Latex proteins from the plant Calotropis procera are partially digested upon in vitro enzymatic action and are not immunologically detected in fecal material

Soluble proteins from the latex of Calotropis procera (LP) were investigated in vitro and in vivo for digestibility as the latex has previously been shown to produce considerable toxic effects on animals. The latex is also an important biologically active compound that displays antiinflammatory and antidiarrhea properties. The proteins were digested by the action of trypsin, pepsin or chemotrypsin as revealed by gel filtration and SDS-PAGE analysis. Furthermore, the full LP digestion was easily achieved by protease treatment. Rabbit polyclonal antibodies raised against LP failed to detect cross-reactive molecules in fecal material of experimental rats following 35 consecutive days of LP consumption in water. Similar patterns of electrophoresis were observed for the negligible amounts of protein observed in the fecal extracts of control and test animals. No death or toxic effects were observed among animals. Taken together these results suggest that harmful and toxic effects on animals of the latex from C. procera are present in its rubber and low molecular weight fractions rather than its protein content.     

Determination of volatile organic compound patterns characteristic of five unifloral honey by solid-phase microextraction-gas chromatography-mass spectrometry coupled to chemometrics

We report the evaluation of the floral origin of honey by analysis of its volatile organic compounds (VOCs) profile, joined with the use of combined pattern recognition techniques. Honey samples, from five floral origins, were analyzed by headspace solid-phase microextraction-gas chromatography-mass spectrometry, selecting 35 VOCs out of the entire profiles, which were analyzed by hierarchical cluster analysis (HCA), stepwise discriminant analysis (SDA), and K-nearest-neighbor (KNN). Both HCA and SDA were used as exploratory tools to select a group of VOCs representing similitude and differences among studied origins. Thus, six out of 35 VOCs were selected, verifying their discriminating power by KNN, which afforded 93% correct classification. Therefore, we drastically reduced the amount of compounds under consideration but kept a good differentiation between floral origins. Selected compounds were identified as octanal, benzeneacetaldehyde, 1-octanol, 2-methoxyphenol, nonanal, and 2-H-1-benzopyran-2-one. The analysis of VOC profiles, coupled to HCA, SDA, and KNN, provides a feasible alternative to evaluate the botanical source of honey.     

Ecdysteroids of quinoa seeds (Chenopodium quinoa Willd.)

Quinoa (Chenopodium quinoa) is a hardy and nutritious Latin American pseudo-cereal. Studies on the seeds led to the isolation of five ecdysteroids using column chromatography. Their structures were determined as ecdysterone, makisterone A, 24-epi-makisterone A, 24(28)-dehydromakisterone A, and 20,26-dihydroxyecdysone by spectroscopic methods. This study demonstrates that quinoa seeds are a source of ecdysteroids, which were reported to be molting hormones in insects.     

Fingerprints for main varieties of argentinean wines: terroir differentiation by inorganic, organic, and stable isotopic analyses coupled to chemometrics

Our main goal was to investigate if robust chemical fingerprints could be developed for three Argentinean red wines based on organic, inorganic, and isotopic patterns, in relation to the regional soil composition. Soils and wines from three regions (Mendoza, San Juan, and Co?rdoba) and three varieties (Cabernet Sauvignon, Malbec, and Syrah) were collected. The phenolic profile was determined by HPLC-MS/MS and multielemental composition by ICP-MS; (87)Sr/(86)Sr and δ(13)C were determined by TIMS and IRMS, respectively. Chemometrics allowed robust differentiation between regions, wine varieties, and the same variety from different regions. Among phenolic compounds, resveratrol concentration was the most useful marker for wine differentiation, whereas Mg, K/Rb, Ca/Sr, and (87)Sr/(86)Sr were the main inorganic and isotopic parameters selected. Generalized Procrustes analysis (GPA) using two studied matrices (wine and soil) shows consensus between them and clear differences between studied areas. Finally, we applied a canonical correlation analysis, demonstrating significant correlation (r = 0.99; p < 0.001) between soil and wine composition. To our knowledge this is the first report combining independent variables, constructing a fingerprint including elemental composition, isotopic, and polyphenol patterns to differentiate wines, matching part of this fingerprint with the soil provenance.     

