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The main purpose of the present study was to determine the effect of associating an optimized ultrasound-assisted extraction (UAE) protocol with enzyme-assisted extraction (EAE) in aqueous media, using the dried berries of Hippophae rhamnoides L. (sea buckthorn) as plant material. A specialized software was used for the determination of potential optimal extraction parameters, leading to the development of four optimized extracts with different characteristics (UAE ± EAE). For these extracts, buffered or non-buffered solutions have been used, with the aim to determine the influence of adjustable pH on extractability. As enzymatic solution, a pectinase, cellulase, and hemicellulase mix (2:1:1) has been applied, acting as pre-treatment for the optimized protocol. The highest extractive yields have been identified for non-buffered extracts, and the E-UAE combination obtained extracts with the highest overall in vitro antioxidant activity. The HPLC-MSn analysis demonstrated a rich composition in different types of isorhamnetin-O-glycosides, as well as some quercetin-O-glycosides, showing a high recovery of specific flavonol-type polyphenolic species. Moreover, we have tentatively identified two flavanols (i.e., catechin and epigallocatechin) and one flavone derivative (i.e., luteolin).

Polyphenols were extracted from Thuja occidentalis L. bark and characterized in terms of phytochemical composition and biological activity. The optimization process investigated the impacts of extraction duration, ultrasound amplitude, and ethanol concentration on the total phenolic content in the extract. The antioxidant capacity was examined using DPPH and ABTS assays, and anti-enzyme activity against alpha-glucosidase, alpha-amylase, lipase, acetylcholinesterase, and tyrosinase was determined. The experimental model revealed optimal extraction parameters: a hydroethanolic solvent with 44% v:v ethanol, a 15-minute extraction time, and a 40% ultrasound amplitude. These parameters were validated and confirmed by reaching the target Total Phenolic Content (TPC) predicted by MODDE software. The resulting lyophilized extract exhibited a high polyphenolic content (161.84 ± 23.48 mg GAE/g dry extract) and demonstrated strong antioxidant properties. Notably, it showed significant inhibition of alpha-glucosidase and alpha-amylase, suggesting potential antidiabetic effects, though its inhibition of tyrosinase activity was relatively weak. These findings provide a foundation for further investigations to confirm the qualitative and quantitative presence of specific polyphenolic metabolites in the extract and elucidate the mechanisms underlying its bioactive properties.

Literature is limited regarding the potential use of the bark of Maclura pomifera as a source of bioactive compounds. The influence of different extraction methods was assessed for the chemical and biological properties of extracts obtained from the bark and periderm of M. pomifera. The extraction process was completed using three methods: microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), and sonicator-assisted extraction (SAE). The extracts were characterized by total polyphenol content. The antioxidant capacity was evaluated using DPPH and ABTS methods, the antimicrobial effect was tested against Gram-positive and Gram-negative bacterial strains, and the inhibition of enzyme activity was conducted using α-glucosidase, lipase, and α-amylase assays. The extracts obtained from the periderm exhibited a lower total polyphenolic content compared to those obtained from the bark. The highest antioxidant activity using the DPPH method was observed in the MAE bark extracts and when using the ABTS method, and the highest activity was observed in the UAE bark extracts. Enzymatic activity was most strongly inhibited by the SAE bark extract. The most pronounced antibacterial potential was observed in the MAE and UAE bark extracts, with minimum inhibitory concentration values obtained below 1 mg/mL.

Non-communicable diseases (NCDs) cause 41 million deaths annually, accounting for 74% of global fatalities. The so-called Mediterranean diet, with its especially significant consumption of olive oil, has shown promising results in reducing the risk of developing NCDs, such as cardiovascular, liver, or bone diseases. In the context of the nutritional health benefits of foods, phenolic compounds such as olive oil’s main components, oleuropein (OLE) and hydroxytyrosol (HT), have been shown to possess different beneficial effects. However, no systematic review has evaluated the health-promoting effects of OLE and HT until now. Consequently, this systematic review analyzed 12 human randomized controlled trials (RCTs), involving 683 participants, to assess the effects of supplements, pure compounds, or enriched foods containing OLE and HT regarding systemic health outcomes, including CVD risk factors, liver parameters, and bone, joint, and cognitive health. The review found contrasting but encouraging results, with some studies reporting significant modulation of body weight, lipid profile, and glucose metabolism, and improvements in bone, joint, and cognitive functions. The studies described different dosages and forms of supplementation, ranging from 5 mg/d HT to 990 mL/d olive leaf infusion (320.8 mg OLE and 11.9 mg HT), highlighting the need for further research to determine the optimal dosing and duration. Despite the mixed outcomes, OLE and HT supplementation show potential for improving some of the cardiometabolic health outcomes and bone, joint, and cognitive health. However, further studies are necessary to understand their benefits better and address existing limitations.

Nutraceuticals and functional foods are composed of especially complex matrices, with polyphenols, carotenoids, minerals, and vitamins, among others, being the main classes of phytochemicals involved in their bioactivities. Despite their wide use, further investigations are needed to certify the proper release of these phytochemicals into the gastrointestinal medium, where the bioaccessibility assay is one of the most frequently used method. The aim of this review was to gather and describe different methods that can be used to assess the bioaccessibility of nutraceuticals and functional foods, along with the most important factors that can impact this process. The link between simulated digestion testing of phytochemicals and their in vitro bioactivity is also discussed, with a special focus on the potential of developing nutraceuticals and functional foods from simple plant materials. The bioactive potential of certain classes of phytochemicals from nutraceuticals and functional foods is susceptible to different variations during the bioaccessibility assessment, with different factors contributing to this variability, namely the chemical composition and the nature of the matrix. Regardless of the high number of studies, the current methodology fails to assume correlations between bioaccessibility and bioactivity, and the findings of this review indicate a necessity for updated and standardized protocols.

Corn silk, derived from the stigmas of corn (Zea mays, Poaceae) flowers, possesses valuable bioactive compounds that can be harnessed for therapeutical, nutritional, and industrial purposes. This research aimed to optimize the extraction of bioactive compounds from corn silk, specifically flavonoids and total antioxidant capacity, by comparing it with classical extraction (infusion and maceration). The study identified significant factors influencing total flavonoid content (TFC) and antioxidant activity (assessed through an ABTS assay), including ethanol concentration, ultrasonic amplitude, and their quadratic interactions. This optimized combination of independent variables (37.89% ethanolic concentration, 48.58% ultrasound amplitude, and an extraction time of 38.72 min) yielded a balanced extraction, maximizing both TFC and antioxidant activity. Further analyses revealed significant total phenolic content, total flavonoid content (50.69 ± 0.36 mg GAE, 23.54 ± 0.01 mg QE, and 78.33 ± 0.94 mg TE/g extract, respectively), and potent in vitro antioxidant activity, affirming the efficacy of the optimized extraction. The study also explored the inhibitory effects on α-glucosidase (IC50 = 411.67 ± 25.39 µg/mL), acetylcholinesterase (IC50 = 1166.4 ± 399.38 µg/mL), and pancreatic lipase (IC50 = 1519.67 ± 209.04 µg/mL), showcasing potential antidiabetic and neuroprotective properties. While tyrosinase inhibition activity was not observed, the research emphasizes the nutraceutical potential of corn silk, offering valuable insights for its utilization and further exploration in the pharmaceutical and functional food industries.

