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Synergies between compounds in food ingredients have been commonly obtained by the mixing of extracts produced individually. A different approach, based on the coextraction of plants and a new theoretical framework to identify synergistic and antagonistic effects in the coextracts, has been proposed in this study for the first time. The Pressurized Liquid Extraction (PLE) of rosemary and marjoram with parsley at several plant ratios was carried out and the antioxidant activity (ABTS and cellular assays) and phenolic compounds of the extracts were evaluated. Synergistic effects on the antioxidant activity of the extracts were identified from all the plant combinations. The HPLC-PAD analysis of the samples did not show a relation between the concentration of the main phenolic compounds in the coextracts and the observed antioxidant synergistic effects. The proposed theoretical framework may be successfully used for the determination of synergies in the bioactivity of extracts obtained by coextraction.

Pressurized liquid extraction (PLE) and ultrasound assisted extraction (UAE) were applied to extract phenolic antioxidants from defatted Brazil nut skin (DBNS) industrial by-product. Water/alcohol solvents (ethanol and methanol) (0 - 50 % v/v water) were used at temperatures of 50 - 200 °C in PLE and 50 °C was set in UAE. The effect on extraction yield, total phenolic compounds content (TPC), antioxidant capacity and phenolic characterization was determined. The temperature and water/alcohol mixture affected the dielectric constant, and this in turn influenced the yield, TPC, antioxidant activity and phenolic composition. In PLE, at 200 °C and 50 % (v/v) water/alcohol (ethanol and methanol) the highest yields (33.8 and 38.4 %, respectively), TPC (4673 and 5435 mg GAE/100 g DBNS) and antioxidant capacity (397 and 502 µmol TE/g DBNS) were obtained. Furthermore, at 50 °C, PLE had better extraction efficiency than UAE. Characterization of the extracts indicated that at 50 - 100 °C the extraction of protocatechuic acid and ellagic acid derivative were predominant, while at 150 - 200 °C catechin and vanillin derivatives stand out. In general, hydroxybenzoic acids and flavonoids were the most abundant phenolics, and the addition of 25 % v/v water to ethanol resulted in extracts with their highest concentration. Then, Brazil nut skin is an interesting phenolic-rich source with potential use to produce bioactive ingredients.

The effect of high-pressure homogenization (HPH) on the inactivation of Escherichia coli and the stability of the quality properties of Brazil nut beverages were studied. E. coli was used as target microorganism to validate the HPH process (pressures from 50 to 180 MPa and inlet temperatures (Ti) from 25 to 75 °C). Cold storage (5 °C) for 21 days was conducted to establish the shelf-life of BN beverages, in terms of their microbiological, physical, physicochemical, and sensorial stability. HPH-treated samples were compared to pasteurized BN beverages (63 °C for 20 min). The combination of Ti and the pressure of the HPH process (50 to 150 MPa/75 °C and 180 MPa/25 °C) had a significant effect on E. coli inactivation (8.2 log CFU/mL). During storage at 5 °C, the growth of mesophilic aerobes in processed BN beverages was controlled by the HPH process. Oxidative stability (TBAR assay) and physicochemical properties (pH, acidity, and °Brix) were evaluated during cold storage, showing good stability. Additionally, HPH-treated beverages showed a reduction in their particle size and the formation of more stable protein aggregates, which favored the beverages’ whiteness (color). The HPH process could be an alternative to pasteurization to obtain Brazil nut beverages with an acceptable microbiological shelf life (≥21 days at 5 °C) and high-quality characteristics without the use of any additives.

