Different methods for extraction of sage (Salvia officinalis) have been investigated by this research group. Hydroethanolic solid-liquid extracts presented the highest inhibitory capacity against pathogens. Alheira is a non-ready-to-eat sausage produced in Northern Portugal, which has in the past shown moderate prevalence of S. aureus. Thus, the aim of this study was to evaluate the antimicrobial effect of sage extract against S. aureus in alheira sausages exerted during the stage of maturation.

Lyophilised extract of sage aerial parts was obtained using ethanol 80% and distilled water as solvents, and kept at 15ºC for one week until use. Alheira batter was produced and added with 0.0%, 0.5%, 1.0% or 1.5% of sage (w/w), and stuffed in natural casings to reach an individual weight of ~85 g. They were then individually inoculated with a S. aureus ATCC 6538 culture to reach ~5 log CFU/g, while a non-inoculated control was kept. Sausages were hung in a chamber at 10 ºC/85% RH for 10 days. Water activity (aw), pH and S. aureus counts were determined every two days.

During maturation of inoculated and non-inoculated samples, aw in time followed an exponential decay model, dropping from 0.9913 to 0.9780, while pH dropped from 5.95 to 5.50. The inoculated S. aureus population decreased according to a downward concave Weibull model in 1.00, 1.12 and 1.23 log CFU/g, when alheira was formulated with 0.5, 1.0 and 1.5% sage extract, respectively. Oppositely, in the control treatment (0%), S. aureus counts did not change significantly (p>0.05) throughout maturation.

This work demonstrated that sage extract has a beneficial effect in controlling S. aureus in alheira. Individual phenolic compounds quantified for this extract also support this result, which is very relevant to artisanal producers since this frequently-found pathogen is mostly introduced to the batter through inadequate manipulation when mixing and stuffing.

Sustainability arises as a pivotal concept in different industries. Chestnut industry is one of the primary motors of the Portuguese fruit economy [1]. During chestnut processing, large amounts of by-products are produced, mostly shells. The wealth of chestnut shells in bioactive compounds, including vitamin E, amino acids and polyphenols, endowed with interesting biological activities, underlines its potential valorization as a substrate of natural antioxidants with attractive applications in the cosmetic field [1-3]. The implementation of eco-innovative technologies, such as supercritical fluids extraction (SFE), to recover bioactives has arisen as a priority for the industrial valorization of agro-residues.

The purpose of this study was to recover antioxidants from chestnut shells by SFE, aiming their reuse as new cosmetic principles. The extraction was performed at 60 °C, 350 bar and 15% co-solvent (ethanol) for 90 minutes. The extract was characterized regarding antioxidant and antiradical activities, elastase and hyaluronidase inhibition and in-vitro safeness on dermal cells (HaCaT and HFF-1). The skin permeation of polyphenols was screened using Franz cells coupled to human skin.

The antioxidant/antiradical properties (ABTS: 124.8 mg AAE/g dw; DPPH: 53.0% inhibition; FRAP: IC₅₀ = 204.8 μg/mL), and inhibitory activities on hyaluronidase (IC₅₀=54.4 mg/mL) and elastase (33.6%) reinforce the potential use of chestnut shells extract as an effective source of anti-aging ingredients. The in-vitro assay proved that the extract up to 100 µg/mL did not lead to a significant decrease of HaCaT and HFF-1 viability. Regarding ex-vivo assay, two polyphenols from SFE extract – ellagic acid (779.6 µg/g) and epicatechin (35.3 µg/g) – permeated the human skin after 8 h.

In conclusion, this study emphasizes the skin health effects and safety of C. sativa shells extract obtained by a green technology and proposes its incorporation in cosmetic formulations as an anti-aging ingredient.

