A High-Content Screen for the Identification of Plant Extracts with Insulin Secretion-Modulating Activity.

Bioactive plant compounds and extracts are of special interest for the development of pharmaceuticals. Here, we describe the screening of more than 1100 aqueous plant extracts and synthetic reference compounds for their ability to stimulate or inhibit insulin secretion. To quantify insulin secretion in living MIN6 ß cells, an insulin-Gaussia luciferase (Ins-GLuc) biosensor was used. Positive hits included extracts from Quillaja saponaria, Anagallis arvensis, Sapindus mukorossi, Gleditsia sinensis and Albizia julibrissin, which were identified as insulin secretion stimulators, whereas extracts of Acacia catechu, Myrtus communis, Actaea spicata L., Vaccinium vitis-idaea and Calendula officinalis were found to exhibit insulin secretion inhibitory properties. Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) were used to characterize several bioactive compounds in the selected plant extracts, and these bioactives were retested for their insulin-modulating properties. Overall, we identified several plant extracts and some of their bioactive compounds that may be used to manipulate pancreatic insulin secretion.

FODMAP Fingerprinting of Bakery Products and Sourdoughs: Quantitative Assessment and Content Reduction through Fermentation.

Fermentable oligo-, di-, and monosaccharides and polyols (FODMAPs) are associated with digestive disorders and with diseases such as irritable bowel syndrome. In this study, we determined the FODMAP contents of bread, bakery products, and flour and assessed the effectiveness of sourdough fermentation for FODMAP reduction. The fermentation products were analyzed to determine the DP 2-7 and DP >7 fructooligosaccharide (FOS) content of rye and wheat sourdoughs. FOSs were reduced by Acetobacter cerevisiae, Acetobacter okinawensis, Fructilactobacillus sanfranciscensis, and Leuconostoc citreum to levels below those in rye (-81%; -97%) and wheat (-90%; -76%) flours. The fermentation temperature influenced the sourdough acetic acid to lactic acid ratios (4:1 at 4 °C; 1:1 at 10 °C). The rye sourdough contained high levels of beneficial arabinose (28.92 g/kg) and mannitol (20.82 g/kg). Our study contributes in-depth knowledge of low-temperature sourdough fermentation in terms of effective FODMAP reduction and concurrent production of desirable fermentation byproducts.

Nutrients, bioactive compounds, and minerals in the juices of 16 varieties of apple (Malus domestica) harvested in Austria: A four-year study investigating putative correlations with weather conditions during ripening.

This study was conducted to examine putative correlations between weather parameters during April-September and the amounts of nutrients, minerals and bioactive compounds in the juices of 16 apple varieties from four harvest years in Lower Austria. For most sugar-parameters, negative correlations were found with the total precipitation (r between -0.42 and -0.64). Conversely, positive correlations were observed with the mean air temperature (r between 0.32 and 0.66), the global radiation (r between 0.32 and 0.61) and the number of tropical days (r between 0.39 and 0.51). The sum of 14 polyphenols (HPLC quantitation) was positively correlated with the mean air temperature and global radiation (rs 0.44 and 0.42). Negative correlations were observed between the global radiation and potassium, magnesium and calcium contents (correlation coefficients -0.49, -0.68 and -0.69). We conclude that increased temperatures and global radiation can be correlated with enhanced sugar synthesis and polyphenol formation.

Acute, reproductive, and developmental toxicity of essential oils assessed with alternative in vitro and in vivo systems.

Essential oils (EOs) have attracted increased interest for different applications such as food preservatives, feed additives and ingredients in cosmetics. Due to their reported variable composition of components, they might be acutely toxic to humans and animals in small amounts. Despite the necessity, rigorous toxicity testing in terms of safety evaluation has not been reported so far, especially using alternatives to animal models. Here, we provide a strategy by use of alternative in vitro (cell cultures) and in vivo (Caenorhabditis elegans, hen's egg test) approaches for detailed investigation of the impact of commonly used rosemary, citrus and eucalyptus essential oil on acute, developmental and reproductive toxicity as well as on mucous membrane irritation. In general, all EOs under study exhibited a comparable impact on measured parameters, with a slightly increased toxic potential of rosemary oil. In vitro cell culture results indicated a concentration-dependent decrease of cell viability for all EOs, with mean IC50 values ranging from 0.08 to 0.17% [v/v]. Similar results were obtained for the C. elegans model when using a sensitized bus-5 mutant strain, with a mean LC50 value of 0.42% [v/v]. In wild-type nematodes, approximately tenfold higher LC50 values were detected. C. elegans development and reproduction was already significantly inhibited at concentrations of 0.5% (wild-type) and 0.1% (bus-5) [v/v] of EO, respectively. Gene expression analysis revealed a significant upregulation of xenobiotic and oxidative stress genes such as cyp-14a3, gst-4, gpx-6 and sod-3. Furthermore, all three EOs under study showed an increased short-time mucous membrane irritation potential, already at 0.5% [v/v] of EO. Finally, GC-MS analysis was performed to quantitate the relative concentration of the most prominent EO compounds. In conclusion, our results demonstrate that EOs can exhibit severe toxic properties, already at low concentrations. Therefore, a detailed toxicological assessment is highly recommended for each EO and single intended application.