Synsepalum dulcificum (Richardella dulcifica) is a berry fruit from West Africa with the ability to convert the sour taste into a sweet taste, and for this reason, the fruit is also known as the “miracle berry” (MB). The red and bright berry is rich in terpenoids. The fruit’s pulp and skin contain mainly phenolic compounds and flavonoids, which correlate with their antioxidant activity. Different polar extracts have been described to inhibit cell proliferation and transformation of cancer cell lines in vitro. In addition, MB has been shown to ameliorate insulin resistance in a preclinical model of diabetes induced by a chow diet enriched in fructose. Herein, we have compared the biological activities of three supercritical extracts obtained from the seed—a subproduct of the fruit—and one supercritical extract obtained from the pulp and the skin of MB. The four extracts have been characterized in terms of total polyphenols content. Moreover, the antioxidant, anti-inflammatory, hypo-lipidemic, and inhibition of colorectal cancer cell bioenergetics have been compared. Non-polar supercritical extracts from the seed are the ones with the highest effects on the inhibition of bioenergetic of colorectal (CRC) cancer cells. At the molecular level, the effects on cell bioenergetics seems to be related to the inhibition of main drivers of the de novo lipogenesis, such as the sterol regulatory element binding transcription factor (SREBF1) and downstream molecular targets fatty acid synthase (FASN) and stearoyl coenzyme desaturase 1 (SCD1). As metabolic reprograming is considered as one of the hallmarks of cancer, natural extracts from plants may provide complementary approaches in the treatment of cancer. Herein, for the first time, supercritical extracts from MB have been obtained, where the seed, a by-product of the fruit, seems to be rich in antitumor bioactive compounds. Based on these results, supercritical extracts from the seed merit further research to be proposed as co

The supercritical defatting of H. illucens was scaled up at 450 bar and 60 °C from a 270 cm3 extraction cell to a vessel five times larger. Then, eight different H. illucens larvae batches, with variable content of oil (16.80–29.17% w/w) and moisture (4.45–15.95% w/w) were defatted. The effect of these parameters on yield and oil composition was analyzed. The presence of moisture in the larvae batch, in the range of the values studied, had no negative effect on the oil recovery efficiency, which was mainly determined by the initial content of oil in the larvae samples. Furthermore, no differences were determined in the fatty acid profile of the oils recovered, which were rich in saturated fatty acids, mainly lauric acid (ca. 50% w/w). Minor lipids, such as squalene and phytosterols, were determined in all the oil samples. The moisture content in the oils extracted was in the range of 0.118–1.706% w/w. Therefore, some samples exceeded the limits recommended for volatile matter in edible fats and oils (0.2%, including moisture). Yet, concerning the oil peroxide index, values were much lower than those corresponding to the oil extracted using hexane.

Borututu (Cochlospermum angolense Welw.) roots have been described as a rich source of phenolic compounds. Despite the potential of this plant for the production of bioactive extracts, studies reported until now have been scarce, and they have been based on the use of inefficient conventional extraction techniques. In this study, pressurized liquid extraction (PLE) was investigated for the production of borututu root extracts. Different temperatures (50–200 °C) and solvents (water, ethanol, and 50% ethanol:water) were applied. The total phenolic compound (TPC) content, the main phenolic compounds and the in vitro antioxidant activity of the extracts were evaluated. The results were compared with those obtained by conventional decoction with water. The highest concentrations of TPC and antioxidant activity were obtained with 50% ethanol:water, followed by water. The extract obtained with 50% ethanol:water at 150 °C had a TPC concentration of 343.80 mg/g and presented the largest antioxidant activity (1488 and 4979 µmol Trolox/g extract, determined by DDPH and ABTS assay, respectively). These values were considerably higher than those obtained by conventional decoction. Ellagic acid, and ellagic and methyl ellagic acid glycosides were the main phenolic compounds found in the extracts. Therefore, was PLE demonstrated to be a selective and efficient technique to obtain extracts with high concentrations of phenolic compounds and high antioxidant activity form borututu roots.