Thymus comosus Heuff ex. Griseb. (Lamiaceae) is a wild thyme species endemic for Romanian Carpathian areas, frequently collected as substitute for collective herbal product Serpylli herba, cited as antibacterial and diuretic remedy in traditional medicine. The present study aimed to evaluate the in vivo diuretic effect and in vitro antimicrobial properties of three herbal preparations (infusion—TCI, tincture—TCT and an hydroethanolic extract prepared through an optimized ultrasound-assisted method—OpTC) obtained from the aerial parts of T. comosus Heuff ex. Griseb, also evaluating their comprehensive phenolic profile. In vivo diuretic effect was tested using Wistar rats treated orally with each herbal preparation (125 and 250 mg/kg dispersed in 25 ml/kg isotonic saline solution) and quantified based on cumulative urine output (ml), diuretic action and diuretic activity. Additionally, sodium and potassium excretion were monitored using a potentiometric method with selective electrodes. In vitro antibacterial and antifungal activities were assessed using p-iodonitrotetrazolium chloride assay against six bacterial strains and six fungal strains by monitoring minimum inhibitory concentration (MICs), minimum bactericidal concentrations (MBCs) and minimum fungicidal concentrations (MFCs). Finally, phenolic profile of the aforementioned herbal extracts was evaluated using an ultra-high-pressure liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) method to check the impact of the different preparations on the most abundant and significant compounds. All the extracts exerted a mild diuretic action, TCT and OpTC inducing the most intense diuretic effect. Both herbal preparations produced a statistically significant, dose-dependent and gradual increase of the urine output, the effect being more intense at 24 h (6.63–7.13 ml/24 h). Potentiometric evaluation of urine samples collected from treated rats revealed a clear and mild natriuretic and kaliuretic effect after the administration. In terms of antimicrobial activity, E. coli (MIC—0.38 mg/ml), B. cereus (MIC—0.75 mg/ml)), Penicillium funiculosum and P. verrucosum var. cyclopium (MIC—0.19 mg/ml) showed the greater sensitivity to the tested extracts, respectively. UHPLC-HRMS screening showed that the bioactive potential of T. comosus herbal preparations was likely related to the higher amounts of phenolic acids (including rosmarinic acid), flavonoids (mainly flavones and derivatives) and other phenolics (such as different isomers of salvianolic acids) in their composition. The obtained results support the ethnopharmacological evidence regarding the mild diuretic and antibacterial potentials of the endemic wild thyme T. comosus, this study being the first one that assessed the aforementioned bioactivities for this species.

Pinus cembra L., also known as Swiss pine, is one of the lesser studied coniferous species, despite literature pointing out its great potential to be used for medical purposes due to its high contents in active phytochemicals. The aim of this study is to phytochemically assess rhytidome and periderm extracts obtained from Swiss pine from various locations and altitudes, so as to be able to deduce the best locations for harvesting samples with the highest biological activity. After the plant was analyzed histo-anatomically, hydroalcoholic extracts were obtained using ultrasounds, a rotary evaporator, and dry freezing. After determining the total polyphenolic content (TPC) in each sample, they were tested for antioxidant and enzymatic activity, while taking note of the influence of the varying altitudes and different harvesting regions on the intensity of each activity. The results from the TPC analysis show that rhytidome samples collected from the highest altitude (2429 m) displayed the highest content of polyphenols, with a general tendency for the amount of polyphenolic content to be directly correlated to altitude, this finding being further supported by the antioxidant activity also growing directly proportionally with the altitude. Enzymatic inhibition activity was found to be moderate for α-glucosidase and low for α-amylase. Following our findings, we can safely say that Pinus cembra L. extracts can be a great source of compounds with antioxidant activity; however, further studies are warranted to best determine the true potential of this species.

The aim of this study was to reveal the antiproliferative effect of two tomato extracts on colorectal cancer cells in vitro, and to show their influence on the secretion of growth factors involved in tumor neoangiogenesis: VEGF and Endothelin-1, in order to reveal some of the inhibitory mechanisms exerted by the lycopene-containing tomato extracts. Human colon cancer cell lines DLD1 and HT29 were treated with two different tomato extracts, obtained from fresh and frozen tomato fruits. The cytotoxicity of the compounds was measured using the MTT test, while the expression of VEGF and Endothelin-1 was assessed by ELISA. Results showed that the extract from frozen tomatoes was more cytotoxic as the analogue from fresh tomatoes on both cell types, and both extracts were more toxic against DLD-1 cell line. VEGF and Endothelin-1 expression was also reduced by the tomato extracts, especially in DLD1. In vitro tests demonstrated that the lycopene-containing tomato extracts reduce the growth of malignant cells by tumor neovascularization hindrance, opening new perspectives to their employment in human therapeutic purposes.

Ethnopharmacological relevance:Salvia glutinosa, also known as the glutinous sage, has been used in Romanian folk medicine in the treatment of inflammation, injuries, and mild infections. However, there is no direct scientific evidence to demonstrate these activities.Aim of the Study: The present research was based on evaluating antioxidant, antiproliferative, and α-glucosidase inhibitory activity of S. glutinosa extracts, as well as the in vivo anti-inflammatory activity.Materials and Methods: Infusions and 70% (v:v) ethanol solution extracts of S. glutinosa stems and leaves, collected from two different locations in Romania, were prepared. Ten phenolic compounds were identified and quantified using the LC-DAD-ESI/MSn method, and total phenolic and flavonoid content, as well as in vitro antioxidant (DPPH, ABTS, and FRAP assays), antiproliferative, anti-inflammatory and alpha-glucosidase inhibitory activities were determined. A rat model of induced inflammation with turpentine oil was used for the examination of in vivo effects of the extracts, using diclofenac as an anti-inflammatory control.Results: The highest inhibitory α-glucosidase activity was determined to be IC50 = 0.546 mg/ml for the hydroalcoholic extract made with plant material collected on the road to Sighișoara. The highest cytotoxic activity against HepG2 cell line was determined to be GI50 = 131.68 ± 5.03 μg/ml, for the hydroalcoholic extract made with plant material from Sighișoara. In vivo administration of extract (200 mg lyophilized powder/ml) showed a significant reduction of NO production.Conclusion: Our findings indicate that S. glutinosa extracts exhibit antioxidant, α-glucosidase inhibitory activity, as well as a modest cytotoxic effect on HepG2 cell line. By in vivo administration, the extracts show anti-inflammatory and antioxidant activity, which correlates with the traditional use of the species. The environmental conditions seemed to induce important changes in the chemical composition and the bioactivity of the herbal preparations derived from S. glutinosa.

BackgroundShallot (Allium ascalonicum L.) is a traditional plant species used throughout the world both for culinary purposes and as a folk remedy. To date (i.e., April 2022), there is no report on the main pharmacological activities exerted by shallot preparations and/or extracts.Scope and ApproachThe aim of this study was to comprehensively review the pharmacological activities exerted by shallot, with rigorous inclusion and exclusion criteria based on the scientific rigor of studies. Prisma guidelines were followed to perform the literature search.Key Findings and ConclusionsThe literature search yielded 2,410 articles of which 116 passed the required rigorous criteria for inclusion in this review. The extracts exert a potent antioxidant activity both in vitro and in vivo, as well as a strong inhibitory capacity on various pathogens with relevant implications for public health. Moreover, shallot can be used as adjuvant therapy in cardiovascular diseases, diabetes, cancer prevention, and other non-communicable diseases associated with inflammatory and oxidative pathways. Future studies investigating the chemical composition of this species, as well as the molecular mechanisms involved in the empirically observed pharmacological actions are required.