Acknowledgements: This research was funded by FCT/MCTES through national funds (UIDB/50006/2020) and project PTDC/ASP-AGR/29277/2017 - Castanea sativa shells as a new source of active ingredients for Functional Food and Cosmetic applications: a sustainable approach, financially supported by national funds by FCT/MCTES and co-supported by Fundo Europeu de Desenvolvimento Regional (FEDER) throughout COMPETE 2020 - Programa Operacional Competitividade e Internacionalização (POCI-01-0145-FEDER-029277). Diana Pinto is thankful for her PhD grant (SFRH/BD/144534/2019) financed by POPH-QREN and subsidized by the European Science Foundation and Ministério da Ciência, Tecnologia e Ensino Superior. Francisca Rodrigues is thankful for her contract (CEECIND/01886/2020) financed by FCT/MCTES—CEEC Individual 2020 Program Contract.

References:

1. Pinto, D., et al., Castanea sativa shells: A review on phytochemical composition, bioactivity and waste management approaches for industrial valorization. Food Research International, 2021. 144: p. 110364.

2. Pinto, D., et al., Valorisation of underexploited Castanea sativa shells bioactive compounds recovered by supercritical fluid extraction with CO2: A response surface methodology approach. Journal of CO2 Utilization, 2020. 40: p. 101194.

3. Pinto, D., et al., Optimizing the extraction of phenolic antioxidants from chestnut shells by subcritical water extraction using response surface methodology. Food Chemistry, 2021. 334: p. 127521.

Nutritionally low diets have led to the increase of chronic non-communicable diseases (NCDs), particularly obesity and type II diabetes. Corn (Zea mays L.) and common beans (Phaseolus vulgaris L.) mixture have been tested as proper ingredients to manufacture healthy food products. However, but most of their biological potential reducing the risk of NCDs is mainly concentrated on its dietary fiber (DF), particularly for its content of phenolic compounds and its derived antioxidant capacity. This research aimed to characterize DF-aqueous extracts from nixtamalized corn and cooked common bean flour blends, evaluating their inhibitory potential against lipid and glucose metabolism in vitro. Two nixtamalized white corn flours (commercial: A; Northwestern white population, NWP: B) and one cooked common bean flour (Bayo Madero cultivar, C) were assessed on different proportions (80:20 and 70:30 corn:bean). An aqueous fiber extraction was conducted, and insoluble (IDF), soluble (SDF), and total dietary fiber (TDF) were quantified. IDF exhibited the highest total phenolics (30-45 µg eq. gallic acid/g sample), whereas SDF from 8020AC/8020BC displayed the highest content of condensed tannins. Chlorogenic and ellagic acids and (+)-catechin were the main identified phenolics by HPLC-DAD in the fiber extracts. SDF showed the highest antioxidant capacity levels (ABTS/DPPH), α-amylase, and pancreatic lipase inhibitions (20-25 %) among the fiber types. IDF exhibited the highest α-glucosidase inhibitions (up to ~75 %). In silico simulations indicated that epigallocatechin gallate had the highest binding affinity against the three assayed enzymes (-6.10 to -5.70 kcal/mol). Results suggested that DF extracts from nixtamalized NWP corn and cooked Bayo Madero common bean flours could be used as functional ingredients but incorporating these flours in suitable food products is required to validate their technological potential.

Millet is recognized as one of the beneficial cereals that can provide adequate nutrients and several studies have demonstrated that supplementation of millet has potential to possess valuable effects on the lifestyle disorder. However, the possible benefits of barnyard millet to the metabolic disorder disease are very limited . New insight in this area is important to improve the variety of healthy diet to the consumer especially to individual with metabolic syndrome. Thus, this study aimed to determine the effect of roasted whole grain barnyard millet flour on the diet-induced metabolic syndrome rats. We investigated the reversal sign of the metabolic syndrome of male Wistar rats on 10% addition of roasted whole grain barnyard millet flour to modified high carbohydrate high fats diet and corn starch diet for the final eight weeks of a 16-week protocol. The rats were divided into 5 group (n=8) and each of the group received special diet for 16 weeks: normal rats’ pallet (N), corn starch diet with barnyard millet (CM), corn starch diet (CS), high-carbohydrate high-fats diet with barnyard millet (HM) and high-carbohydrate high-fats diet (HF). After 16 weeks of diet regime, the intervention rats’ group CM and HM has a significant reduction (P<0.05) in fat mass, blood glucose tolerance between week 8 and week 16, and shows a significant difference (P<0.05) in weight, abdominal circumference, blood pressure between group CM with CS and group HM with HF. The results demonstrated the important potential of roasted whole grain barnyard millet health-promoting effect, specifically in reversing metabolic syndrome disease.

