The effect of co-encapsulation of Lactobacillus rhamnosus GG ATCC 53103 with inulin on alginate/chitosan matrix: the viability in fermented soy blend and simulated digestive system

This research verified the ability of Lactobacillus rhamnosus encapsulated with inulin to tolerate the simulated digestive system and their viability in a soy blend. Probiotic encapsulated in alginate-chitosan matrix without inulin presented a better encapsulation efficiency (80.92%) than encapsulation with inulin (57.39%). On the 28th day, the count of probiotics decreased by 3.42 and 1.99 logarithmic cycles of free and encapsulated cells without inulin, respectively. In contrast, the microorganisms encapsulated with inulin showed an increase of 1.26 logs CFU g⁻¹. During gastrointestinal simulation, cell counts decreased by 0.78, 1.55 and 1.95 CFU g⁻¹ logs for encapsulated cells without inulin, free and encapsulated with inulin, respectively. Sensory panellists liked the fermented soy blend with encapsulated lactobacilli, and this result shows the possibility to create new probiotic foods of plant origin. Therefore, the alginate/chitosan matrix can be considered adequate for the encapsulation of L. rhamnosus. The inulin reduces the encapsulation efficiency and increases the cell loss in gastrointestinal simulation. Considering cellular losses, the best option for preparing a fermented soy blend is to use L. rhamnosus encapsulated without inulin.     

A simplified method for the evaluation of fatigue crack front shapes under mode I loading

Two-dimensional elastic or elasto-plastic models dominate the current fatigue crack growth assessment and life prediction procedures for plate components with through-the-thickness cracks. However, as demonstrated in many theoretical and experimental papers, the stress field near the crack tip is always three-dimensional and, as a result, the fatigue crack front is not straight. It is normally curved towards the plate faces. Over the past few years there were a number of very careful numerical studies focusing on the evaluation of fatigue crack front shapes. However, the application of the direct numerical techniques to fatigue phenomena is a very tedious and time consuming process and, sometimes, it is quite ambiguous. In the current paper we develop a simplified method for the evaluation of the front shapes of through-the-thickness fatigue cracks. Further, we validate the developed method against experimental results, investigate the influence of various parameters on the crack front shapes at stable (steady-state) propagation and analyse the differences in the results of fatigue crack growth evaluation obtained with two- and three-dimensional approaches.     

Oxidative Stability and Changes in Chemical Composition of Extra Virgin Olive Oils After Short‐Term Deep‐Frying of French Fries

We aimed at investigating oxidative stability and changes in fatty acid and tocopherol composition of extra virgin olive oil (EVOO) in comparison with refined seed oils during short‐term deep‐frying of French fries, and changes in the composition of the French fries deep‐fried in EVOO. EVOO samples from Spain, Brazil, and Portugal, and refined seed oils of soybean and sunflower were studied. Oil samples were used for deep‐frying of French fries at 180 °C, for up to 75 min of successive frying. Tocopherol and fatty acid composition were determined in fresh and spent vegetable oils. Tocopherol, fatty acid, and volatile composition (by SPME–GC–MS) were also determined in French fries deep‐fried in EVOO. Oil oxidation was monitored by peroxide, acid, and p‐anisidine values, and by Rancimat after deep‐frying. Differential scanning calorimetry (DSC) analysis was used as a proxy of the quality of the spent oils. EVOOs presented the lowest degree of oleic and linoleic acids losses, low formation of free fatty acids and carbonyl compounds, and were highly stable after deep‐frying. In addition, oleic acid, tocopherols, and flavor compounds were transferred from EVOO into the French fries. In conclusion, EVOOs were more stable than refined seed oils during short‐term deep‐frying of French fries and also contributed to enhance the nutritional value, and possibly improve the flavor, of the fries prepared in EVOO.     

