Until there is an effective implementation of COVID-19 vaccination program, a robust testing strategy, along with prevention measures, will continue to be the most viable way to control disease ...spread. Such a strategy should rely on disparate diagnostic tests to prevent a slowdown in testing due to lack of materials and reagents imposed by supply chain problems, which happened at the beginning of the pandemic. In this study, we have established a single-tube test based on RT-LAMP that enables the visual detection of less than 100 viral genome copies of SARS-CoV-2 within 30 min. We benchmarked the assay against the gold standard test for COVID-19 diagnosis, RT-PCR, using 177 nasopharyngeal RNA samples. For viral loads above 100 copies, the RT-LAMP assay had a sensitivity of 100% and a specificity of 96.1%. Additionally, we set up a RNA extraction-free RT-LAMP test capable of detecting SARS-CoV-2 directly from saliva samples, albeit with lower sensitivity. The saliva was self-collected and the collection tube remained closed until inactivation, thereby ensuring the protection of the testing personnel. As expected, RNA extraction from saliva samples increased the sensitivity of the test. To lower the costs associated with RNA extraction, we performed this step using an alternative protocol that uses plasmid DNA extraction columns. We also produced the enzymes needed for the assay and established an in-house-made RT-LAMP test independent of specific distribution channels. Finally, we developed a new colorimetric method that allowed the detection of LAMP products by the visualization of an evident color shift, regardless of the reaction pH.
Aiming at the enhancement of food products' nutritional and health value, the incorporation of nutraceuticals has attracted increasing interest in the last years. However, they often exhibit low ...water solubility and stability, limiting their direct incorporation into food products. Also, they show very low bioavailability due to limited bioaccessibility, poor absorption and/or chemical transformation within the gastrointestinal tract. This renders their health benefits extremely difficult to be realized by the consumers.
In the present review the recent innovations regarding the formulation and design of bio-based micro and nano-delivery systems to encapsulate nutraceuticals is discussed; it also gives an overview of the challenges associated to their development; and highlights some strategies to enhance nutraceuticals' bioavailability. An insight about delivery systems' potential toxicity (in particular at nano-scale) is also provided.
Recent developments in the design of bio-based delivery systems offer the possibility of stabilizing and enhancing nutraceuticals' functionality within food products. In fact, different strategies can be used to enhance nutraceuticals' bioavailability: i) nano-delivery systems, besides showing a huge potential for the protection of valuable nutraceuticals during food processing/digestion, can be used to increase their bioavailability; ii) absorption enhancement technologies have been successfully used to increase nutraceuticals' membrane permeation; and iii) excipient foods have been shown to improve nutraceuticals' biological activity. However, the application of these enabling technologies to food is hindered by very pertinent issues that can be summarized in the effective preservation/maximization of the nutraceuticals' bioactivity and safety, once inside the human body.
•Delivery systems allow nutraceuticals' protection and their incorporation in foods.•Bio-based delivery systems can be specifically designed to enhance nutraceuticals' bioavailability.•Bioavailability enhancement strategies include nano-delivery systems, absorption enhancers and excipient foods.•It is crucial to evaluate the safety of nano-delivery systems in food.•Development of delivery systems that provide nutraceuticals' greatest functionality and safety remains a challenge.
Colorectal cancer (CRC) recurrence is often attributable to circulating tumor cells and/or cancer stem cells (CSCs) that resist to conventional therapies and foster tumor progression. Isothiocyanates ...(ITCs) derived from
vegetables have demonstrated anticancer effects in CRC, however little is known about their effect in CSCs and tumor initiation properties. Here we examined the effect of ITCs-enriched
extracts derived from watercress and broccoli in cell proliferation, CSC phenotype and metastasis using a previously developed three-dimensional HT29 cell model with CSC-like traits. Both extracts were phytochemically characterized and their antiproliferative effect in HT29 monolayers was explored. Next, we performed cell proliferation assays and flow cytometry analysis in HT29 spheroids treated with watercress and broccoli extracts and respective main ITCs, phenethyl isothiocyanate (PEITC) and sulforaphane (SFN). Soft agar assays and relative quantitative expression analysis of stemness markers and Wnt/β-catenin signaling players were performed to evaluate the effect of these phytochemicals in stemness and metastasis. Our results showed that both
extracts and ITCs exert antiproliferative effects in HT29 spheroids, arresting cell cycle at G₂/M, possibly due to ITC-induced DNA damage. Colony formation and expression of LGR5 and CD133 cancer stemness markers were significantly reduced. Only watercress extract and PEITC decreased ALDH1 activity in a dose-dependent manner, as well as β-catenin expression. Our research provides new insights on CRC therapy using ITC-enriched
extracts, specially watercress extract, to target CSCs and circulating tumor cells by impairing cell proliferation, ALDH1-mediated chemo-resistance, anoikis evasion, self-renewal and metastatic potential.
