Influence of alternative liquid chromatography techniques on the chemical complexity and bioactivity of isolated proanthocyanidin mixtures

Extracts rich in proanthocyanidins, which are implicated in multiple human health benefits, were comparatively separated using alternative separation methods [vacuum or open column liquid chromatography], separation matrices [Toyopearl, Sephadex, or silica gel], and degrees of subfractionation [8 or 12 subfraction series], to evaluate the influence of separation technique on the resolution of the chemical composition and the biological activity of separated proanthocyanidin mixtures in individual subfractions. Bioactivity was assessed using a DNA human topoisomerase II bioassay and structural composition by acid thiolysis (average degree of polymerization, DP) and HPLC-ESI/MS. The amount of parent fraction needed to inhibit 50% of topoisomerase II was 3.38 ng/mL with an average DP of 25.5. A 2(3) factorial analysis revealed that the vacuum and open column strategies for separation, when individually considered, did not yield significantly different results in terms of mass recovery, DP, or bioactivity; however, interactions with other factors such as matrix or subfraction series resulted in distinctive shifts in fraction profiles and biological activity. In general, Sephadex as a matrix permitted elution and separation of discrete, polymerized subfractions with potent inhibition against human topoisomerase II. Sephadex vacuum chromatography, Toyopearl open column chromatography, and Toyopearl vacuum chromatography separation techniques eluted highly polymerized proanthocyanidin mixtures, but the inhibitory bioactivity was attenuated as compared to the parent fraction, whereas Sephadex open column chromatography eluted highly polymerized subfraction mixtures that retained bioactive potential.     

Beta-conglycinin embeds active peptides that inhibit lipid accumulation in 3T3-L1 adipocytes in vitro

Obesity is a worldwide health concern because it is a well-recognized predictor of premature mortality. The objective was to identify soybean varieties that have improved potential to inhibit fat accumulation in adipocytes by testing the effects of soy hydrolysates having a range of protein subunit compositions on lipid accumulation and adiponectin expression in 3T3-L1 adipocytes. The results showed that differences in the protein distribution of 15 soy genotypes led to different potentials for the reduction of fat accumulation. The inhibition of lipid accumulation of soy alcalase hydrolysates in 3T3-L1 adipocytes ranged from 29 to 46%. Soy hydrolysates made from genotypes with 45.3 +/- 3.3% of total protein as beta-conglycinin, on average, showed significantly higher inhibition of lipid accumulation compared to those with 24.7 +/- 1.5% of extracted total protein as beta-conglycinin. Moreover, after in vitro simulated digestion with pepsin-pancreatin of the soy alcalase hydrolysates, 86% of the original activity remained. Adiponectin expression was induced in 3T3-L1 adipocytes treated with 15 soy hydrolysates up to 2.49- and 2.63-fold for high and low molecular weight adiponectin, respectively. The inhibition of lipid accumulation calculated from a partial least squares (PLS) analysis model correlated well with experimental data (R(2) = 0.91). In conclusion, it was feasible to differentiate soy varieties on the basis of the potential of their proteins to reduce fat accumulation using a statistical model and a cell-based assay in vitro. Furthermore, beta-conglycinin embeds more peptides than glycinin subunits that inhibit lipid accumulation and induce adiponectin in 3T3-L1 adipocytes. Therefore, soy ingredients containing beta-conglycinin may be important food components for the control of lipid accumulation in adipose tissue.     

Evolution of arsenate toxicity in nodulated white lupine in a long-term culture

White lupine is an As-resistant legume that is of interest for phytoremediation of As-contaminated soils. To achieve successful phytoremediation, monitoring of the nutritional status of the selected plant species during the entire culture cycle is required to maintain a plant cover with high biomass production. A long-term pot experiment was carried out with nodulated lupine grown on perlite with 10 and 100 microM As concentrations. The reproductive period (from 10 weeks) was the most sensitive phenologic stage of white lupine to long-term As exposure. The 10 microM As treatment increased the uptake and translocation of micronutrients, except for Cu, mainly at flowering with As levels in pods below the statutory limit (1 mg kg (-1) fresh weight). However, the 100 microM As treatment induced significant differences compared to the control. These findings confirm the relatively high resistance of white lupine to arsenate and support the use of this species in phytoremediation and/or revegetation of As-contaminated sites, with special attention on P and Cu nutrition at flowering.     

Alleviation of salt stress by arbuscular mycorrhizal in zucchini plants grown at low and high phosphorus concentration

With the aim of determining whether the arbuscular mycorrhizal (AM) inoculation would give an advantage to overcome salinity problems and if the phosphorus (P) concentration can profoundly influence zucchini (Cucurbita pepo L.) plant responses to AM, a greenhouse experiment was carried out with AM (+AM) and non-AM (−AM). Plants were grown in sand culture with two levels of salinity (1 and 35 mM NaCl, giving electrical conductivity values of 1.8 and 5.0 dS m⁻¹) and P (0.3 and 1 mM P) concentrations. The percentages of marketable yield and shoot biomass reduction caused by salinity were significantly lower in the plants grown at 0.3 mM P, compared to those grown at 1 mM P. However, even at high P concentration, the absolute value of yield and shoot biomass of +AM zucchini plants grown under saline conditions was higher than those grown at low P concentration. The +AM plants under saline conditions had higher leaf chlorophyll content and relative water content than −AM. Mycorrhizal zucchini plants grown under saline conditions had a higher concentration of K and lower Na concentration in leaf tissue compared to −AM plants. The P content of zucchini leaf tissue was similar for +AM and −AM treatments at both low and high P concentrations in the saline nutrient solution. The beneficial effects of AM on zucchini plants could be due to an improvement in water and nutritional status (high K and low Na accumulation).     

Survival of Fusarium circinatum in soil and Pinus radiata needle and branch segments

Survival of Fusarium circinatum in colonized pine needles and wood pieces was measured. Naturally colonized branches and their needles were cut into small pieces and placed in mesh bags on the soil surface at two locations in northern Spain. Pieces were recovered periodically, cultured on a selective medium, and microscopically examined to identify the species. After 507 days, F. circinatum was recovered from 0 to 27% of the wood pieces and from none of the needles. After 858 days, F. circinatum was not recovered from any wood pieces but was found to be present on 1 out of 220 needle pieces analysed. Artificially infested pieces of wood and needles were placed on 5‐mm sieved soil either in plastic boxes at controlled temperature or in mesh bags under field conditions. No survival was recorded after 794 days under field conditions and the decline over time occurred more rapidly in inoculated pieces under field conditions. Soil was also infested with conidia of F. circinatum and survival was estimated. No conidia were recovered after 224 days at 30 °C, although at 20 and 5 °C the respective populations were 20 and 3700 cfu/g soil. Fusarium circinatum was not recovered from 2‐mm‐sieved soil collected under pitch canker‐infected pines. Results indicate that branch segments and needles naturally colonized by F. circinatum will not be a potential source of inoculum, and the fungus in soil is not likely to contribute to reinfection of new plantations after 2 years.     

Lovastatin as a supplement to mitigate rumen methanogenesis: an overviewOA

Methane from enteric fermentation is the gas with the greatest environmental impact emitted by ruminants.Lovastatin(Lv) addition to feedstocks could be a strategy to mitigate rumen methane emissions via decreasing the population of methanogenic archaea(MA). Thus, this paper provides the first overview of the effects of Lv supplementation, focusing on the inhibition of methane production, rumen microbiota, and ruminal fermentation.Results indicated that Lv treatment had a strong anti-methanogenic...