Novel extracellular polymeric substances produced by Cupriavidus necator IPT 027 grown on glucose and crude glycerol originated from biodiesel

The aim of this study was to evaluate the production and properties of novel extracellular polymeric substances (EPS) from glucose (EPS 1) and crude glycerol (CG) from castor beans oil (EPS 2). Cupriavidus necator IPT 027 cultivated with glucose and CG (35 °C, 150 rpm, 72 hr, pH 7, shaker) produced approximately 1.60 ± 0.01 and 2.83 ± 0.02 g l^?1 EPS, respectively, with high‐molecular weight (3.89 × 10⁵ and 1.89 × 10⁷ Da) and constituted of different functional groups such as uronic acid, monosaccharides (glucose, mannose, arabinose, and fucose) and primary amine group. The composition of the fermentable substrate influenced the melting temperature (221.11 and 230.18 °C), crystallinity (34.36% and 37.11%), degradation temperature (255.06 and 296.62 °C) and morphology. EPS showed pseudoplastic non‐Newtonian fluid behavior in the aqueous solutions, presenting potential applicability biotechnological and industrial mainly in food industry as emulsifiers and biosurfactants. This is the first study on the production and characterization of EPS obtained by C. necator IPT 027 in culture with glucose and CG. Copyright © 2016 John Wiley & Sons, Ltd.     

Cytotoxicity of Frutalin on Distinct Cancer Cells Is Independent of Its Glycosylation

Frutalin is a plant lectin with beneficial immunobiological action, although the access to its active form is still restricted. Moreover, there is a knowledge gap on isoform activity and glycosylation impact on its bioactivity, and recombinant production protocols were seen as ineffective. Here, a simpler and faster production and purification protocol was developed, attaining a yield of purified frutalin 3.3-fold higher than that obtained previously. Hemagglutination assays confirmed that this frutalin isoform could not agglutinate rabbit erythrocytes, while maintaining the native tetrameric structure, as indicated by DLS analysis, and strong interaction with methyl-alpha-galactose, in fluorescence spectroscopy studies. The cytotoxicity of the recombinant frutalin isoform was shown in a broad panel of human cancer cells: colon (HCT116), melanoma (A375), triple-negative breast cancer (MDA-MB-231), and ovarian (IGROV-1). Treatment with 8.5–11.8 μM TrxFTL reduced proliferation of all cancer cells to half in 48 h. This anti-proliferative effect encompasses the p53 pathway since it was significantly reduced in p53-null colon cancer cells (HCT116 p53^−/−; GI₅₀ of 25.0 ± 3.0 μM), when compared to the isogenic p53-positive cells (HCT116 p53^+/+; GI₅₀ of 8.7 ± 1.8 μM; p < 0.002). This recombinantly produced frutalin isoform has relevant cytotoxic effect and its biological activity is not dependent on glycosylation. The developed E. coli production and purification protocol generates high yield of non-glycosylated frutalin isoform with potent cytotoxic activity, enabling the development of novel anticancer p53-targeting therapies.     

Antiprotozoal and Antibacterial Activity of Ravenelin,a Xanthone Isolated from the Endophytic Fungus Exserohilum rostratum

The natural compound ravenelin was isolated from the biomass extracts of Exserohilum rostratum fungus, and its antimicrobial, antiplasmodial, and trypanocidal activities were evaluated. Ravenelin was isolated by column chromatography and HPLC and identified by NMR and MS. The susceptibility of Gram-positive and Gram-negative bacteria strains to ravenelin was determined by microbroth dilution assay. Cytotoxicity was evaluated in hepatocarcinoma cells (HepG2) and BALB/c peritoneal macrophages by using MTT. SYBR Green I-based assay was used in the asexual stages of Plasmodium falciparum. Trypanocidal activity was tested against the epimastigote and intracellular amastigote forms of Trypanosoma cruzi. Ravenelin was active against Gram-positive bacteria strains, with emphasis on Bacillus subtilis (MIC value of 7.5 µM). Ravenelin’s antiparasitic activities were assessed against both the epimastigote (IC₅₀ value of 5 ± 1 µM) and the intracellular amastigote forms of T. cruzi (IC₅₀ value of 9 ± 2 µM), as well as against P. falciparum (IC₅₀ value of 3.4 ± 0.4 µM). Ravenelin showed low cytotoxic effects on both HepG2 (CC₅₀ > 50 µM) and peritoneal macrophage (CC₅₀ = 185 ± 1 µM) cells with attractive selectivity for the parasites (SI values > 15). These findings indicate that ravenelin is a natural compound with both antibacterial and antiparasitic activities, and considerable selectivity indexes. Therefore, ravenelin is an attractive candidate for hit-to-lead development.     