Enzymatic de-glycosylation of rutin improves its antioxidant and antiproliferative activities

Bioavailability and biological properties of flavonoid glycosides can be improved after the enzymatic hydrolysis of specific glycosyl groups. In this study, we evaluate the antioxidant and antiproliferative potential of rutin after enzymatic hydrolysis performed by α-l-rhamnosidases (hesperidinase from Penicillium sp. and naringinase from Penicillium decumbens) previously heated at 70 °C for 30 min to inactivate the undesirable β-d-glucosidase activity. The highest in vitro antioxidant activity determined by DPPH radical scavenging was achieved with rutin hydrolyzed by hesperidinase. Rutin was predominantly bioconverted into quercetin-3-glucoside. There was no statistical difference between xanthine oxidase inhibition by rutin before and after hydrolysis. However, in vitro inhibitory activity against ten human tumor cell lines showed that hydrolyzed rutin exerted a more potent antiproliferative effect than quercetin and rutin on various cancer cell lines, specially glioma, and ovarian and breast adenocarcinomas. These results indicate that quercetin-3-glucoside could be a promising functional derivative obtained by rutin hydrolysis.     

Effects of substitutions in the binding surface of an antibody on antigen affinity

The interactions between the Fab and single-chain Fv (scFv) fragments of an antibody (NC10) and its antigen, influenza virus neuraminidase, were analysed in the crystal structures of the Fab-neuraminidase and scFv-neuraminidase complexes. To investigate the contribution to binding made by cavities, salt links and hydrogen bonds in the antibody- antigen interface, 14 single amino acid replacements were made at six contact residues in the scFv fragment by site-directed mutagenesis. The binding affinity of each mutant scFv antibody for neuraminidase was determined with a BIAcore optical biosensor. Four of the mutations resulted in large changes in the free energy of binding to neuraminidase (deltadeltaG > 1 kcal/mol) and together may account for approximately 70% of the free energy of binding. Hence these data support the theory that a small number of residues form the 'functional epitope' and are most important for binding of NC10 to neuraminidase. The salt link between antibody residue (Asp)H56 and (Lys)N432 from neuraminidase was demonstrated to be important for affinity, since substitution of (Asp)H56 with Asn caused a large reduction in the free energy of binding (deltadeltaG = +2.8 kcal/mol). Hydrogen bonds provided by (Tyr)L32 and (Asp)H56 were also important for binding: mutation of (Tyr)L32 to Phe resulted in a significant reduction in binding affinity (deltadeltaG = +1.7 kcal/mol). Disruption of hydrophobic interactions (van der Waals contacts) led to significant reductions in affinity also ((Tyr)H99 to Ala, deltadeltaG = +1.5 kcal/mol; (Leu)L94 to Ala, deltadeltaG > +3.0 kcal/mol). An attempt to increase binding affinity by filling a cavity in the interface with a larger antibody side chain was unsuccessful, as the free energy gained by new antibody-antigen interactions did not compensate for the removal of cavity-bound water molecules.      

Limnological features of Funil Reservoir (R.J., Brazil) and indicator properties of rotifers and cladocerans of the zooplankton community

Funil Reservoir receives inflow from a highly industrialized region and acts as a natural sink to pollutants. Among the consequences of the uncontrolled nutrient loading is the constant presence and periodic heavy blooms of Microcystis aeruginosa . This study verified limnological features and zooplankton assemblage of the Funil Reservoir focusing on the environmental-indicator properties of rotifers and cladocerans. The summer bloom of M. aeruginosa caused reductions in water transparency, nitrate and orthophosphate concentrations and raises in chlorophyll a , pH, dissolved oxygen and chemical oxygen demand values. All zooplankton species presented spatial and temporal variations with the exception of the copepods, which were present in all samples. According to canonical correspondence analysis, ammonium was the variable most related to zooplankton variability and different Rotifera and Cladocera assemblages indicated distinct environment conditions. Rotifer taxa associated with increases of water temperature and chlorophyll a concentration were found with high densities during M. aeruginosa blooms. Species of rotifer and cladocerans are suggested as indicators that can be used to identify different physical and chemical gradients or eutrophic increases in Funil Reservoir.     

