipase-catalysed transesterification reactions.

Porcine pancreatic lipase-catalysed transesterifications of 2,2,2-trifluoroethyl butyrate with racemic 2-octanol and 1-phenylethanol have been studied in different organic solvents. Solvent hydrophobicity (log P -1.1 to 3.3) has only a minor effect on the reaction rate. Independently of the solvent used as the reaction medium, both (R)-2-octyl and (R)-1-phenylethyl butyrates were obtained in high optical purity (ee greater than 90%). Candida cylindracea lipase is active only in the most hydrophobic solvents studied.