Rosmarinic acid is a polyphenolic compound and main constituent of Rosmarinus officinalis and has been shown to possess antioxidant and anti‐inflammatory properties. We aimed to evaluate the ...anti‐inflammatory properties of rosmarinic acid and of an extract of R. officinalis in local inflammation (carrageenin‐induced paw oedema model in the rat), and further evaluate the protective effect of rosmarinic acid in rat models of systemic inflammation: liver ischaemia–reperfusion (I/R) and thermal injury models. In the local inflammation model, rosmarinic acid was administered at 10, 25 and 50 mg/kg (p.o.), and the extract was administered at 10 and 25 mg/kg (equivalent doses to rosmarinic acid groups) to male Wistar rats. Administration of rosmarinic acid and extract at the dose of 25 mg/kg reduced paw oedema at 6 hr by over 60%, exhibiting a dose–response effect, suggesting that rosmarinic was the main contributor to the anti‐inflammatory effect. In the liver I/R model, rosmarinic acid was administered at 25 mg/kg (i.v.) 30 min. prior to the induction of ischaemia and led to the significant reduction in the serum concentration of transaminases (AST and ALT) and LDH. In the thermal injury model, rosmarinic acid was administered at 25 mg/kg (i.v.) 5 min. prior to the induction of injury and significantly reduced multi‐organ dysfunction markers (liver, kidney, lung) by modulating NF‐κB and metalloproteinase‐9. For the first time, the anti‐inflammatory potential of rosmarinic acid has been identified, as it causes a substantial reduction in inflammation, and we speculate that it might be useful in the pharmacological modulation of injuries associated to inflammation.
In recent years many studies on cherries have revealed that they are rich sources of bioactive compounds with potential human health benefits. In this work, we evaluated the antioxidant activity and ...antiproliferative effect in human cancer cells of nine sweet cherries, including two traditional cultivars from Portugal (Saco and Morangão). Results obtained in biological assays, together with the phenolic composition of cherries, were submitted to principal component analysis (PCA) which allowed samples to be grouped in terms of their bioactivity. Saco cherry and two exotic cultivars (Ulster and Lapin) proved to have higher contents of phenolic compounds, highest antioxidant activity and were the most effective in inhibiting human cancer cells derived from colon (HT29) and stomach (MKN45). Correlation of the data obtained showed that anthocyanins were the major contributors to the antioxidant capacity and antiproliferative effect of cherries. Additionally, hydroxycinnamic acids (neochlorogenic acid, chlorogenic acid and
p-coumaroylquinic acid), flavan-3-ols (catechin and epicatechin) and flavonols (rutin and quercetin-3-glucoside) also play important roles in protection against oxidative stress.
•The most dominant polyphenol in blackthorn is neochlorogenic acid.•Blackthorn extracts inhibit α-amylase and α-glucosidase activity.•Antiproliferative effect on HT29 varies among blackthorn ...samples.•Quercetin glycosides and anthocyanins are the major contributors of the bioactive response.•PCA analysis is a useful tool to distinguish blackthorn genotypes according to chemical analysis.
The aim of this work was to investigate and compare the phenolic profile of 15 wild growing blackthorn (Prunus spinosa L.) genotypes from the slopes of Fruška Gora mountain in north Serbia. Their effect in inhibiting i) α-amylase and α-glucosidase activities and ii) colorectal cancer cell line (HT29) growth was also studied. Blackthorn fruit extracts exhibited high phenolic content being enrich in anthocyanins. Principal component analysis was used to correlate the bioactive response with phenolic composition. It was found that derivatives quercetin and anthocyanin peonidin are the major contributors of the inhibition of carbohydrates hydrolyzing enzymes as well as with the antiproliferative effect of blackthorn. Among all samples, the genotype from Beška locality showed the higher capacity in inhibiting alpha-amylase, alpha-glucosidase and HT29 cell growth. Because of high anthocyanin content and higher bioactive response, these genotypes could be recommended for the further cultivation and investigation.
During the last years, special interest has been focused on the recovery of bioactive compounds from food industry wastes in an effort to find added value applications for residues. In this work, ...Opuntia spp. residues from fruit juice production were explored as potential sources of natural chemotherapeutic ingredients towards colon cancer. Hydroalcoholic extraction and adsorption separation processes (with Amberlite XAD16 resin) were used to produce natural extracts from residues of Opuntia ficus-indica and Opuntia robusta and their antiproliferative effect was evaluated in human colon carcinoma HT29 cell line. Our results showed that the natural extracts efficiently inhibited cancer cell growth and induced cell cycle arrest in different checkpoints—G1, G2/M and S. The phytochemical constituents presented in the samples, namely betacyanins, flavonoids (isorhamnetin derivatives) and phenolic acids (ferulic acid) were identified as the main responsible compounds for the cell cycle arrest. Moreover, the extracts that presented the lowest effective dose values in terms of polyphenols (ED50 value<0.5mgGAE/mL) showed to increase intracellular ROS accumulation in HT29 cells, suggesting that cancer cell death may be induced by the pro-oxidant effect of these compounds.