Los insectos comestibles destacan como una fuente alternativa de proteínas. No obstante, durante el procesado de algunas especies se genera igualmente una fracción grasa relevante, como es el caso de las larvas de Hermetia illucens (mosca soldado negra), siendo un coproducto de gran interés para alimentación animal y humana. Por tanto, estudiar la influencia del procesado tecnológico sobre esta fracción grasa es clave para garantizar su calidad y uso en diversos fines. En el presente estudio, se llevó a cabo la evaluación del efecto de distintas formas de procesado de las larvas de H. illucens sobre la composición, calidad y bioactividad de la grasa obtenida. En concreto, se evaluaron dos formas de sacrificio (congelación o escaldado), seguido de dos formas de secado (horno o liofilización) y, de dos formas de desgrasado (prensado o extracción con fluidos supercríticos, SFE). Atendiendo al rendimiento de grasa, para todas las combinaciones de tratamientos, fue preferible el procedimiento de SFE, consiguiéndose el menor rendimiento al combinar escaldado-liofilización-prensa. De manera general, el perfil de ácidos grasos no sufrió cambios relevantes en función del procesado. No obstante, sí se evidenció una evidente lipólisis, con la consecuente generación de acidez libre, en función del tratamiento. Así, la aplicación de métodos no térmicos de congelación junto con liofilización dio lugar a la mayor degradación de la grasa en forma de ácidos grasos libres. Sin embargo, esta acidez fue minoritaria al aplicar métodos térmicos, como sacrificio por escaldado y secado por horno. Por otro lado, la combinación de congelación-liofilización-prensado produjo el mejor estado oxidativo inicial. De igual manera, el proceso de liofilización-prensado produjo las grasas más estables durante su almacenamiento a lo largo de 6 meses. En cuanto a la bioactividad de la grasa, se encontró una relación significativa entre la estabilidad oxidativa y la actividad antioxidante de la grasa, su

The interrelated effect of different slaughtering, drying and defatting methods of black soldier fly larvae (BSFL) on the lipid composition and properties of the fat was studied. Blanching and freezing were compared as slaughtering methods, oven or freeze-drying as drying methods, and mechanical pressing or supercritical fluid extraction (SFE) as defatting methods. The different modes of slaughtering, drying, and defatting, along with both binary and ternary interactions caused significant effects on processes yields, lipid composition, moisture content and thermal properties. Thus, considering the defatting degree and the yield in total valued products (defatted meal plus fat), the combination of blanching, freeze-drying plus mechanical pressing was the worst option (51.2% and 87.5%, respectively). In contrast, the other combinations demonstrated better and comparable efficiency, although SFE is preferable for defatting (83.2% and 96.9%, respectively). The content of major fatty acids (lauric, palmitic and myristic acids) was significantly affected by the BSFL treatments, although with unsignificant impact on the total saturated fatty acids content. To preserve the integrity of the fat, the combination of blanching and oven-drying was preferred, as non-thermal methods of slaughtering and drying caused intense lipolysis, releasing free fatty acids (FFA) in the range of 18.6-23.5%. To achieve the lowest moisture content in the fats (≤0.1%), oven-drying with mechanical pressing were desired, regardless of the slaughtering method; while values > 1% were reached for freezing, freeze-drying and SFE. Both differences in FFA and moisture contents caused different thermal behaviors in the samples. Specially, the melting temperature was lower for samples with higher FFA and moisture contents, with a notable difference when freezing, freeze-drying and SFE were combined (14.5 °C vs 30.6 °C, as the mean value for the rest of samples). The different modes of processing did not a

The aim of this study was to evaluate the modulating effect of the processing of Hermetia illucens larvae on the antibacterial activity of the fat related to lipolysis to free fatty acids (FFAs), mainly as lauric acid. Blanching and freezing were compared for slaughtering, oven or freeze-drying for drying, and mechanical pressing or supercritical fluid for defatting. Freezing plus freeze-drying produced the highest FFAs content (21%), mainly as lauric acid (11%). In agreement, freezing plus freeze-drying was also the most effective method to obtain fats with higher antibacterial activity, regardless of the defatting method. The antibacterial activity was significantly more relevant for gram-positive bacteria, having a strain-dependent character. The most effective fat was bactericidal for Listeria monocytogenes and Bacillus subtilis and reduced Staphylococcus aureus growth. Therefore, it is possible to selectively modulate the antibacterial activity of the H. illucens fat due to lipolysis by the processing methods of larvae.

