Cytotoxic, antioxidant, and cytoprotective properties of polyphenol-enriched extracts from pecan nutshells in MDA-MB-231 breast cancer cells.

Phenolic compounds present in plants have demonstrated several biological properties such as antioxidant, antitumor, cardioprotective, and antiproliferative. On the other hand, doxorubicin, a chemotherapeutic widely used to treat breast cancer, usually exhibits chronic cardiotoxicity associated with oxidative stress. Therefore, we aimed to study the effects of phenolic compound-enriched extract (PCEE) with doxorubicin in breast cancer. To achieve this, after an SPE-C18 -column purification process of crude extracts obtained from pecan nutshells (Carya illinoinensis), the resulting PCEE was used to evaluate the cytotoxicity and antioxidant properties against the human breast cancer cell line MDA-MB-231 and the normal-hamster ovary cell line CHO-K1. PCEE was selectively cytotoxic against both cell lines, with an IC50 value (≈26.34 mg/L) for MDA-MB-231 lower than that obtained for CHO-K1 (≈55.63 mg/L). As a cytotoxic mechanism, PCEE inhibited cell growth by G2/M cell cycle arrest in MDA-MB-231 cells. Simultaneously, the study of the antioxidant activity showed that PCEE had a cytoprotective effect, evidenced by reduced ROS production in cells with oxidative stress caused by doxorubicin. The results highlight PCEE as a potential antitumor agent, thus revaluing it as an agro-industrial residue.

Intestinal and colonic bioaccessibility of phenolic compounds from fruit smoothies as affected by the thermal processing and the storage conditions.

The effects of the thermal treatment (70 °C for 2 min) and the refrigerated storage (5 °C for 28 d) of a fruit smoothie made of strawberries (40 %), orange juice (20 %), apple (10 %) and banana (10 %) on the microbiological quality, pH and soluble solid contents, color parameters, vitamin C, phenolic compound and antioxidant capacity contents, and on the total in vitro bioaccessibility (intestinal plus colonic) of phenolic compounds were studied. The thermal treatment and the refrigerated storage controlled the microbial load; and did not induce changes in the pH and soluble solid contents of the smoothies. Smoothies were slightly whiter after pasteurization and slightly darker, less red, and less yellow at the end of the refrigerated storage. Ascorbic acid was degraded by the thermal processing and during the storage by up to 50 % at 28 d, and its evolution was modeled by first-order kinetic. Thermal treatment did not produce degradation of the phenolic compounds, nor in the antioxidant capacity of smoothies, and neither elicited change in the intestinal and colonic bioaccessibility of the phenolic compounds. Anthocyanin's retentions declined after 15 days of storage, being around 40% for perlargonidin-3-O-glucoside and pelargonidin-3-O-rutinoside, and non-detectable for cyanindin-3-O-glucoside at day 28. The rest of the phenolic compounds were maintained unchanged during storage. Despite of the degradation of the anthocyanins and other polyphenols over the storage time, their bioaccessibility was not affected during the storage, indicating a possible increase in the extractability and solubility of these compounds from the food matrix by the gastrointestinal system. Anthocyanins were the most abundant phenolic compounds in the smoothies but were those with the lowest total bioaccessibility (≈1.6%). Hence, this work remarks the importance of investigating not only the content and profile of the phenolic compounds as affected by thermal processing and through the storage but also the impact on their bioaccessibility.

Extraction of phenolic compounds from the shells of pecan nuts with cytotoxic activity through apoptosis against the colon cancer cell line HT-29.

The water extraction of phenolic compounds from two varieties ("Mahan" and "Marameck") of pecan nutshells (Carya illinoinensis) without and with sonication, varying the solvent/solid ratio (S), the pH, and the refluxing time (t), was studied. Additionally, the in vitro cytotoxicity and the determination of the cell death mechanism of the extracts against the colon cancer cell line HT-29 were investigated. The content of total phenolic compounds (TPC) of "Marameck" nutshells resulted higher than for the "Mahan" variety, and the pH increase resulted in higher TPC contents for both cultivars. The optimized conditions for TPC extraction without and with sonication resulted: S = 33 ml/g, pH = 12, and t = 9.6 min, and yielded ≈ 70 and 90 mg/g of TPC for "Mahan" and "Marameck" nutshells, respectively. The optimized extracts of pecan nutshells without sonication from both cultivars presented similar cytotoxicity against HT-29 colon cancer cells (IC50  ≈ 50 µg/ml), higher than for sonicated extracts (IC50 ≈ 88 and 138 µg/ml for "Mahan" and "Marameck," respectively). Cell death through apoptosis was the main mechanism of cell death induced by the nutshell extracts. PRACTICAL APPLICATION: The extraction of phenolic compounds (TPC) from the residues of two varieties of pecan nutshells ("Mahan" and "Marameck") was studied. An optimal combination of variables within the pH range that minimizes the solvent-to-solid ratio (S) and the time of refluxing (t), saving at the same time, water and energy, was set up. The phenolic compound extracts obtained from the residues of the pecan nuts exhibit cytotoxic effects against colon cancer cells and could be of interest as an alternative treatment of different types of cancer. Additionally, these extracts may be of importance to the food industry as they can be used as antioxidant agents in food formulation. Also, the high levels of anthocyanidins obtained from the pecan nut extracts after proanthocyanidins' strong acid hydrolysis can be purified and employed as natural red dyes.