Roasted Rye as a Coffee Substitute: Methods for Reducing Acrylamide.

Acrylamide is assumed to be a potential carcinogen, and reference values have therefore been implemented in EU legislation. Thus, the food industry needs to reduce the acrylamide content in consumer products to the lowest possible value. In this study, roasted rye was evaluated for its suitability as a coffee substitution product with respect to its acrylamide content. The influence of process modifiers, free asparagine content, storage, and rye type on the final content of acrylamide was investigated. Changes in carbohydrate composition and brightness caused by the roasting process were described. Sample analysis was conducted via GC-MS and HPLC-CAD. Existing methods were adapted to roasted rye as the sample matrix. CaCl2 and asparaginase treatment as well as pH adjustments prior to roasting did not prove to reduce the acrylamide content. A significantly (* p < 0.027) lower free asparagine content in the raw material resulted in a lower formation of acrylamide in the final product. The acrylamide content significantly decreased (**** p < 0.0001) after 3 (1100 ± 18 µg kg-1) and 6 (490 ± 7 µg kg-1) months of long-term storage. Only samples stored for 6 months (490 ± 7 µg kg-1) met the EU acrylamide content requirements (<500 µg kg-1) for grain-based coffee substitution products.

DNA barcoding for the identification of mold species in bakery plants and products.

Mold identification at the species level in environmental samples is a major challenge. Molecular techniques have been widely used for fungal classification, but as most primers are genus-specific, it is laborious to identify unknown samples. In this study, a PCR-based method for the identification of mold at the species level was developed. Therefore, common sequencing primers and combinations of them, targeting specific DNA regions, were tested. Here we present a combination of eight primer pairs to identify mold within a single PCR run. The approach correctly identified mold of unknown species from samples taken at a local bakery, including Penicillium chrysogenum, Penicillium citrinum, Cladosporium sphaerospermum, Paecilomyces formosus, Rhizopus oryzae and Aspergillus niger. Results obtained from the PCR method were successfully validated by chromatographic mycotoxin and microscopy analysis. Findings highlight DNA barcoding as an appropriate tool for mold identification; however, its efficacy is essentially dependent on DNA quality and primer selection.

Increased Cellular Uptake of Polyunsaturated Fatty Acids and Phytosterols from Natural Micellar Oil.

The transport of hydrophobic compounds to recipient cells is a critical step in nutrient supplementation. Here, we tested the effect of phospholipid-based emulsification on the uptake of hydrophobic compounds into various tissue culture cell lines. In particular, the uptake of ω-3 fatty acids from micellar or nonmicellar algae oil into cell models for enterocytes, epithelial cells, and adipocytes was tested. Micellization of algae oil did not result in adverse effects on cell viability in the target cells. In general, both micellar and nonmicellar oil increased intracellular docosahexaenoic acid (DHA) levels. However, micellar oil was more effective in terms of augmenting the intracellular levels of total polyunsaturated fatty acids (PUFAs) than nonmicellar oil. These effects were rather conserved throughout the cells tested, indicating that fatty acids from micellar oils are enriched by mechanisms independent of lipases or lipid transporters. Importantly, the positive effect of emulsification was not restricted to the uptake of fatty acids. Instead, the uptake of phytosterols from phytogenic oils into target cells also increased after micellization. Taken together, phospholipid-based emulsification is a straightforward, effective, and safe approach to delivering hydrophobic nutrients, such as fatty acids or phytosterols, to a variety of cell types in vitro. It is proposed that this method of emulsification is suitable for the effective supplementation of numerous hydrophobic nutrients.

Hydrophilic Interaction Chromatography Coupled with Charged Aerosol Detection for Simultaneous Quantitation of Carbohydrates, Polyols and Ions in Food and Beverages.

Here, we report an accurate and versatile method for the simultaneous determination of 17 sugars (arabinose, erythrose, fructose, galactose, glucose, isomaltulose, lactose, lyxose, maltose, maltotriose, mannose, raffinose, rhamnose, ribose, sucrose, sorbose and xylose), seven polyols (erythritol, inositol, lactitol, maltitol, mannitol, sorbitol and xylitol), five ions (K+, Br-, Cl-, NO3- and SO42-) and the pseudosaccharide acarbose. For compound separation, hydrophilic interaction chromatography (HILIC) coupled to a corona charged aerosol detector (CAD) was used. The method was validated for linearity, precision, reproducibility, retention factor and optimal injection volume. Standards were measured in the range of 1-1000 mg L-1 and showed good intraday and interday repeatability, as well as precision (relative standard deviation (RSD) < 5%). The LODs and LOQs for the 30 analytes were in the range of 0.032-2.675 mg L-1 and 0.107-8.918 mg L-1, respectively. This method exhibited correlation coefficients of at least R2 > 0.97 for all analytes. The method was tested in 24 food and beverage samples to validate the separation efficiency and sensitivity in natural food matrices and to show the practicability of its use for routine food analysis.