Abstract Household responses to COVID-19 in different corners of the world represent the primary health care that communities have relied on for preventing and mitigating symptoms. During a very complex and confusing time, in which public health services in multiple countries have been completely overwhelmed, and in some cases even collapsed, these first-line household responses have been quintessential for building physical, mental, and social resilience, and for improving individual and community health. This editorial discusses the outcomes of a rapid-response preliminary survey during the first phase of the pandemic among social and community contacts in five metropolises heavily affected by the COVID-19 health crisis (Wuhan, Milan, Madrid, New York, and Rio de Janeiro), and in twelve rural areas or countries initially less affected by the pandemic (Appalachia, Jamaica, Bolivia, Romania, Belarus, Lithuania, Poland, Georgia, Turkey, Pakistan, Cambodia, and South Africa). We summarized our perspectives as 17 case studies, observing that people have relied primarily on teas and spices (“food-medicines”) and that there exist clear international plant favorites, popularized by various new media. Urban diasporas and rural households seem to have repurposed homemade plant-based remedies that they use in normal times for treating the flu and other respiratory symptoms or that they simply consider healthy foods. The most remarkable shift in many areas has been the increased consumption of ginger and garlic, followed by onion, turmeric, and lemon. Our preliminary inventory of food medicines serves as a baseline for future systematic ethnobotanical studies and aims to inspire in-depth research on how use patterns of plant-based foods and beverages, both “traditional” and “new”, are changing during and after the COVID-19 pandemic. Our reflections in this editorial call attention to the importance of ethnobiology, ethnomedicine, and ethnogastronomy research into domestic health care strategies for improving community health.

The members of Cystus genus are perenial shrubs with a well-established use in traditional medicine. Among these, C. creticus is the most popular, herbal preparations obtained from its aerial parts being recognized as antimicrobial, antitumor and anti-inflammatory agents. The present study aimed to evaluate phytochemical profile and bioactive potential of aqueous and hydroethanolic extracts of C. creticus aerial parts harvested from two different areas of Rhodes island. LC-DAD-ESI/MSn analysis revealed the presence of myricetin and quercetin glycosides as main compounds, especially in aqueous extracts, being probably responsible for their enhanced antioxidant and antimicrobial potential. On the other side, hydroethanolic preparations exerted a strong anti-inflammatory and anti-biofilm activity. Our findings suggest that the use of solvents with intermediate polarity can assure the best recovery of bioactive compounds from C. creticus, increasing the extraction yield for other non-phenolic compounds which can enhance therapeutic potential of the extract through a synergistic action.

The pomegranate is an emerging functional food which is nowadays becoming more and more commercially attractive. Each part of this fruit (peels, arils and seeds) has a specific phytocomplex, rich in anti-oxidant and anti-radical compounds. Among these, punicalagin and ellagic acid continue to be widely studied for their numerous beneficial effects on human health (anti-inflammatory effects, anti-diabetes activity, cardio-protection, cancer prevention). Despite their exceptionally valuable composition and high adaptability to different climatic conditions, pomegranate fruits are highly susceptible to splitting during different stages of ripening, so much so that an estimated 65% of the production may be lost. A “zero-kilometer” approach should therefore be adopted to utilize such a valuable product otherwise destined to be downgraded or even incinerated, with a very high environmental impact. The aim of this work is to highlight and compare the compositional differences between whole and split pomegranates belonging to the cultivar Dente di Cavallo, grown in Apulia (Italy), to assess a valuable role for this split fruit usually considered as waste. The arils and peels are subjected to extraction procedures and the extracts analyzed by CIEL*a*b*, HPLC-DAD and HS-SPME/GC-MS. Moreover, an assessment of the inhibitory activity against α-glucosidase, acetylcholinesterase and tyrosinase enzymes has also been applied. The data show a better chemical profile in split fruits (namely 60% more anthocyanin content than intact fruit) with very interesting results in terms of α-glucosidase inhibition. The juices obtained by squeezing are also compared to commercial juices (“Salus Melagrana” and “La Marianna”) processed from the same cultivar and subjected to the same protocol analysis.

Phytoremediation is an effective method used to control the accumulation of certain contaminants found in industrial or city wastewater. Among the species with high efficacy are Eichhornia crassipes (water hyacinth), Lemna minor (common duckweed), and Pistia stratiotes (water lettuce). In this study, the application of these species in the context of two municipal wastewater treatment facilities in Cluj County, Romania, is evaluated. To determine the efficacy of bioaccumulation, we measured the content of nitrogen species (ammoniacal nitrogen, nitrites, and nitrates), phosphorous, iron, and chromium before and after the addition of plant material to effluent and treated wastewater. The results showed that E. crassipes, L. minor, and P. stratiotes presented high phytoremediation yields for these common wastewater pollutants after one week of contact, with yields as high as 99–100% for ammoniacal nitrogen, 95% for phosphorous, 96% for iron, and 94% for chromium. However, the remediation capacity for nitrate and nitrite was less significant.

An optimized ultrasound-assisted extractive method was developed to obtain a polyphenol-enriched extract from the aerial parts of Thymus comosus Heuff. ex Griseb. et Schenk. Optimization process was conducted based on Design of Experiment (DoE) principles, determining the influence of three independent variables (time, ultrasound amplitude, ethanol concentration) on the total phenolic content of the extract (dependent variable). Additionally, the phenolic composition of the extract was characterized through UHPLC-HRMS, revealing beside the most abundant flavonoid-type compounds the presence of salvianolic acids C, D and L in high amounts. Phytochemical profile of the extract was correlated with its antioxidant activity (tested through five complementary assays) and enzyme-inhibitory potential, showing important antiglucosidase and anticholinesterase effects. Overall, it was concluded that the developed method is suitable for obtaining a good recovery of both phenolic and non-phenolic compounds from Thymus comosus aerial parts, and their presence in the optimized extract is responsible for its pharmacological potential.

Wild thyme aerial parts (Serpylli herba) are recognized as a valuable herbal product with antioxidant, anti-inflammatory, and antibacterial effects. Although pharmacopoeial regulations allow its collection exclusively from Thymus serpyllum, substitution with other species is frequent in current practice. This study analyzed the phenolic composition, antioxidant, and enzyme-inhibitory and antimicrobial activity of the hydroethanolic extracts obtained from five Romanian wild thyme species (Thymus alpestris, T. glabrescens, T. panonicus, T. pulcherimus and T. pulegioides). The analysis of individual phenolic constituents was performed through LC-ESI-DAD/MS2, while for the in vitro evaluation of antioxidant potential, TEAC, FRAP, DPPH, TBARS and OxHLIA assays were employed. The anti-enzymatic potential was tested in vitro against tyrosinase, α-glucosidase and acetylcholinesterase. High rosmarinic acid contents were quantified in all species (20.06 ± 0.32–80.49 ± 0.001 mg/g dry extract); phenolic acids derivatives (including salvianolic acids) were confirmed as the principal metabolites of T. alpestris and T. glabrescens, while eriodictyol-O-di-hexoside was found exclusively in T. alpestris. All species showed strong antioxidant potential and moderate anti-enzymatic effect against α-glucosidase and acetylcholinesterase, showing no anti-tyrosinase activity. This is the first detailed report on the chemical and biological profile of T. alpestris collected from Romanian spontaneous flora.