[Simple Summary] In addition to the potential of edible insects as an alternative source of proteins, the production of insect meals generates a significant amount of fat that is also of interest for food and feed. The consideration of the impact of processing factors on the quality of insect fats, such as oxidative stability, is indispensable to fully realize their value as a co-product. Therefore, this study aimed to understand how the different successive modes of processing of Hermetia illucens larvae (black soldier fly larvae, BSFL), such as slaughtering (by blanching or freezing), drying (by oven-drying or freeze-drying) and defatting (by mechanical pressing or supercritical CO2 SFE), as well as their interactions, affect the oxidation of the resulting fat and defatted meal. The preferred procedure would be a combination of freezing with freeze-drying and mechanical pressing, especially the combination of freeze-drying and mechanical pressing, for the best oxidative quality during storage. Such oxidative stability was strongly related to the antioxidant activity of the fats, which was conditioned by the processing methods used for BSFL. Therefore, this study provides guidance on selecting the best combinations of slaughtering, drying, and defatting processes for BSFL to obtain fats with the best oxidative quality., The interrelation effect of slaughtering, drying, and defatting methods of BSFL on the oxidative quality of the derived fat was studied. Blanching and freezing were compared as slaughtering methods, followed by oven or freeze-drying for drying and mechanical pressing or SFE for defatting. The oxidative state and stability of the extracted fat and defatted meals were monitored immediately after their production, using peroxide value (PV) and Rancimat test, and over 24 weeks of storage. Slaughtering and drying methods had an independent effect on PV, with freezing and freeze-drying being the best methods. Mechanical pressing and SFE were comparable and superior to conventional hexane defatting. Interactions were observed between slaughtering and defatting, drying and defatting, and between all three factors. Generally, freeze-drying combined with any of the slaughtering and defatting methods resulted in the lowest PVs, with mechanical pressing being preferred. Freeze-drying plus mechanical pressing also produced the most stable fats during storage according to the evolution of PV, while the combination of blanching and SFE produced the least stable. A significant correlation was found between the PV at 24 weeks and the antioxidant activity of the fats. Contrary to storage assays, in accelerated Rancimat assays, freeze-dried samples were the least stable, which was partially attributed to the significant correlation with the acid values of the samples. Defatted meals followed a similar pattern to the extracted fat, except for worse oxidation for SFE defatting. Therefore, the different processing methods of slaughtering, drying, and defatting of BSFL differently affect lipid oxidation, with interactions between such successive steps.

Reversible and sub-lethal stresses to the mitochondria elicit a program of compensatory responses that ultimately improve mitochondrial function, a conserved anti-aging mechanism termed mitohormesis. Here, we show that harmol, a member of the beta-carbolines family with anti-depressant properties, improves mitochondrial function and metabolic parameters, and extends healthspan. Treatment with harmol induces a transient mitochondrial depolarization, a strong mitophagy response, and the AMPK compensatory pathway both in cultured C2C12 myotubes and in male mouse liver, brown adipose tissue and muscle, even though harmol crosses poorly the blood-brain barrier. Mechanistically, simultaneous modulation of the targets of harmol monoamine-oxidase B and GABA-A receptor reproduces harmol-induced mitochondrial improvements. Diet-induced pre-diabetic male mice improve their glucose tolerance, liver steatosis and insulin sensitivity after treatment with harmol. Harmol or a combination of monoamine oxidase B and GABA-A receptor modulators extend the lifespan of hermaphrodite Caenorhabditis elegans or female Drosophila melanogaster. Finally, two-year-old male and female mice treated with harmol exhibit delayed frailty onset with improved glycemia, exercise performance and strength. Our results reveal that peripheral targeting of monoamine oxidase B and GABA-A receptor, common antidepressant targets, extends healthspan through mitohormesis.

Edible insects are gaining attention as a source of bioactive compounds of great interest for health. To produce bioactive extracts from insects with a high concentration of bioactive compounds, defatting seems to be relevant for those fat-rich forms of insects as larvae. Supercritical fluid extraction (SFE) might be useful for both the defatting and subsequent extraction of the defatted sample to obtain bioactive extracts from edible insects. Different samples of black soldier fly (Hermetia illucens) larvae were defatted using SFE as an alternative to conventional defatting methods, and the optimal defatted flour was sequentially extracted by SFE employing different levels of ethanol as co-solvent. The resulting extracts were characterised by GC-MS-FID and their bioactivity was then assayed in terms of their antioxidant and pancreatic lipase inhibitory activities. The maximum fat yield by SFE (around 43%) was achieved at 450 bar, 60 degrees C and 100 g/min of CO2 flow, not being significantly different from conventional defatting with hexane. Extraction yield of SFE extracts increased along with the proportion of co-solvent. The composition of the extracts widely varied depending on the insect sample and the amount of co-solvent used, free amino acids and fatty acids being the two major chemical families among the different extracts. In terms of their bioactivity, a lipase inhibitory activity closer to 50% was exhibited by most extracts, which was favoured by the amount of co-solvent in one of the insect samples. Regardless of the significant differences observed between the insect samples, the exhibited DPPH radical inhibitory activity was weak. It is concluded that SFE is an interesting alternative to the conventional defatting of H. illucens, as well as an integrative and sequential process that allows producing bioactive extracts from this insect, especially towards the inhibition of the pancreatic lipase.