Strawberry agro-industrial by-products as a source of bioactive compounds: effect of cultivar on the phenolic profile and the antioxidant capacity.

The post-harvest processing of strawberries generates considerable amounts of by-products that consist of the inedible parts of the fruit (sepal, calyx, stem, and non-marketable portion of the fruit), which is an environmental problem for local producers and industries. This study aimed to revalue these kinds of tissues through identifying and quantifying the genotype influence on the total phenolic content, phenolic profile, and the antioxidant activity of the by-products from three strawberry cultivars: 'Festival' (FE), 'San Andreas ' (SA), and 'Camino Real' (CR). The total phenolic content was determined by the Folin-Ciocalteu method, in-vitro antioxidant activity by the DPPH* radical scavenging method and the phenolic profile by PAD-HPLC. The different genotypes showed significant differences (p < 0.05) in total phenolic content (TPC), FE being the one with the highest TPC (14.97 g of gallic acid equivalents < GAE > /Kg of by-product < R >), followed by SA and CR cultivars. The antioxidant capacity of the SA and FE tissues were similar (p > 0.05) and higher (15.1-16.3 mmol Trolox equivalents < TE > /Kg R) than CR. Eight main phenolic compounds were identified and quantified on the three cultivars. Agrimoniin was the principal polyphenol (0.38-1.56 g/Kg R), and the cultivar FE had the highest concentration. This compound showed the highest correlation coefficient with the antioxidant capacity (R2 0.87; p < 0.001). This study highlighted the impact of the multi-cultivar systems in strawberry production on the bioactive potential and the diversity of secondary metabolites obtained from strawberry agro-industrial by-products at a low cost.

Sous-Vide as a Technique for Preparing Healthy and High-Quality Vegetable and Seafood Products.

Sous-vide is a technique of cooking foods in vacuum bags under strictly controlled temperature, offering improved taste, texture and nutritional values along with extended shelf life as compared to the traditional cooking methods. In addition to other constituents, vegetables and seafood represent important sources of phytochemicals. Thus, by applying sous-vide technology, preservation of such foods can be prolonged with almost full retention of native quality. In this way, sous-vide processing meets customers' growing demand for the production of safer and healthier foods. Considering the industrial points of view, sous-vide technology has proven to be an adequate substitute for traditional cooking methods. Therefore, its application in various aspects of food production has been increasingly researched. Although sous-vide cooking of meats and vegetables is well explored, the challenges remain with seafoods due to the large differences in structure and quality of marine organisms. Cephalopods (e.g., squid, octopus, etc.) are of particular interest, as the changes of their muscular physical structure during processing have to be carefully considered. Based on all the above, this study summarizes the literature review on the recent sous-vide application on vegetable and seafood products in view of production of high-quality and safe foodstuffs.

Changes in the bioactive properties of strawberries caused by the storage in oxygen- and carbon dioxide-enriched atmospheres.

The changes in the antioxidant capacity, anti-inflammatory, and wound healing properties of strawberry fruits as a consequence of the storage in atmospheres enriched in oxygen and carbon dioxide were investigated. Berries were exposed to two different gas compositions: 70% O2 + 20% CO2 and 90% O2 + 10% CO2, and stored for up to 20 days at 5°C. The antioxidant capacity, assessed through DPPH and FRAP methods, decreased around 17% in samples exposed to 70% O2 + 20% CO2 at day 20. However, the antioxidant activity of fruits stored in 90% O2 + 10% CO2 was maintained until day 20 and experienced an increase of around 10% on day 10. Moreover, strawberry stored in 90% O2 + 10% CO2 at days 5-10 showed an improved suppression of the pro-inflammatory genes Cox-2 and iNOS up to 30% higher than samples at day 0 in an in vitro LPS-stimulated RAW 264.7 macrophage culture. In addition, berries exposed to 90% O2 + 10% CO2 at day 10 showed a human dermal fibroblast migration 30% higher than samples at day 0 in an in vitro skin-fibroblast-migration model. Therefore, evidence suggests that strawberry storage in 90% O2 + 10% CO2 can be a promissory alternative to offer fruits with enhanced bioactivity.