The bark resulted as a by-product after different Quercus sp. processing is a valuable alternative source of phenolic compounds (mainly tannins). Hence, the focus of the present work was to obtain eight extracts from the rhytidome of the less-studied Q. dalechampii and Q. frainetto and characterize them in terms of chemical and bioactive profiles. Ultrasound and microwave-assisted extraction methods were used for the preparation of the extracts. Total phenolic and tannin contents were assessed through classic spectrophotometric methods, while several individual phenolic compounds were identified and quantified using UPLC-PDA. Antioxidant, enzyme-inhibitory, antibacterial, and cytotoxic activities were tested using in vitro assays; additionally being evaluated was the ability of the extracts to inhibit the adherence of MRSA to suture wires. The UPLC analysis confirmed the presence of gallic acid, catechin, taxifolin, vanillic acid, epicatechin, and caffeic acid. The results showed that tested extracts were able to exert cytotoxic effects, at 6% and 3% concentrations, on confluent cells. The tested solutions inhibit α-glucosidase activity and the antibacterial potential suggested a mild to moderate effect against the Gram-positive strains. Overall, the obtained results revealed rich phenolic and tannin contents for the extracts obtained from both species through microwave-assisted extraction, probably responsible for their mild antibacterial and cytotoxic effects.

Processing wood leads to the accumulation of considerable amounts of forest waste, such as branches, leaves and especially bark. These byproducts can be exploited for their high content of biomolecules. Our aim was to histo-anatomically and phytochemically characterize the older oak bark and evaluate the potential biological activities. After the histo-anatomical characterization and processing the plant material, the aqueous and hydroalcoholic extracts were obtained using microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE). After the quantification of the total polyphenol content (TPC) and the total tannins content (TTC), the bark extracts were evaluated for antioxidant, antimicrobial and enzyme inhibitory activity. In the transverse section, the rhytidome (the succession of periderms) and the dead secondary phloem can be observed. The results of the polyphenol content of the oak bark extracts show that the hydroethanolic oak bark obtained using MAE has the highest absolute value of the TPC (347.74 mg/g). In comparison with the extracts obtained using UAE, the bark extracts obtained using MAE showed a significantly higher TPC. The greatest antioxidant and antibacterial activity were also recorded for the tested extracts. For gram-positive bacteria, the MBC values ranged from 0.3 (S. aureus) to 5 mg/mL and the oak bark extract obtained using MAE showed good activity against C. krusei (MIC—2.5 mg/mL). Among the assayed enzyme inhibitory activities, the extracts showed a significant antidiabetic effect in vitro with the highest activity corresponding to an IC50 of 3.88 µg/mL. The older oak bark represents a potential source of compounds with antibacterial, antioxidant or antidiabetic activity, which can be exploited through the biorefining processes.

Hypericum perforatum L., also known as St. John’s Wort, is recognized worldwide as a valuable medicinal herb; however, other Hypericum species were intensively studied for their bioactive potential. To fill the research gap that exists in the scientific literature, a comparative evaluation between H. alpigenum Kit., H. perforatum L. and H. rochelii Griseb. & Schenk was conducted in the present study. Two types of herbal preparations obtained from the aerial parts of these species were analyzed: extracts obtained through maceration and extracts obtained through magnetic-stirring-assisted extraction. LC-DAD-ESI-MSn analysis revealed the presence of phenolic acids, flavan-3-ols and flavonoid derivatives as the main constituents of the above-mentioned species. Moreover, all extracts were tested for their antioxidant, enzyme-inhibitory and antimicrobial potential. Our work emphasizes for the first time a detailed description of H. rochelii phenolic fractions, including their phytochemical and bioactive characterization. In comparison with the other two studied species, H. rochelii was found as a rich source of phenolic acids and myricetin derivatives, showing important antioxidant, anticholinesterase and antibacterial activity. The study offers new perspectives regarding the chemical and bioactive profile of the less-studied species H. alpigenum and H. rochelii.

Cherry stems (CS) represent a by-product intensively used in Eastern European countries as a traditional remedy for urinary tract disorders. Ethnopharmacological evidences sustain the use of CS as aqueous preparations (infusion and decoction), but few data were previously reported about phytochemical profile and pharmacological potential of CS hydroalcoholic extracts. In this regard, we aimed to evaluate the phenolic profile, in vitro antioxidant and tyrosinase inhibitory potential, and in vivo diuretic activity of 70% hydroethanolic cherry stems extract and cherry stems decoction (CSD). LC-DAD-ESI/MSn analysis revealed the presence of flavonoid-type compounds as main constituents for both preparations, especially flavanones (naringenin glycosides). Antioxidant activity evaluated through DPPH, ABTS, and FRAP methods was superior for cherry stems extract, probably due to the presence of phenolic-derived compounds in higher amounts than CSD. On the other hand, tyrosinase inhibitory potential and diuretic effect exerted by CSD were stronger, highlighting that other types of hydrophilic secondary metabolites are responsible for this bioactivity. Overall, our findings indicate that CS preparations could be used as promising mild diuretic agents and encourage further investigations regarding the correlation between their chemical composition and bioactive potential.

Acquired drug resistance represents a major clinical problem and one of the biggest limitations of chemotherapeutic regimens in colorectal cancer. Combination regimens using standard chemotherapeutic agents, together with bioactive natural compounds derived from diet or plants, may be one of the most valuable strategies to overcome drug resistance and re-sensitize chemoresistant cells. In this review, we highlight the effect of combined regimens based on conventional chemotherapeutics in conjunction with well-tolerated plant-derived bioactive compounds, mainly curcumin, resveratrol, and EGCG, with emphasis on the molecular mechanisms associated with the acquired drug resistance.

Snowdrop is an iconic early spring flowering plant of the genus Galanthus (Amaryllidaceae). Galanthus species (Galanthus spp.) are economically important plants as ornaments. Galanthus spp has gained significance scientific and commercial interest due to the discovery of Galanthamine as symptomatic treatment drug for Alzhiermer disease. This review aims to discuss the bioactivities of Galanthus spp including anticholinesterase, antimicrobial, antioxidant and anticancer potential of the extracts and chemical constituents of Galanthus spp. This review highlights that Galanthus spp. as the exciting sources for drug discovery and nutraceutical development.

Two techniques, namely, optimized ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE), were used to promote the extraction of phenolic compounds from the pseudo-fruits of Rosa canina L. (RC). For UAE, an optimization process based on the design of experiment (DoE) principles was used for determining the dependence between three variables (i.e., time of extraction, ultrasound amplitude, and the material-to-water ratio) and the total phenolic content of the samples. For EAE, a 2:1:1 pectinase, cellulase, and hemicellulase enzymatic blend was used as pre-treatment for optimized UAE, inducing a higher total phenolic content. The untargeted phenolic profiling approach revealed a great abundance of lower molecular weight phenolics (1.64 mg Eq./g) in UAE-RC extracts, whilst gallic acid (belonging to hydroxybenzoic acid derivatives) was the most abundant individual compound of both extracts. The unsupervised multivariate statistics clearly discriminated the impact of enzymatic pre-treatment on the phenolic profile of RC pseudo-fruits. Finally, Pearson’s correlation coefficients showed that anthocyanins, phenolic acids, and tyrosol derivatives were those compounds mostly correlated to the in vitro antioxidant potential of the extracts, whilst negative and significant (p < 0.05) correlation coefficients were recorded when considering the enzymatic inhibition activities. The highest enzyme-inhibitory activity has been identified against α-glucosidase, which indicates an antidiabetic effect.