•Phytochemical rich-extracts were obtained from residues of Opuntia juice production.•Opuntia juices and extracts presented distinct phenolic content and antioxidant value.•The antiproliferative effect on HT29 cells varied among samples.•Antiproliferative effect of juices well correlated with TPC, ORAC and HORAC.•Betacyanins, isorhamnetins and ferulic acid contributed to extracts' anticancer effect.
The aim of this work was to develop an innovative supercritical fluid (SCF)-assisted molecular imprinting method to endow commercial soft contact lenses (SCLs) with the ability to load specific drugs ...and to control their release. This approach seeks to overcome the limitation of the common loading of preformed SCLs by immersion in concentrated drug solutions (only valid for highly water soluble drugs) and of the molecular imprinting methods that require choice of the drug before polymerization and thus to create drug-tailored networks. In particular, we focused on improving the flurbiprofen load/release capacity of daily wear Hilafilcon B commercial SCLs by the use of sequential SCF flurbiprofen impregnation and extraction steps. Supercritical carbon dioxide (scCO2) impregnation assays were performed at 12.0MPa and 40°C, while scCO2 extractions were performed at 20.0MPa and 40°C. Conventional flurbiprofen sorption and drug removal experiments in aqueous solutions were carried out for comparison purposes. SCF-processed SCLs showed a recognition ability and a higher affinity for flurbiprofen in aqueous solution than for the structurally related ibuprofen and dexamethasone, which suggests the creation of molecularly imprinted cavities driven by both physical (swelling/plasticization) and chemical (carbonyl groups in the network with the C–F group in the drug) interactions. Processing with scCO2 did not alter some of the critical functional properties of SCLs (glass transition temperature, transmittance, oxygen permeability, contact angle), enabled the control of drug loaded/released amounts (by the application of several consecutive processing cycles) and permitted the preparation of hydrophobic drug-based therapeutic SCLs in much shorter process times than those using conventional aqueous-based molecular imprinting methods.
•The extraction of polyphenol compounds by using various solvents.•The determination of polyphenols, flavonoids, and condensed tannins.•The determination of the antioxidant activity using different ...methods.•The determination of the antibacterial activity.
Matricaria Pubescens (Asteraceae) is a well known medicinal plant used in the south west of Algeria against several diseases. This study was devoted to the determination of polyphenols, flavonoids, and condensed tannins contents of M. pubescens after their extraction using various solvents with different polarities (distilled water, aqueous acetone (50%), aqueous methanol (50%), aqueous ethanol (50%), acetone (100%), methanol (100%), and ethanol (100%)), as well as the determination of the antioxidant and the antibacterial activities of all obtained extracts. The results obtained indicate that the aqueous methanol (50%) and the aqueous ethanol (50%) extracts presented the highest contents of polyphenols. The antioxidant activity was evaluated using ferric reducing power, DPPH radical scavenging activity, ABTS radical scavenging activity, oxygen radical absorbance capacity (ORAC) and hydroxyl radical scavenging capacity (HOSC). The results showed that all extracts of M. pubescens, prepared using different solvent, have diverse antioxidant capacities, however aqueous methanol (50%), aqueous ethanol (50%) and aqueous acetone (50%) exhibited the highest antioxidant capacity. Extracts of M. pubescens exhibited an inhibiting capacity against the tested strains except: Escherichia coli ATCC 25922 and Acinetobacter baumannii 897. The aqueous methanol (50%) and the aqueous ethanol (50%) extracts were most effective; the greatest zone of inhibition was obtained with the aqueous ethanol (50%) extract against Methicillin-resistant Staphylococcus aureuscStaphylococcus aureusc. The minimum inhibitory concentration (MIC) with the value of 0.78mg/mL obtained against A. baumannii 610 using the aqueous methanol (50%) extract indicate that this plant is a potential natural antimicrobial agent. These biological activities could support the traditional use of M. pubescens.
β-Carotene formulations are very attractive as natural colorants as they provide additional value to the product due to their antioxidant and pro-vitamin activities. Application of β-carotene as ...colorants in beverages requires an appropriate formulation in order to stabilize the particles of β-carotene in suspension and provide the desired colour. This work presents a study of the formulation of β-carotene by precipitation from a pressurized ethyl acetate-in-water emulsion using modified OSA-starch refined from waxy maize as carrier material. Formulations of β-carotene with a high encapsulation efficiency of β-carotene (over 65% in most cases and as high as 90% with specific conditions), high antioxidant activity and a micellar particle size in the range of 300–600
nm were obtained. The concentration of modified starch and the organic solvent/water flow ratio were the process parameters with most influence on product properties.
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