Partial oxidation of methane over bimetallic nickel–lanthanide oxides

The study of partial oxidation of methane (POM) over bimetallic nickel–lanthanide oxides was undertaken. Binary intermetallic compounds of the type LnNi (Ln = Pr, Gd, Lu) were used as bimetallic nickel–lanthanide oxides precursors and the products (NiO·Pr₂NiO₄, 2NiO·Gd₂O₃ and 2NiO·Lu₂O₃) were characterized by means of X-ray diffraction, X-ray photoelectron spectroscopy, Raman and temperature programmed reduction. The catalytic activity increases when gadolinium or lutetium replaces praseodymium and the selectivity of the bimetallic nickel–lanthanide oxides is clearly different from that of single metal oxides and/or mechanical mixtures.The existence of an unusual synergism effect between the two metal oxide phases (NiO and Ln₂O₃) that lead to higher conversions of methane and higher selectivities to hydrogen and carbon monoxide correlate also the catalysts stability to deactivation. The activity and selectivity of the gadolinium and lutetium compounds is, under the same conditions, equivalent to that of a platinum commercial catalyst, 5 wt% Pt/Al₂O₃, which stresses the good catalytic performance of this new type of compounds for the production of H₂ and CO (H₂/CO = 2).     

Short communication: effect of tannic acid on composition and ruminal degradability of bermudagrass and alfalfa silages

We measured the effects of the addition of tannic acid on chemical composition and crude protein (CP) ruminal degradability of bermudagrass, cv. coast cross (Cynodon dactylon L.) and alfalfa (Medicago sativa L.) silages with nylon bags incubated in the rumen of two fistulated lactating cows. Silage pH was greater for alfalfa than for bermudagrass. The addition of tannic acid had no effect on silage pH. Ammonia N was greater in alfalfa than in bermudagrass silage. Treated and control silages had similar ammonia N concentrations. The percentage of CP that was solubilized at time 0 was lower for alfalfa than for bermudagrass silage. The addition of tannic acid decreased the portion of CP solubilized at time 0 and increased the potentially degradable fraction of CP. The potentially degradable fraction of CP was greater for alfalfa than for bermudagrass. There was an interaction of species and treatment; the disappearance rate of CP increased with tannic acid treatment of alfalfa but decreased with acid treatment of bermudagrass. The effective degradability of CP was similar for control and treated alfalfa but lower for treated than for control bermudagrass. Tannic acid treatment is effective in decreasing the rapidly soluble fraction of alfalfa and bermudagrass silages, which could be beneficial to the animal because it would decrease the excess of N in the rumen after feeding. Tannic acid treatment decreased CP degradability of bermudagrass silage but had no effect on alfalfa silage, suggesting that tannic acid concentration required to effectively decrease CP degradability differs among forages.     

Expression of human aldehyde dehydrogenase-3 associated with hepatocellular carcinoma: promoter regions and nuclear protein factors related to the expression

Studies in our laboratory have shown that as early as day 8.5 of development, mouse yolk sac cells can generate T cells when placed in a thymic microenvironment. At this stage, yolk sac cells can also differentiate into myeloid cells in vitro. B cell differentiation in vitro was achieved with day 9 yolk sac by providing a bone marrow stromal feeder layer. We have now established endothelial cell lines and clones from yolk sacs of day 8-12 mouse embryos. These vary in their ability to support stem cell maintenance and differentiation. Our principal work has been carried out with day 12 cloned endothelial cell lines. One clone supported the > 100 fold expansion of yolk sac hematopoietic stem cells that subsequently could generate B cells, T cells and myeloid cells both in vitro and in vivo. Preliminary experiments with endothelial cells from younger embryos are also described.