Anti-inflammatory and wound healing properties of polyphenolic extracts from strawberry and blackberry fruits.

The polyphenolic profiles by HPLC-TOF-MS of strawberry 'San Andreas' and blackberry 'Black Satin' crude extracts (CE) were analyzed. Anthocyanin-enriched fractions (AEFs) and proanthocyanidin-enriched fractions (PEFs) were prepared, and all samples were probed for in vitro anti-inflammatory and wound healing effects in a LPS-stimulated RAW 264.7 macrophage model and in a skin fibroblast migration and proliferation assay, respectively. Blackberry samples exhibited higher ROS reduction than strawberry's (up to 50% ROS suppression). Berries CEs exhibited 20% inhibition in Cox-2 gene expression, while AEFs and PEFs were inactive at the same concentration. Strawberry AEF and PEF were more active against IL-1ß and IL-6 gene expressions than the similar fractions from blackberry, where PEF was more active than AEF (75% suppression by strawberry PEF). Moreover, berry PEFs were the active polyphenol fraction against iNOS gene expression (50% and 65% gen suppression by strawberry and blackberry PEF, respectively), mirroring results of NO synthesis suppression. The cell migration potential of berry polyphenolics was associated with anthocyanins. AEFs showed fibroblast migration around 50% of that registered for the positive control. Results obtained in this work highlight the anti-inflammatory properties of berry polyphenolics, especially due to proanthocyanidins. Moreover, promising results were obtained about the effects of berry anthocyanins on wound healing.

Optimization of strawberry disinfection by fogging of a mixture of peracetic acid and hydrogen peroxide based on microbial reduction, color and phytochemicals retention.

The fogging of strawberries using a environmentally friendly sanitizer mixture of peracetic acid (5%) and hydrogen peroxide (20%) was performed in a model chamber and modeled as a function of the concentration (3.4, 20.0, 60.0, 100.0 and 116.6 µL sanitizer L(-) (1) air chamber) and the treatment time (5.7, 15.0, 37.5, 60.0 and 69.3 min). The sanitizer fogging was adequate for reducing total mesophilic microbial and yeasts and moulds counts of fruits until seven days of storage at 2℃. However, sanitizer oxidant properties adversely affected the content of total anthocyanins, total phenolics, vitamin C, and antioxidant capacity to various degrees, with some deleterious changes in the fruits color, depending on the fogging conditions. A multiple numeric response optimization was developed based on 2.0 log microbiological reduction, maximum phytochemicals and antioxidant capacity retentions, with no changes in the fruits color, being the optimal fogging conditions achieved: 10.1 µL sanitizer L(-1) air chamber and 29.6 min. The fogging of strawberries at these conditions may represent a promising postharvest treatment option for extending their shelf-life without affecting their sensory quality and bioactive properties.

Bioactive Compounds and Antioxidant Capacity of Camarosa and Selva Strawberries (Fragaria x ananassa Duch.).

Strawberries represent an important source of bioactive compounds due to their vitamin C and phenolic compound levels, which present high antioxidant effects, beneficial for the maintenance of consumer's health. Argentina is the second largest strawberry producer in The Common Market of the Southern Cone (MERCOSUR), covering the main export destinations of Argentinian strawberries, i.e., Canada, United States, and European Union. Information about the bioactive compound occurrence and antioxidant capacity of these fruits is scarce or not available. Health related compounds of strawberry cultivars (Camarosa and Selva) from different zones of Argentina were investigated. Vitamin C content was in the same range for both studied cultivars. However, Camarosa strawberries, which are the most cultivated, consumed, and exported berries in Argentina, presented higher total phenolic and anthocyanins content, and consequently better in vitro antioxidant capacity. Moreover, there were differences in the occurrence and concentration in the phenolic compound profiles for both cultivars. Camarosa cultivar presented higher content of anthocyanidins, and Selva showed higher total ellagic acid content. The research shows that Argentina's strawberries are an interesting source of bioactive compounds comparable to those in other parts of the world.