Classical Food Quality Attributes and the Metabolic Profile of Cambuci,a Native Brazilian Atlantic Rainforest Fruit

The cambuci is a native Brazilian fruit from the Atlantic Forest biome. A soft and astringent pulp, a green color, and a sweet aroma are its main characteristics. Classical food quality attributes (fresh fruit mass, fruit height, diameters, total soluble solid, titratable acidity, and ratio) and the metabolic profile from ten accessions from three different locations were analyzed herein by analytical methods (refractometry and neutralization titration) and nuclear magnetic resonance spectroscopy. Concerning sugar content, sucrose was the predominant compound, with glucose and fructose alternating in second, depending on the accession. Citric acid was the most relevant acid, followed by shikimic and quinic acids in quite variable amounts. These three main acids vary in amounts for each accession. Ascorbic acid content emerges as an important quality attribute and makes this fruit nutritionally attractive, due to values comparable to those contained in citric fruits. The main amino acids identified in cambuci were glutamic acid individually or in comprising the tripeptide glutathione (glutamic acid, cysteine, glycine). The quality diversity of the evaluated accessions suggests the potentiality of cambuci use in future breeding programs.     

Cosmetic Formulations with Melaleuca alternifolia Essential Oil for the Improvement of Photoaged Skin: A Double-Blind,Randomized, Placebo-Controlled Clinical Study

This aim of this study was to evaluate the penetration depth, antioxidant capacity and the clinical efficacy of Melaleuca alternifolia pure essential oil and in a nanoemulsion to prevent skin photoaging. For this, 2% of pure essential oil or 2% of this essential oil in a nanoemulsion were vehiculated in a formulation. The skin penetration was evaluated using confocal Raman microspectroscopy. The radical protection factor was evaluated using electron paramagnetic resonance spectroscopy. For a clinical study, 40 male participants, aged 18–28 years, were enrolled, being divided into three groups: vehicle formulation, M. alternifolia pure essential oil and M. alternifolia Nanoemulsion. All the participants also received a sunscreen SPF 50 to use during the day. Before and after 90 days of study, skin hydrolipidics and morphological characteristics were performed by skin imaging and biophysical techniques. The nanoemulsion presented a lower antioxidant capacity and a higher penetration through the stratum corneum, reaching the viable epidermis, improving the stratum granulosum morphology. The groups presented an increase in the papillary depth, improving in the dermis echogenicity and the collagen fibers. Melaleuca alternifolia essential provides the potential to improve photoaged skin, being the application of nanoemulsion able to reach deeper skin layers.     

GRASP with path relinking for the orienteering problem

In this paper, we address an optimization problem resulting from the combination of the well-known travelling salesman and knapsack problems. In particular, we target the orienteering problem, originated in the context of sport, which consists of maximizing the total score associated with the vertices visited in a path within the available time. The problem, also known as the selective travelling salesman problem, is NP-hard and can be formulated as an integer linear program. Since the 1980s, several solution methods for this problem have been developed and applied to a variety of fields, particularly in routing and tourism. We propose a heuristic method—based on the Greedy Randomized Adaptive Search Procedure (GRASP) and the Path Relinking methodologies—for finding approximate solutions to this optimization problem. We explore different constructive methods and combine two neighbourhoods in the local search of GRASP. Our experimentation with 196 previously reported instances shows that the proposed procedure obtains high-quality solutions employing short computing times.     

Cohen and multiple Cohen strongly summing multilinear operators

Considering the successful theory of multiple summing multilinear operators as a prototype, we introduce the classes of multiple Cohen strongly p-summing multilinear operators and polynomials. The adequacy of these classes under the viewpoint of the theory of multilinear and polynomial ideals and holomorphy types is discussed in detail.     

Quadrature formulas for the Laplace and Mellin transforms

A discrete Laplace transform and its inversion formula are obtained by using a quadrature of the integral Fourier transform which is given in terms of Hermite polynomials and its zeros. This approach yields a convergent discrete formula for the two-sided Laplace transform if the function to be transformed falls off rapidly to zero and satisfies given conditions of integrability, achieving convergence also for singular functions. The inversion formula becomes a quadrature formula for the Bromwich integral. The use of asymptotic formulae yields an algorithm to compute the discrete Laplace transform by using only exponentials.     

Strategies for spectrum slicing based on restarted Lanczos methods

In the context of symmetric-definite generalized eigenvalue problems, it is often required to compute all eigenvalues contained in a prescribed interval. For large-scale problems, the method of choice is the so-called spectrum slicing technique: a shift-and-invert Lanczos method combined with a dynamic shift selection that sweeps the interval in a smart way. This kind of strategies were proposed initially in the context of unrestarted Lanczos methods, back in the 1990’s. We propose variations that try to incorporate recent developments in the field of Krylov methods, including thick restarting in the Lanczos solver and a rational Krylov update when moving from one shift to the next. We discuss a parallel implementation in the SLEPc library and provide performance results.     

Strategic oscillation for the capacitated hub location problem with modular links

The capacitated single assignment hub location problem with modular link capacities is a variant of the classical hub location problem in which the cost of using edges is not linear but stepwise, and the hubs are restricted in terms of transit capacity rather than in the incoming traffic. We propose a metaheuristic algorithm based on strategic oscillation, a methodology originally introduced in the context of tabu search. Our method incorporates several designs for constructive and destructive algorithms, together with associated local search procedures, to balance diversification and intensification for an efficient search. Computational results on a large set of instances show that, in contrast to exact methods that can only solve small instances optimally, our metaheuristic is able to find high-quality solutions on larger instances in short computing times. In addition, the new method, which joins tabu search strategies with strategic oscillation, outperforms the previous tabu search implementation.