The mealworm (Tenebrio molitor Linnaeus 1758) is gaining importance as one of the most popular edible insects. Studies focusing on its bioactivities are increasing, although alternative forms of consumption other than the whole insect or flour, such as bioactive non-protein extracts, remain underexplored. Furthermore, the incidence of metabolic syndrome-related pathologies keeps increasing, hence the importance of seeking novel natural sources for reducing the impact of certain risk factors. The aim was to study the potential of a non-protein mealworm extract on metabolic syndrome-related pathologies, obtained with ethanol:water (1:1, v/v) by ultrasound-assisted extraction. We characterized the extract by gas-chromatography mass-spectrometry and assessed its hypolipidemic potential, its ability to scavenger free radicals, to attenuate the inflammatory response in microglial cells, to affect mitochondrial respiration and to enhance insulin sensitivity in mouse hepatocytes. The extract contained fatty acids, monoglycerides, amino acids, certain acids and sugars. The mealworm extract caused a 30% pancreatic lipase inhibition, 80% DPPH· scavenging activity and 55.9% reduction in the bioaccessibility of cholesterol (p = 0.009). The extract was effective in decreasing iNOS levels, increasing basal, maximal and ATP coupled respiration as well as enhancing insulin-mediated AKT phosphorylation at low insulin concentrations (p < 0.05). The potential of a non-protein bioactive mealworm extract against metabolic syndrome-related pathologies is shown, although further studies are needed to elucidate the mechanisms and relationship with compounds., [Simple Summary] Among edible insects, mealworm has gained attention due to its proven beneficial effects; however, mealworm products other than flour, such as concentrated forms or extracts different to proteins, are still rare. This novel food source can have a positive impact on multiple diseases such as via the metabolic syndrome. Our objective was to describe the composition of a mealworm extract and study its effect on metabolic syndrome-related pathologies. The extract interfered with the absorption of cholesterol and partially blocked the digestion of fats. It also reduced the inflammatory response, enhanced the sensitivity to insulin and showed strong antioxidant potential. Mealworm extract might be an interesting candidate for ameliorating the risk factors associated with the metabolic syndrome.

Nowadays, there is great interest in the discovery of food compounds that might inhibit gut microbial TMA production from its methylamine precursors. In this work, an innovative novel screening strategy capable of rapidly determining the differences in the metabolic response of Klebsiella pneumoniae, a bacteria producing TMA under aerobic conditions, to a library of extracts obtained from food and natural sources was developed. The proposed high-throughput screening (HTS) method combines resazurin reduction assay in 384-well plates and Gaussian Processes as a machine learning tool for data processing, allowing for a fast, cheap and highly standardized evaluation of any interfering effect of a given compound or extract on the microbial metabolism sustained by L-carnitine utilization. As a proof-of-concept of this strategy, a pilot screening of 39 extracts and 6 pure compounds was performed to search for potential candidates that could inhibit in vitro TMA formation from L-carnitine. Among all the extracts tested, three of them were selected as candidates to interfere with TMA formation. Subsequent in vitro assays confirmed the potential of oregano and red thyme hexane extracts (at 1 mg mL−1) to inhibit TMA formation in bacterial lysates. In such in vitro assay, the red thyme extract exerted comparable effects on TMA reduction (∼40%) as 7.5 mM meldonium (∼50% TMA decrease), a reported L-carnitine analogue. Our results show that metabolic activity could be used as a proxy of the capacity to produce TMA under controlled culture conditions using L-carnitine to sustain metabolism.