Biochemical and spectroscopic characterization of morning glory peroxidase from an invasive and hallucinogenic plant weed Ipomoea carnea

A novel heme peroxidase MGP from the latex of Ipomoea carnea subsp. fistulosa (morning glory) belonging to the Convolvulaceae family was purified to homogeneity using ammonium sulfate precipitation, anion exchange, hydrophobic interaction, and gel filtration chromatography. The enzyme is glycosylated and has a molecular mass of 42.06 kDa (MALDI-TOF) and an isoelectric point of pH 4.3. The enzyme has high yield, broad substrate specificity, and a high stability toward pH, temperature, chaotrophs, and organic solvents. The extinction coefficient (epsilon 280 (1%)) of the enzyme was estimated as 20.56 and it consists of 13 tryptophan, 9 tyrosine, and 8 cysteine residues forming 4 disulfide bridges. There is significant effect of inhibitors targeting S-S bridges (mercaptoethanol, l-cysteine, glutathione), as well as of inhibitors targeting heme (sodium azide and hydroxylamine) on peroxidase activity, whereas inhibition was not observed with ethylmaleinimide due to the absence of reduced cysteine in the enzyme. Polyclonal antibodies against the enzyme have been raised in rabbit, and immunodiffusion suggests that the antigenic determinants of MGP are unique. The N-terminal sequence of MGP (D-E-A-C-I-F-S-A-V-K-E-V-V-D-A) exhibited considerable similarity to the sequence of other known plant peroxidases. Spectroscopic studies (absorbance, fluorescence, and circular dichroism) reveal that MGP has secondary structural features with alpha/beta type with approximately 20% alpha-helicity.     

Evaluation of the antioxidant properties of free and bound phenolic acids from native and malted finger millet (ragi, Eleusine coracana Indaf-15)

Free and bound phenolic acids were isolated from native and malted finger millet (ragi, Eleusine coracana Indaf-15), and their antioxidant properties were evaluated. Protocatechuic, gallic, and caffeic acids were found to be the major free phenolic acids. A 3-fold decrease was observed in protocatechuic acid content, whereas the decrease was marginal in the case of caffeic acid upon 96 h of malting. However, the contents of other free phenolic acids such as gallic, vanillic, coumaric, and ferulic acids increased. Ferulic, caffeic, and coumaric acids were found to be the major bound phenolic acids, and a 2-fold decrease was observed in their contents upon 96 h of malting. The antioxidant activity of a free phenolic acid mixture was found to be higher compared to that of a bound phenolic acid mixture. An increase in antioxidant activity coefficient was observed in the case of free phenolic acids from 770.0 +/- 7.8 to 1686.0 +/- 16.0, whereas the same was decreased from 570.0 +/- 6.0 to 448.0 +/- 4.5 in bound phenolic acids upon 96 h of malting. Therefore, the antioxidant capacity of phenolic acids changes during the malting of ragi.     

Effect of a Bowman-Birk proteinase inhibitor from Phaseolus coccineus on Hypothenemus hampei gut proteinases in vitro

The coffee berry borer, Hypothenemus hampei (Ferrari), is an important devastating coffee pest worldwide. Both trypsin and chymotrypsin enzyme activities from H. hampei larval midgut can be inactivated by proteinaceous enzyme-inhibitors. A serine proteinase inhibitor belonging to the Bowman-Birk class was purified from a wild accession of Phaseolus coccineus L. seeds. The inhibitor (PcBBI1) is a cysteine-rich protein that is heat-stable at alkaline pH. MALDI-TOF/MS analysis showed that PcBBI1 occurs in seeds as a monomer (8689 Da) or dimer (17,378 Da). Using in vitro inhibition assays, it was found that PcBBI1 has a high inhibitory activity against H. hampei trypsin-like enzymes, bovine pancreatic chymotrypsin, and trypsin. According to this, PcBBI1 could be a promising tool to make genetically modified coffee with resistance to coffee berry borer.     