Fumaria schleicheri Soy. Will. is a species belonging to the Papaveraceae family, being widespread in East-Central and Southern Europe. As with numerous other species of the genus, it is used in traditional medicine for the treatment of hepatobiliary and digestive disorders. The aim of the present study consisted of the evaluation of its alkaloid content and the assessment of its in vitro antioxidant, anti-cholinesterase and cytotoxic potential. Total alkaloid content in the composition of the species was quantified by a spectrophotometrical method and they were individually identified and quantified by HPLC-DAD. The antioxidant capacity was investigated by the DPPH and FRAP methods, while the anti-cholinesterase activity was assessed by an adapted Ellman’s method. The in vitro cytotoxic activity was evaluated on BJ human fibroblasts and DLD-1 human colon adenocarcinoma cell lines. Results showed the presence of bicuculline, protopine, chelidonine, stylopine and sanguinarine, among which bicuculline, protopine, stylopine and sanguinarine were quantified, while the antioxidant and anti-cholinesterase assays showed valuable potentials. No cytotoxic effect was observed on BJ cell lines and selective cytotoxicity was expressed towards tumoral cells. In this context, F. schleicheri appears as an important medicinal species with significant potential of substitution with the officinal species.

Malolactic fermentation (MLF) or biological decrease of wine acidity is defined as the enzymatic bioconversion of malic acid in lactic acid, a process performed by lactic acid bacteria (LAB). The procedures for the isolation of new indigenous LAB strains from the red wines produced in Copou Iasi wine center (NE of Romania) undergoing spontaneous malolactic fermentation, resulted in the obtaining of 67 catalase-negative and Gram-positive LAB strains. After testing in the malolactic fermentative process, application of specific screening procedures and identification (API 50 CH), two bacterial strains belonging to the species Oenococcus oeni (strain 13-7) and Lactobacillus plantarum (strain R1-1) with high yield of malolactic bioconversion, non-producing biogenic amines, and with active extracellular enzymes related to wine aroma, were retained and characterized. Tested in synthetic medium (MRS-TJ) for 10 days, the new isolated LAB strains metabolized over 98% of the malic acid at ethanol concentrations between 10 and 14 % (v/v), low pH (>3.0), total SO2 doses up to 70 mg/L and temperatures between 15 and 35 °C, showing high potential for future use in the winemaking process as bacterial starter cultures, in order to obtain high quality wines with increased typicity.

The species of the Quercus genus, including Quercus cerris L., are widely used and their wood represents a valuable material utilized for various purposes. The intense industrial processing of wood results in a considerable amount of poorly-used natural by-products, such as bark, and a loss of potentially useful raw materials. Thus, the aim of our study was to evaluate the phytochemical characteristics and potential biological activities of Quercus cerris bark extracts obtained by optimizing the parameters of microwave assisted extraction (MAE). The optimum conditions for MAE were determined using a design of experiments (DoE) model, which set the experimental variables (irradiation time and microwave power) and their values. Aqueous and hydroalcoholic extractions were performed and the optimum parameters of extraction were determined for both solvents. The total polyphenolic and tannin contents were determined. The biological activities representative of antioxidant capacity were determined using two free radical scavenging methods, the DPPH and ABTS methods, and the antibacterial activity was assessed with the microdilution method. The results showed different optimal extraction conditions for aqueous (30 min at 850 W) and hydroalcoholic (18 min at 650 W) extracts. A higher yield of total polyphenols was observed in the hydroalcoholic bark extract (403.73 ± 7.35 mg gallic acid equivalents/g dried weight); however a lower level of tannins was registered in comparison to the aqueous extract. In addition, both extracts exerted high antioxidant activities, with the aqueous extracts having a stronger inhibitory effect against the DPPH radical. Moreover, the extracts exhibited antibacterial activity against the tested bacterial strains, especially against the Gram-positive strains and Klebsiella pneumoniae, with the hydroalcoholic extracts being more efficient overall. To conclude, the optimized MAE was an efficient method to extract phytochemical compounds with potential biological effects from Quercus cerris bark.

Abstract Downy mildew of pearl millet caused by the biotrophic oomycete Sclerospora graminicola is the most devastating disease which impairs pearl millet production causing huge yield and monetary losses. Chitosan nanoparticles (CNP) were synthesized from low molecular weight chitosan having higher degree of acetylation was evaluated for their efficacy against downy mildew disease of pearl millet caused by Sclerospora graminicola. Laboratory studies showed that CNP seed treatment significantly enhanced pearl millet seed germination percentage and seedling vigor compared to the control. Seed treatment with CNP induced systemic and durable resistance and showed significant downy mildew protection under greenhouse conditions in comparison to the untreated control. Seed treatment with CNP showed changes in gene expression profiles wherein expression of genes of phenylalanine ammonia lyase, peroxidase, polyphenoloxidase, catalase and superoxide dismutase were highly upregulated. CNP treatment resulted in earlier and higher expression of the pathogenesis related proteins PR1 and PR5. Downy mildew protective effect offered by CNP was found to be modulated by nitric oxide and treatment with CNP along with NO inhibitors cPTIO completely abolished the gene expression of defense enzymes and PR proteins. Further, comparative analysis of CNP with Chitosan revealed that the very small dosage of CNP performed at par with recommended dose of Chitosan for downy mildew management.

Hot air drying has proven to be an efficient method to preserve specific edible plant materials with medicinal properties. This is a process involving chemical, physical, and biological changes in plant matrices. Understanding these processes will lead to an improvement in the yields of bioactive compounds. This study aims to optimize the drying process of two species’ fruits used in folk medicine, Berberis vulgaris and Crataegus monogyna. The optimized extracts’ antioxidant capacity was assessed using various assays, with the barberry extract showing very good activity (50.85, 30.98, and 302.45 mg TE/g dw for DPPH, TEAC, and FRAP assays, respectively). Both species exerted good fungal α-glucosidase inhibitory activity (IC50 = 0.34 and 0.56 mg/mL, respectively) but no activity on mammalian α-glucosidase. Additionally, this study identified and quantified the main bioactive compounds. The results presented herein are a breakthrough in industrializing this drying process. Additional studies are necessary to mechanistically understand the drying process involved in these plant materials.

High-resolution mass spectrometry is currently used to determine the mass of biologically active compounds in medicinal plants and food and UHPLC-Orbitrap is a relatively new technology that allows fast fingerprinting and metabolomics analysis. Forty-two metabolites including several phenolic acids, flavonoids, coumarines, tremetones and ent-clerodane diterpenes were accurately identified for the first time in the resin of the medicinal plant Parastrephia lucida (Asteraceae) a Chilean native species, commonly called umatola, collected in the pre-cordillera and altiplano regions of northern Chile, by means of UHPLC-PDA-HR-MS. This could be possible by the state of the art technology employed, which allowed well resolved total ion current peaks and the proposal of some biosynthetic relationships between the compounds detected. Some mayor compounds were also isolated using HSCCC. The ethanolic extract showed high total polyphenols content and significant antioxidant capacity. Furthermore, several biological assays were performed that determined the high antioxidant capacity found for the mayor compound isolated from the plant, 11- p-coumaroyloxyltremetone.