In this study, the effect of pH on gel properties of large yellow croaker (Pseudosciaena crocea) roe protein isolates (pcRPIs) was evaluated. The rheological properties, moisture-distribution and microstructure of pcRPIs gels were also analyzed. The results showed that pcRPIs failed to form gels at pH 4-6, while indicated gelation profile at pH 7-9. The optimum pH for pcRPIs to form gels was pH 8 with the gel point at 84 ℃, and the G’ of the pcRPIs gel formed at pH 8 reached 535.4 Pa at the concentration of 100 mg/mL. In addition, as pH increased, pcRPIs gels had smoother surface and more continuous network structure, and the water holding ability of pcRPIs gels increased. Moreover, hydrophobic interactions were the main forces in the formation of pcRPIs gels. These results indicated that pcRPIs had potential to be used as functional materials in the food industry, especially some gel products.

Manufacturers have been deeply involved in the development of dried fruits in last decade. Therefore, there is an increase in the range of dried fruit on the market right now. It is essential to highlight that there is not daily recommendation intake of dried fruit. The aim of the present study was to compare the labeling information, physico-chemical parameters, antioxidant capacity and sensory profile among: i) different dried fruits (apple, mango, pineapple, tomato, fig, coconut, banana and red cranberry) and, ii) different trademarks of each type of dried fruits (n=3). Depending on the type of fruit, the labelling information focused on “clean label trend” varies. The ingredient list depends on physico-chemical parameters such as water activity, humidity and sugar content. Among brands, significant differences (p-value < 0,05) of color coordinates, antioxidant capacity, organic acid profile, sugar profile, and sensory attributes (texture, fruit ID, basic tastes) were found. As to nutritional quality, it is essential to highlight a high content of sugars was found in all samples (>30 g/100 g). On the other hand, a high content of fiber was found (> 10 g/100g) in dried coconut, apple and tomato samples.

Spent coffee grounds (SCGs) are a by-product of the food beverage industries, which contains a rich source of polysaccharides. This research aims to study the extraction of polysaccharides from SCGs by environmentally friendly technique of pressurized hot water. The process optimization was investigated by response surface methodology (RSM) to obtain the highest extraction yield. The investigated parameters were temperature of 80-120 °C, pressure of 4-12 bars, and extracted time of 60-180 min. The pressurized hot water showed an efficient technique to recover polysaccharides from SCGs. The best condition to provide the highest yield of 13.71±53% was the temperature of 120 °C, the pressure of 4 bar, and the extracted time of 60 min. In addition, the total phenolic content from obtained polysaccharide was 11.13 ±1.33 mg gallic acid equivalent (GAE)/g dry SCG.

Grey oyster mushroom (Pleurotus sajor-caju (Fr.) Singer) is a popular edible mushroom in Thailand due to its high nutritional and medicinal benefits. This study aims to study the effects of temperature (100-140 °C), pressure (4-7 bar), and extraction time (20-60 min) on the extraction crude polysaccharides with environmentally friendly pressurized hot water. The extraction condition was optimized by the maximize yield using response surface method based on a central composite design (CCD). The temperature was the main effect on the increase of extracted yield. The optimum extraction condition was 140 °C, 10 bar, and 26.79 min with a corresponding yield of 31.31±2.55%. Under these conditions, the total phenolic content of crude polysaccharides was 401±8.24 mg GAE/g dry mushroom. In addition, the total glucan content was indicated as 34.50 ±1.79 g/100 g dry mushroom, which separated as 32.47±1.95 mg/100 g of β-glucans and 2.04±0.98 mg/100 g of a-glucans.

