Comparative bioactivity assessment of bixin pigment and associated phytochemicals extracted from annatto seeds using conventional and green solvents.

Nutraceuticals, that include food ingredients and bioactives from natural products, confer physiological health benefits and protection against chronic diseases. Annatto is a tropical shrub grown in Central and South America and parts of India. Its seeds are rich in the edible carotenoid-derived apocarotenoid pigment, bixin, which is used as a natural colorant in food, textiles, and cosmetics, and is now gaining attention for its potential health-promoting attributes. Here, we compared a green solvent (ethyl lactate) based extraction of bixin and associated metabolites in annatto seeds (crushed and seed coat) with two other conventional solvents (acetone and acid-base). Bixin was characterized in the extracts using UV-visible- and FTIR-spectroscopy and thin-layer chromatography. The bixin-containing solvent extracts were then profiled for other co-existing metabolites using GC-MS analysis, which were found to be sesquiterpenes, terpenes, terpenoids, phytosterols, and tocotrienols. Their bioactivity was evaluated based on antioxidant and wound-healing efficacies and compared with pure bixin, using NIH-3T3 fibroblast cells in-vitro. Pure bixin, as well as the annatto solvent extracts, showed strong antioxidant and wound healing properties, wherein pure bixin and green solvent extract (ethyl lactate coat) exhibited higher levels of antioxidant activity, achieving 46.00% and 44.60% reduction in MDA levels, respectively, as well as enhanced wound-healing activity, with 54.09% and 53.60% wound closure within 24 h. The green solvent extracts of annatto seeds revealed: (a) differential bioactive profiles in annatto seeds (crushed and seed coat) in comparison with other solvents, and (b) strong antioxidant and wound healing properties. Thus, ethyl lactate extraction shows strong potential for sustainable environmental friendly production of functional foods/nutraceuticals from annatto seeds.

Green Methods to Recover Bioactive Compounds from Food Industry Waste: A Sustainable Practice from the Perspective of the Circular Economy.

The worrying and constant increase in the quantities of food and beverage industry by-products and wastes is one of the main factors contributing to global environmental pollution. Since this is a direct consequence of continuous population growth, it is imperative to reduce waste production and keep it under control. Re-purposing agro-industrial wastes, giving them new life and new directions of use, is a good first step in this direction, and, in global food production, vegetables and fruits account for a significant percentage. In this paper, brewery waste, cocoa bean shells, banana and citrus peels and pineapple wastes are examined. These are sources of bioactive molecules such as polyphenols, whose regular intake in the human diet is related to the prevention of various diseases linked to oxidative stress. In order to recover such bioactive compounds using more sustainable methods than conventional extraction, innovative solutions have been evaluated in the past decades. Of particular interest is the use of deep eutectic solvents (DESs) and compressed solvents, associated with green techniques such as microwave-assisted extraction (MAE), ultrasonic-assisted extraction (UAE), pressurized liquid extraction (PLE) and pulsed-electric-field-assisted extraction (PEF). These novel techniques are gaining importance because, in most cases, they allow for optimizing the extraction yield, quality, costs and time.

Isoquinoline-based intrinsic fluorescence assessment of erythropoiesis-stimulating agent, Roxadustat (FG-4592), in tablets: applications to content uniformity and human plasma evaluation.

In the present study, a first validated and green spectrofluorimetric approach for its assessment and evaluation in different matrices was investigated. After using an excitation wavelength of 345 nm, Roxadustat (ROX) demonstrates a highly native fluorescence at an emission of 410 nm. The influences of experimental factors such as pH, diluting solvents, and different organized media were tested, and the most appropriate solvent choice was ethanol. It was confirmed that there was a linear relationship between the concentration of ROX and the relative fluorescence intensity in the range 60.0-1000.0 ng ml-1, with the limit of detection and limit of quantitation, respectively, being 17.0 and 53.0 ng ml-1. The mean recoveries % [±standard deviation (SD), n = 5] for pharmaceutical preparations were 100.11% ± 2.24%, whereas for plasma samples, they were 100.08 ± 1.08% (±SD, n = 5). The results obtained after the application of four greenness criteria, Analytical Eco-Scale metric, NEMI, GAPI, and AGREE metric, confirmed its eco-friendliness. In addition, the whiteness meter (RGB12) confirmed its level of sustainability. The International Council for Harmonisation (ICH) criteria were used to verify the developed method through the study in both spiked plasma samples and content uniformity evaluation. An appropriate standard for various applications in industry and quality control laboratories was developed.

Do Ionic Liquids Exhibit the Required Characteristics to Dissolve, Extract, Stabilize, and Purify Proteins? Past-Present-Future Assessment.

Proteins are highly labile molecules, thus requiring the presence of appropriate solvents and excipients in their liquid milieu to keep their stability and biological activity. In this field, ionic liquids (ILs) have gained momentum in the past years, with a relevant number of works reporting their successful use to dissolve, stabilize, extract, and purify proteins. Different approaches in protein-IL systems have been reported, namely, proteins dissolved in (i) neat ILs, (ii) ILs as co-solvents, (iii) ILs as adjuvants, (iv) ILs as surfactants, (v) ILs as phase-forming components of aqueous biphasic systems, and (vi) IL-polymer-protein/peptide conjugates. Herein, we critically analyze the works published to date and provide a comprehensive understanding of the IL-protein interactions affecting the stability, conformational alteration, unfolding, misfolding, and refolding of proteins while providing directions for future studies in view of imminent applications. Overall, it has been found that the stability or purification of proteins by ILs is bispecific and depends on the structure of both the IL and the protein. The most promising IL-protein systems are identified, which is valuable when foreseeing market applications of ILs, e.g., in "protein packaging" and "detergent applications". Future directions and other possibilities of IL-protein systems in light-harvesting and biotechnology/biomedical applications are discussed.