In this study, the nutraceutical potential of Capparis spinosa L. for the treatment of hyperglycemic states has been thoroughly investigated. A series of in vivo and in vitro tests have been conducted on fresh leaf, buds and salty buds (24 h desalted) processed to dry powder. 60% MeOH/H2O extracts were obtained for HPLC analysis and for α-amylase and α-glucosidase inhibition tests. To estimate the in vivo anti-diabetic effect, dry powders of C. spinosa leaf and buds were orally administered to streptozocin-induced diabetic rats over a period of 28 days. At the end of the experiment, animals were sacrificed, blood taken for assessment of lipid profile and liver/kidney biochemistry while section of the pancreas, liver and kidneys were processed for general histology. Results showed that the regular administration of C. spinosa leaf or buds normalized all the biochemical parameters and reversed the liver/kidney injury with variable degrees of organ protection.

Excessive alcohol consumption is the cause of several diseases and thus is of a major concern for society. Worldwide alcohol consumption has increased by many folds over the past decades. This urgently calls for intervention and relapse counteract measures. Modern pharmacological solutions induce complete alcohol self-restraint and prevent relapse, but they have many side effects. Natural products are most promising as they cause fewer adverse effects. Here we discuss in detail the medicinal plants used in various traditional/folklore medicine systems for targeting alcohol abuse. We also comprehensively describe preclinical and clinical studies done on some of these plants along with the possible mechanisms of action.

Sideritis, also named “ironwort,” “mountain tea,” or “shepherd's tea,” is a genus of flowering plants used as herbal medicine in traditional Mediterranean-area medicine systems, and these plants are generally consumed as a herbal tea. Its use as herbal tea and in traditional herbal medicine is quite popular. There are currently few studies on Sideritis perfoliata L., and only one reports the use of a liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MSn) profile and the content of phenolic compounds without considering a possible correlation with its biological activities. This paper aims to investigate the antioxidant activities by means of several different biological/biochemical assays (radical scavenging, reducing power, ferrous ion chelating, and total antioxidant by phosphomolybdenum and β-carotene bleaching methods) as well as analyze the enzyme inhibitory activities (against AChE (acetylcholinesterase), BChE (butyrylcholinesterase), tyrosinase, α-glucosidase, and α-amylase) as well as the total phenolics, flavonoids, and condensed tannins. The reported results on Sideritis perfoliata highlighted that methanol and water extracts generally showed higher radical scavenging and reducing power activities. A similar trend could be observed for phosphomolybdenum and ferrous ion chelating activities. Methanol extracts showed lower activity only for the β-carotene bleaching assay.

Arctium species are known for a variety of pharmacological effects due to their diverse volatile and non-volatile secondary metabolites. Representatives of Arctium species contain non-volatile compounds including lignans, fatty acids, acetylenic compounds, phytosterols, polysaccharides, caffeoylquinic acid derivatives, flavonoids, terpenes/terpenoids and volatile compounds such as hydrocarbons, aldehydes, methoxypyrazines, carboxylic and fatty acids, monoterpenes and sesquiterpenes. Arctium species also possess bioactive properties such as anti-cancer, anti-diabetic, anti-oxidant, hepatoprotective, gastroprotective, antibacterial, antiviral, antimicrobial, anti-allergic, and anti-inflammatory effects. This review aims to provide a complete overview of the chemistry and biological activities of the secondary metabolites found in therapeutically used Arctium species. Summary of pharmacopeias and monographs contents indicating the relevant phytochemicals and therapeutic effects are also discussed, along with possible safety considerations.

Cataract is the leading reason of blindness worldwide and is defined by the presence of any lens opacities or loss of transparency. The most common symptoms of cataract are impaired vision, decreased contrast sensitivity, color disturbance, and glare. Oxidative stress is among the main mechanisms involved in the development of age-related cataract. Surgery through phacoemulsification and intraocular lens implantation is the most effective method for cataract treatment, however, there are chances of serious complications and irreversible loss of vision associated with the surgery. Natural compounds consisting of antioxidant or anti-inflammatory secondary metabolites can serve as potential leads for anticataract agents. In this review, we tried to document medicinal plants and plant-based natural products used for cataract treatment worldwide, which are gathered from available ethnopharmacological/ethnobotanical data. We have extensively explored a number of recognized databases like Scifinder, PubMed, Science Direct, Google Scholar, and Scopus by using keywords and phrases such as “cataract”, “blindness”, “traditional medicine”, “ethnopharmacology”, “ethnobotany”, “herbs”, “medicinal plants”, or other relevant terms, and summarized the plants/phytoconstituents that are evaluated in different models of cataract and also tabulated 44 plants that are traditionally used in cataract in various folklore medical practices. Moreover, we also categorized the plants according to scientific studies carried out in different cataract models with their mechanisms of action.

Backround:Ajuga species have been used in traditional medicine for their diuretic, anti-inflammatory, wound-healing, and hepatoprotective properties.Purpose: The phytochemical profile and anticancer potential of three Ajuga sp. (A. genevensis, A. chamaepitys, and A. laxmannii) from Romania was investigated.Materials and Methods: The phytochemicals were extracted from the aerial parts of Ajuga sp. by using different solvents and methods. The hydroalcoholic extracts were examined for total phenolic, flavonoid and iridoid contents, and HPLC/MS was used to analyze the polyphenolic compounds and iridoids. The phytochemical profile was also evaluated by principal component analysis in connection with antitumor efficacy of extracts. The antiproliferative potential was evaluated using the ELISA BrdU-colorimetric immunoassay. Western Blot with regard to inflammatory protein NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) p65 subunit expression in cell lysates was performed. Quantification of oxidative stress marker malondialdehyde (MDA) was determined by high-performance liquid chromatography (HPLC). Enzymatic and non-enzymatic antioxidant capability was assessed by measuring catalase activity and by evaluating the total antioxidant capacity (TAC) of treated cells.Results:Ajuga laxmannii ethanol extract showed the highest total phenolic and flavonoid content, while A. genevensis ethanol extract was more abundant in iridoids. The overall cytostatic effect of the investigated plant extracts was exerted through strong inhibitory actions on NF-κB, the key molecule involved in the inflammatory response and via oxidative stress modulatory effects in both murine colon carcinoma and melanoma cell lines.Conclusion:Ajuga laxmannii showed the most significant antitumor activity and represents an important source of bioactive compounds, possibly an additional form of treatment alongside conventional anticancer drugs.