Ultraviolet-C and induced stilbenes control ochratoxigenic Aspergillus in grapes

This study investigated the efficacy of ultraviolet-C (254 nm) and induced stilbenes to inhibit Aspergillus carbonarius and Aspergillus tubingensis and control ochratoxin A production in grapes. In addition, the stilbene synthesis as a response to UV-C treatment and to infection of ochratoxigenic Aspergillus was compared. The initial microbial inactivation by a previously optimized UV-C illumination protocol for increasing trans-resveratrol content in grapes (50 W/m (2), 40 cm, 60 s) was similar on undamaged and damaged grapes, achieving 1.2 and 1.3 log conidia/100 g reductions, respectively. After 5 days of storage at 22 degrees C, UV-C treatment and the stilbenes induced by UV-C inhibited ochratoxigenic Aspergillus growth in undamaged grapes. UV-C elicited the biosynthesis of trans-resveratrol, while microbial infection and tissue damage triggered the biosynthesis of trans-piceid. trans-Resveratrol was not synthesized as a consequence of ochratoxigenic Aspergillus contamination. However, when trans-resveratrol was synthesized by UV-C, it contributed to inhibiting the development of ochratoxin A producing aspergilli. Furthermore, UV-C treatment also contributed to decrease ochratoxin A production by ochratoxigenic aspergilli. Therefore, UV-C is a promising emerging technology either for reducing the potential ochratoxigenic risk in grapes, which is of particular interest to the wine industry, and also for increasing trans-resveratrol content of grapes, which would provide an added value to the wine.     

Protein and oil composition predictions of single soybeans by transmission Raman spectroscopy

The soybean industry requires rapid, accurate, and precise technologies for the analyses of seed/grain constituents. While the current gold standard for nondestructive quantification of economically and nutritionally important soybean components is near-infrared spectroscopy (NIRS), emerging technology may provide viable alternatives and lead to next generation instrumentation for grain compositional analysis. In principle, Raman spectroscopy provides the necessary chemical information to generate models for predicting the concentration of soybean constituents. In this communication, we explore the use of transmission Raman spectroscopy (TRS) for nondestructive soybean measurements. We show that TRS uses the light scattering properties of soybeans to effectively homogenize the heterogeneous bulk of a soybean for representative sampling. Working with over 1000 individual intact soybean seeds, we developed a simple partial least-squares model for predicting oil and protein content nondestructively. We find TRS to have a root-mean-standard error of prediction (RMSEP) of 0.89% for oil measurements and 0.92% for protein measurements. In both calibration and validation sets, the predicative capabilities of the model were similar to the error in the reference methods.     

Inhibition of estrogen-mediated mammary tumorigenesis by blueberry and black raspberry

We previously demonstrated the protective effects of blueberry (BB) and black raspberry (BRB) supplemented at 2.5% dose in an ACI rat mammary tumor model. Here, we assessed a dose-related alteration in tumor indices with diet supplemented with 5% BB or BRB powder. The diet was well tolerated. Tumor palpation from 12 weeks revealed first tumor appearance by 84 days in the control group, that was delayed by 24 and 39 days with the BB and BRB diets, respectively (p = 0.04). Ellagic acid detected in the plasma of rats fed the BRB diet was in the range of 96.6-294.2 ng/mL. While the BB diet showed better efficacy in reducing mammary tissue proliferation and tumor burden, tumor latency was delayed efficiently by BRB. Furthermore, BB was effective in downregulating CYP1A1 expression, while BRB downregulated ERα expression effectively. Distinct anticarcinogenic effects of the two berries correspond to their distinct phytochemical signatures.