Efficient Sampling of Cavity Hydration in Proteins with Nonequilibrium Grand Canonical Monte Carlo and Polarizable Force Fields.

Prediction of the hydration levels of protein cavities and active sites is important to both mechanistic analysis and ligand design. Due to the unique microscopic environment of these buried water molecules, a polarizable model is expected to be crucial for an accurate treatment of protein internal hydration in simulations. Here we adapt a nonequilibrium candidate Monte Carlo approach for conducting grand canonical Monte Carlo simulations with the Drude polarizable force field. The GPU implementation enables the efficient sampling of internal cavity hydration levels in biomolecular systems. We also develop an enhanced sampling approach referred to as B-walking, which satisfies detailed balance and readily combines with grand canonical integration to efficiently calculate quantitative binding free energies of water to protein cavities. Applications of these developments are illustrated in a solvent box and the polar ligand binding site in trypsin. Our simulation results show that including electronic polarization leads to a modest but clear improvement in the description of water position and occupancy compared to the crystal structure. The B-walking approach enhances the range of water sampling in different chemical potential windows and thus improves the accuracy of water binding free energy calculations.

BAY 41-2272 activates host defence against local and disseminated Candida albicans infections

In our previous study, we have found that 5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]-pyrimidin-4-ylamine (BAY 41-2272), a guanylate cyclase agonist, activates human monocytes and the THP-1 cell line to produce the superoxide anion, increasing in vitro microbicidal activity, suggesting that this drug can be used to modulate immune functioning in primary immunodeficiency patients. In the present work, we investigated the potential of the in vivo administration of BAY 41-2272 for the treatment of Candida albicans and Staphylococcus aureus infections introduced via intraperitoneal and subcutaneous inoculation. We found that intraperitoneal treatment with BAY 41-2272 markedly increased macrophage-dependent cell influx to the peritoneum in addition to macrophage functions, such as spreading, zymosan particle phagocytosis and nitric oxide and phorbol myristate acetate-stimulated hydrogen peroxide production. Treatment with BAY 41-2272 was highly effective in reducing the death rate due to intraperitoneal inoculation of C. albicans, but not S. aureus. However, we found that in vitro stimulation of peritoneal macrophages with BAY 41-2272 markedly increased microbicidal activities against both pathogens. Our results show that the prevention of death by the treatment of C. albicans-infected mice with BAY 41-2272 might occur primarily by the modulation of the host immune response through macrophage activation. .

In vitro assessment of solvent evaporation from commercial adhesive systems compared to experimental systems

Solvents should be properly evaporated after application to dental substrates. The aim of this study was to assess the evaporation of commercial, experimental and neat solvents. The tested null hypotheses were that there are no differences in solvent evaporation regardless of its formulation and over time. Evaporation from commercial adhesive systems (Scotchbond Multipurpose Primer, Scotchbond Multipurpose Adhesive, Prime & Bond NT, Multi Bond, Excite, Single Bond 2, Adhese Primer, Adhese Bond, Xeno III A and Xeno III B) and experimental primers (35 percent HEMA plus 65 percent acetone or ethanol or water v/v) were compared to neat solvents (acetone, ethanol and water). Samples (10 µL) of these products were dripped into glass containers placed on a digital precision balance. Evaporation was assessed at 0, 5, 10, 15, 30, 60, 120, 300 and 600 s times to calculate mass loss. Data were analyzed statistically by ANOVA and Bonferroni's correction (a=0.05). Acetone-based products exhibited a remarkable capacity to evaporate spontaneously over time. Neat acetone evaporated significantly more than the HEMA-mixtures and the commercial formulations (p<0.05). The incorporation of monomers and other ingredients in the commercial formulations seem to reduce the evaporation capacity. Solvent evaporation was time and material-dependent.

O solvente deve ser adequadamente evaporado após aplicação ao substratos dentários. O objetivo deste estudo foi avaliar a evaporação de formulações comerciais, primers experimentais e solventes puros. As hipóteses nulas testadas foram de que não há diferenças da quantidade evaporada independentemente do material e tempo. Evaporação dos sistemas adesivos comerciais (Scotchbond multipurpose primer, Scotchbond multipurpose adhesive, Prime & Bond NT, Multi Bond, Excite, Single Bond 2, Adhese Primer, Adhese Bond, Xeno III A e Xeno III B) e primers experimentais (35 por cento HEMA associado com 65 por cento acetona, etanol ou água v/v) foram comparadas a solventes puros (acetona, etanol e água). Amostras (10 µL) de cada produto foram dispensadas em balança de precisão digital. As massas nos tempos 0, 5, 10, 15, 30, 60, 120, 300 e 600 s foram registradas. Os dados foram analisados estatisticamente por ANOVA e Bonferroni (a=0,05). Produtos a base de acetona exibiram maior capacidade de evaporação espontânea ao longo do tempo. Acetona pura evaporou significantemente mais que as misturas de HEMA e formulações comerciais (p<0,05). A incorporação de monômeros e outros ingredientes nas formulações comerciais reduzem a capacidade de evaporação. A evaporação é dependente do produto e do tempo.