Prunus mahaleb L. fruit has long been used in the production of traditional liqueurs. The fruit also displayed scavenging and reducing activity, in vitro. The present study focused on unravelling peripheral and central protective effects, antimicrobial but also anti-COVID-19 properties exerted by the water extract of P. mahaleb. Anti-inflammatory effects were studied in isolated mouse colons exposed to lipopolysaccharide. Neuroprotection, measured as a blunting effect on hydrogen-peroxide-induced dopamine turnover, was investigated in hypothalamic HypoE22 cells. Antimicrobial effects were tested against different Gram+ and Gram- bacterial strains. Whereas anti-COVID-19 activity was studied in lung adenocarcinoma H1299 cells, where the gene expression of ACE2 and TMPRSS2 was measured after extract treatment. The bacteriostatic effects induced on Gram+ and Gram- strains, together with the inhibition of COX-2, TNFα, HIF1α, and VEGFA in the colon, suggest the potential of P. mahaleb water extract in contrasting the clinical symptoms related to ulcerative colitis. The inhibition of the hydrogen peroxide-induced DOPAC/DA ratio indicates promising neuroprotective effects. Finally, the downregulation of the gene expression of ACE2 and TMPRSS2 in H1299 cells, suggests the potential to inhibit SARS-CoV-2 virus entry in the human host. Overall, the results support the valorization of the local cultivation of P. mahaleb.

Goji (Lycium barbarum L.) leaves are emphasized as a functional tea or as dietary supplements. The phenolic compound profile, antioxidant, enzyme inhibitory, antimicrobial, and antimutagenic activities of leaf extracts from two selected cultivars in comparison with wild-growing plants have been evaluated. HPLC-DAD/ESI-ToF-MS analysis revealed the presence of phenolic acids and flavonoids with chlorogenic acid and rutin being the dominant compounds in the cultivated plants, whereas rutin and kaempeferol-3-O-rutinoside for wild growing ones. In particular, cv. Erma contained the highest amount of chlorogenic acid and showed a strong tyrosinase-inhibitory effect. Staphylococcus aureus, Listeria monocytogenes, and Penicillium funiculosum were the most sensitive strains when exposed to extracts from cultivated plants. Antimutagenic activity was evaluated by Ames' test. The tested extracts provided high protection against mutagenicity induced by 2-anthramine (2-AA) to Salmonella typhimurium strains TA 98 and TA 100 (max. inhibition (%) 88% and 74.2%, respectively). Overall, Goji leaves are a rich source of bioactive compounds with functional properties that need further risk/benefit evaluation when used in foods or health-promoting formulations.

Morina persica L. has weak rosy smell flowers that together with aerial parts are infused or decocted and used as a functional tea. It is a representative of the Anatolian traditional medicine and is used for the treatment of cold. Biological and chemical fingerprints of different extracts (methanolic, acetone and water) of M. persica were investigated. Antioxidant, enzyme inhibitory, antimicrobial, cytotoxic and mutagenic/antimutagenic effects were evaluated for biological profiling. Rutin, chlorogenic and rosmarinic acids were detected as dominant compounds. Generally, the methanolic and acetone extracts had strong antioxidant, enzyme inhibitory, antimicrobial and antimutagenic effects. Additionally, all extracts showed remarkable inhibitory effects on HeLa cells at 100 µg/mL. Furthermore, possible interactions between rutin, the dominant phenolic compound in M. persica and the tested enzymes were assessed by molecular docking. Results indicate that M. persica could be considered as a natural source of high-valued functional ingredients for further use in healthful formulations.

Schisandra chinensis is a widely-consumed nutraceutical providing advantageous nutritional and bioactive properties. This study aimed at characterizing the chemical composition and antioxidant capacity of S. chinensis fruits as well as the underestimated leaves and stems. A special aim of this study was to characterize the contribution of single constituents (lignans and further phenolic compounds) to the overall antioxidant activity using an HPLC-online TEAC approach. By using LC-DAD/ESI-ToF-MS, twenty-eight lignans and twenty phenolic compounds were identified. Among the 48 characterized components, twelve phenolic compounds were identified for the first time in Schisandra. Gomisin D was the only lignan being able to scavenge ABTS+ radicals, suggesting that not its amount but rather its chemical structure is responsible. Overall, clorogenic acid isomers and quercetin glycosides contribute to over 80% of the total antioxidant activity. This study indicates that leaves and stems of S. chinensis may be used for obtaining bioactive formulations and functional foods.

The chemical composition and biological properties correlation in several medicinal and aromatic plants is still underexplored, especially in its most common form of consumption as tisane. The present study aims to characterize the organic acids and vitamin E composition of five tisanes and their extracts by High-Performance Liquid Chromatography coupled to a diode-array detector (HPLC-DAD) and HPLC coupled to a fluorescence detector techniques, respectively, and the phenolic composition by HPLC-DAD-ESI/MS (mass spectrometry by electrospray ionization). It also focuses on their bioactive properties, namely antioxidant, antimicrobial, anti-inflammatory, cytotoxic, anti-tyrosinase, and anti-diabetic activities. A Principal Component Analysis (PCA) was performed in order to understand the correlation between the chemical composition and bioactive properties of the tisanes. The tisane 5 (T5) composed by lemon thyme, tutsan, cloves, and cinnamon, was the most promising mixture, presenting the lowest values for the lipid peroxidation inhibition, anti-inflammatory, and anti-diabetic activity. It also presented the highest concentration of phenolic acids (caffeoylquinic acids derivatives), and flavan-3-ols (catechin derivatives). Only the dry plants presented tocopherols. For the antihemolytic, antimicrobial, and cytotoxic activity, T2 and T4 (with lemon thyme) were highlighted as the best herbal mixtures. The PCA proved to be a valid tool to select the most promising tisane according to the bioactivity. These results suggest that the studied tisanes can be source of high added-value bioactive compounds with health-promoting effects and potential for application in the food and nutraceutical industries, among others.

The concern for implementing bioactive nutraceuticals in antioxidant-related therapies is of great importance for skin homeostasis in benign or malignant diseases. In order to elucidate some novel insights of Lycium barbarum (Goji berry) activity on skin cells, the present study focused on its active compound zeaxanthin. By targeting the stemness markers CD44 and CD105, with deep implications in skin oxidative stress mechanisms, we revealed, for the first time, selectivity in zeaxanthin activity. When applied in vitro on BJ human fibroblast cell line versus the A375 malignant melanoma cells, despite the moderate cytotoxicity, the zeaxanthin-rich extracts 1 and 2 were able to downregulate significantly the CD44 and CD105 membrane expression and extracellular secretion in A375, and to upregulate them in BJ cells. At mechanistic level, the present study is the first to demonstrate that the zeaxanthin-rich Goji extracts are able to influence selectively the mitogen-activated protein kinases (MAPK): ERK, JNK and p38 in normal BJ versus tumor-derived A375 skin cells. These results point out towards the applications of zeaxanthin from L. barbarum as a cytoprotective agent in normal skin and raises questions about its use as an antitumor prodrug alone or in combination with standard therapy.

Blueberries have seen an ascending production line boosted by World Health Organization (WHO) approvals for their contributions to a healthy diet and the evidence that they act against different diseases. This increase resulted in significant amounts of discarded leaves, which could be a valuable source of bioactive compounds. In the present study, ultrasound-assisted extraction technology was used to determine and compare the chemical and biological profiles of leaves from six commercial blueberry (Vaccinium corymbosum L.) varieties. Feruloylquinic acid was the major compound identified, ranging from 19.23 ± 0.18 mg/g (at the lowest level, registered in the Spartan variety) to 49.62 ± 0.41 mg/g (at the highest level, registered in the Nelson variety). Rutin was the second major compound identified, for which Toro, Nelson, and Elliot leaves registered the highest values, with 35.77 ± 0.19 mg/g, 32.50 ± 0.20 mg/g, and 31.53 ± 0.1 mg/g, respectively. Even though analogous polyphenols were detected in the six cultivars, their concentrations and amounts were different. The leaf extracts of the cultivars Toro, Elliot, and Nelson appear to be good sources of antioxidants, registering high percentage inhibitions of DPPH radicals, of 70.41%, 68.42%, and 58.69%, respectively. The blueberry leaf extracts had a strong antibacterial activity and a low antifungal capacity, and a low-to-moderate antimutagenic capacity towards Salmonella typhimurium TA98 and TA100 strains, with Toro leaf being the best candidate. All of these biological activities indicate health-related benefits, recommending them as suitable candidates for medical and pharmaceutical applications. The present paper adds significant knowledge to the field of blueberry leaves via chemical and biological profiles, supporting the ultrasound-assisted extraction technique as a useful and green method to provide alternative sources of bioactive compounds.