Alternative cancer treatment with dietary supplements is attractive worldwide, and mushrooms, as abundant source of bioactive compounds, have confirmed therapeutic effects. Moreover, cancer fungotherapy, as a promising scientific field, focuses on investigation of fungal products with prominent anticancer potential and target-specific activity. Beside their cytotoxicity, effects of mushrooms regarding regulation of crucial steps in cancer metastasis are poorly investigated. Our study aimed to analyze effects of two edible and medicinal mushroom species on protein markers of migration and invasion . Methanol extracts of commercially cultivated edible and medicinal mushroom species Phellinus linteus (PL) and Lentinus edodes (LE) were examined in two selected concentrations (10 and 50 µg/mL) for their antimigratory/invasive potential on two colorectal carcinoma cell lines (HCT-116, SW-480) and effects were tracked after 24 h. Expression and localization of pro/antimigratory proteins E-cadherin, N-cadherin and Vimentin were done by using cytoimmunochemistry, and concentration of matrixmetalloproteinase 9 (MMP-9) was determined using colorimetric ELISA assay. According to the results, PL and LE induced obvious cell selective effects, whereat LE had stronger effect on HCT-116 cells, whilst PL exerted more prominent effect on SW-480 cells. LE increased expression of antimigratory marker E-cadherin and significantly decreased level of promigratory/proinvasive proteins N-cadherin and Vimentin in HCT-116 cells. Meanwhile, PL induced similar response in SW-480 cells, increasing E-cadherin and lowering N-cadherin and Vimentin. However, both applied treatments significantly decreased concentration of proinvasive protein MMP-9 in both tested cell lines. Once again, cell selectivity was observed regarding effects of these treatments. Namely, the best effect on SW-480 cell line exerted PL which significantly lowered level of MMP-9, and LE was more potent in reducing the level of this proinvasive protein in HCT-116 cells. Further studies should to be conducted regarding these mushrooms that possess obvious and important antimigratory/antiinvasive potential, esspecially their application as food supplements.

Kelulut (stingless bee) honey (KH) possesses a wide range of benefits for human consumption, such as high nutritional value, antimicrobial potency, high antioxidant value, and can exhibit medical effects. Due to its high value, this premium honey is often adulterated with different types of cheaper sugars, causing low nutrients and potential food safety threats in the final product. This emerging problem has driven many types of research to detect adulteration in honey. However, currently, there is minimal research about sugar-based adulteration of KH in Malaysia. This study aims to determine the physicochemical and rheological properties of sugar-based adulterated KH from the species Heterotrigona Itama. Adulterated samples were prepared using pure honey mixed with different concentrations of high fructose corn syrup (HFCS), i.e., 10%, 20%, 30%, 40% and 50%. Water activity, colour, total soluble solids, pH, turbidity and viscosity of KH were determined according to Official Methods of Analysis (AOAC), Malaysia Standard, MS 2683: 2017: KH – specification (Standard 2017) and harmonized methods of the International Honey Commission. In addition, the primary sugar composition (fructose, glucose and trehalulose) was determined by high-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD). This study shows that the increasing percentage of HFCS addition in the KH samples significantly increase (P <0.05) the total soluble solids, colour, pH and turbidity; meanwhile, the water activity is reduced significantly (P <0.05). The viscosity of honey increased gradually with HFCS and glucose and fructose content while trehalulose content decreased. This research found that the physicochemical properties of sugar-based adulterated and authentic KH can be differentiated. This data is vital for the governing bodies to ensure that KH sold in the markets are free from sugar-based adulterants.