Strawberries belonging to cultivar Clery (Fragaria x ananassa (Duchesne ex Weston)), cultivated in central Italy were subjected to a multi-methodological experimental study. Fresh and defrosted strawberries were exposed to different processing methods, such as homogenization, thermal and microwave treatments. The homogenate samples were submitted to CIEL*a*b* color analysis and Head-Space GC/MS analysis to determine the impact of these procedures on phytochemical composition. Furthermore, the corresponding strawberry hydroalcoholic extracts were further analyzed by HPLC-DAD for secondary metabolites quantification and by means of spectrophotometric in vitro assays to evaluate their total phenolic and total flavonoid contents and antioxidant activity. These chemical investigations confirmed the richness in bioactive metabolites supporting the extraordinary healthy potential of this fruit as a food ingredient, as well as functional food, highlighting the strong influence of the processing steps which could negatively impact on the polyphenol composition. Despite a more brilliant red color and aroma preservation, non-pasteurized samples were characterized by a lower content of polyphenols and antioxidant activity with respect to pasteurized samples, as also suggested by the PCA analysis of the collected data.

Even though Salvia genus is one of the most known and studied taxa of Lamiaceae family, the knowledge regarding the chemical composition and health-related benefits of some locally used Salvia species (mostly endemic) is still scarce. In this regard, the present work aims to evaluate the chemical profile and potential bioactivities of 70% (v/v) ethanolic extracts obtained from the less-studied S. transsylvanica and S. glutinosa in comparison with S. officinalis. HPLC-PDA analysis revealed the presence of rutin and catechin as the main compounds in the extracts of the three studied species (using the employed HPLC method), whereas the presence of naringenin was highlighted only in S. glutinosa extract. Chlorogenic acid, rutin and quercetin were identified and quantified for the first time in S. transsylvanica extracts. The in vitro antioxidant capacity of each extract was tested through complementary methods (phosphomolybdenum assay, DPPH, ABTS, CUPRAC and FRAP assays), and correlated with the presence of phenolics (especially flavonoids) in high amounts. The neuroprotective and antidiabetic abilities of S. officinalis (the most active as AChE, BChE and α-glucosidase inhibitor), S. glutinosa (the most active as α-amylase inhibitor) and S. transsylvanica were also studied. For each extract it was determined the antimicrobial, antifungal and cytotoxic effects using in vitro assays. The obtained results confirm the potential of S. transsylvanica and S. glutinosa as promising sources of bioactive compounds and as a starting point for further analyses.

This study was performed to evaluate and compare the in vitro antioxidant, antimicrobial, and antimutagenic activities, and the polyphenolic content of the Nord-West Romanian wild bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.) leaves from three different natural habitats (Smida, Turda, Borsa). In the case of both species, the flavanols level was higher in Smida habitat (altitude 1100 m), whereas quercetin derivates were more abundant in Borsa habitat (altitude 850 m). The bilberry leaf extracts contained in the highest amounts the feruloylquinic acid (59.65 ± 0.44 mg/g for Borsa habitat) and rutin (49.83 ± 0.63 mg/g for Borsa habitat), and showed relevant 2,2-diphenyl-1-picrylhydrazyl (DPPH) antioxidant activity (271.65 mM Trolox/100 g plant material for Borsa habitat, 262.77 mM Trolox/100 g plant material for Smida habitat, and 320.83 mM Trolox/100 g plant material for Turda habitat), for all the three extracts. Gallocatechin was the dominant flavanol in lingonberry species, with the highest amount being registered for Smida habitat (46.81 ± 0.3 mg/g), revealing a DPPH antioxidant activity of 251.49 mM Trolox/100 g plant material. The results obtained in the antimicrobial tests showed that the best inhibitory effect among bilberry species was attributed to the Turda (altitude 436 m) and Smida locations, against both Gram-positive and Gram-negative bacterial strains. For lingonberry, the differences in habitat did not influence the antibacterial effect, but the antifungal effect, only in the case of Candida zeylanoides. A strong antimutagenic effect was registered by the bilberry leaves toward Salmonella typhimurium TA100. Our study may be able to provide a better understanding of the correlation between natural habitat conditions and the accumulation of secondary metabolites and their related bioactivities in studied leaves.

Walnut (Juglans regia L.) septum represents an interesting bioactive compound source by-product. In our study, a rich phenolic walnut septum extract, previously selected, was further examined. The tocopherol content determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed higher amounts of α-tocopherol compared to γ- and δ-tocopherols. Moreover, several biological activities were investigated. The in vitro inhibiting assessment against acetylcholinesterase, α-glucosidase, or lipase attested a real management potential in diabetes or obesity. The extract demonstrated very strong antimicrobial potential against Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella enteritidis. It also revealed moderate (36.08%) and strong (43.27%) antimutagenic inhibitory effects against TA 98 and TA 100 strains. The cytotoxicity of the extract was assessed on cancerous (A549, T47D-KBluc, MCF-7) and normal (human gingival fibroblasts (HGF)) cell lines. Flow cytometry measurements confirmed the cytotoxicity of the extract in the cancerous cell lines. Additionally, the extract demonstrated antioxidant activity on all four cell types, as well as anti-inflammatory activity by lowering the inflammatory cytokines (interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-1 β (IL-1β)) evaluated in HGF cells. To the best of our knowledge, most of the cellular model analyses were performed for the first time in this matrix. The results prove that walnut septum may be a potential phytochemical source for pharmaceutical and food industry.

Antioxidant dietary intervention is considered a potential strategy in delaying age-related dysfunctions. In this study of 56 days, we assessed the antioxidant effects of walnut kernel (WK) and walnut septum extract (WSE) in a D-galactose (D-gal)-induced aging model and in a naturally aged rat model. Young Wistar rats, treated with D-gal (1200 mg/week), and old rats received daily WK or WSE added to the feed. After 8 weeks, blood, liver, and brain samples were collected and hematological, biochemical, oxidative stress biomarkers, histological, and immunohistochemical analyses were performed. Moreover, acetylcholinesterase activity was investigated in brain homogenates. The outcomes demonstrated significant improvement in cellular antioxidant activity and/or decrease of reactive oxygen species, advanced glycation end products, nitric oxide, malondialdehyde, or increase of glutathione after WK or WSE intake in both models. Additionally, WSE showed hypoglycemic effect, and both WK and WSE lowered acetylcholinesterase activity. Both diets could protect neurons against the induced senescence and could reverse the pathological conditions in the physiological aged brain. Thus, dietary supplementation with WK or WSE can maintain the liver and brain health and reduce the risk of age-related diseases, as well as delaying the onset of aging processes.