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Aquilegia pubiflora leaves are reported against hepatitis, for treatment of skin burns, wound healing, jaundice, gynea, circulatory and cardiovascular disease in several studies. Therefore, green synthesis of silver nanoparticles with A. pubiflora leaves extract, as an alternative to chemical synthesis, is beneficial from its biological applications point of view. The formation and size distribution of Ag-NPs were confirmed by UV-spectroscopy, Scanning electron microscopy (SEM), X-ray powder diffraction (XRD), Transmission electron microscopy (TEM), Energy Dispersive X-ray (EDX), Fourier transform infrared spectroscopy (FTIR) and HPLC. Silver nanoparticles with a particle size of 19 nm were used in various assays including antimicrobial, antioxidant, anti-Alzheimer, anti-diabetic, anti-aging, anti-cancer and biocompatibility studies. Our findings revealed that A. pubiflora mediated Ag-NPs were highly active against both bacterial and fungal strains with Escherichia coli and Aspergillus niger displaying the highest zone of inhibition 10.9 ± 0.29 mm and 13.11 ± 0.33 mm respectively. Dose dependent cytotoxic activity was shown by NPs against Leishmania Tropica with significant IC50 of 175 μg/mL for promastigote and 169 μg/mL for amastigote forms. The Ag-NPs proved to be highly toxic for the HepG2 cell line and inhibit cell viability up to 52.06 ± 2.9% after 24 h of Ag-NPs treatment. Ag-NPs also showed excellent inhibitory potential against the enzymes acetylcholinesterase (AChE: 71.38 ± 1.75) and butyrylcholinesterase (BChE: 73.61 ± 1.96) which are involved in Alzheimer's disease. Overall, the enzymes involved in aging, diabetes, and inflammation showed a moderate inhibitory response to Ag-NPs, led to the conclusion that they are a promising choice for various biological applications.

Abstract Ajuga integrifolia Buch. Ham. ex D.Don, a member of Lamiaceae family is pharmaceutically an active perennial herb widely spread in China, Afghanistan and Pakistan Himalayan region. The application of biotic elicitors is a promising approach to cover limitations of in vitro cell technology and challenges faced by pharmaceuticals industry for bulk up production. The current study involved the induction of agitated micro-shoot cultures with the aim to investigate the growth-promoting as well as phytochemicals enhancement role of yeast extract (YE) and pectin (PE). The results showed that both elicitors induced a considerable physiological response. Biomass accumulation was observed maximum (DW: 18.3 g/L) against PE (10 mg/L) compared to YE and control. Eleven secondary phytocompounds were quantified using high-performance liquid chromatography. PE (50 mg/L) was found to be effective in elicitation of rosmarinic acid (680.20 µg/g), chlorogenic acid (294.12 µg/g), apigenin (579.61 µg/g) and quercetin (596.89 µg/g). However, maximum caffeic acid (359.52 µg/g) and luteolin (546.12 µg/g accumulation was noted in PE (1 mg/L) treatment. Harpagide, aucubin, harpagoside and 8-O-acetyl-harpagoside production was suppressed by both elicitors except for YE (100 mg/L). Catalpol accumulation in micro-shoot cultures was also downregulated except in response to YE (50 and 100 mg/L). Antioxidant activity and anti-inflammatory activity remained higher under PE (50 mg/L) and YE (100 mg/L) respectively. Therefore, results suggested that Ajuga integrifolia micro-shoot cultures treated with yeast extract and pectin might be an efficient bio-factory to produce commercially potent specific secondary metabolites.

Abstract Background Himalayan Columbine (Aquilegia pubiflora Wall. Ex Royle) is a medicinal plant and have been used as traditional treatments for various human diseases including skin burns, jaundice, hepatitis, wound healing, cardiovascular and circulatory diseases. Till now there is no report available on phytochemical investigation of Himalayan Columbine and to the best of our knowledge, through present study we have reported for the first time, the phytochemical analysis and pharmacological potentials of different leaf extracts of Aquilegia pubiflora. Methods Four types of extracts were prepared using solvent of different polarities (Distilled water APDW, Methanol APM, Ethanol APE and Ethyl acetate APEA), and were evaluated to determine the best candidate for potent bioactivity. Phytochemical constituents in prepared extracts were quantified through HPLC analysis. Subsequently, all four types of leaf extracts were then evaluated for their potential bioactivities including antimicrobial, protein kinase inhibition, anti-inflammatory, anti-diabetic, antioxidant, anti-Alzheimer, anti-aging and cytotoxic effect. Results HPLC analysis demonstrated the presence of dvitexin, isovitexin, orientin, isoorientin, ferulic acid, sinapic acid and chlorogenic acid in varied proportions in all plant extracts. Antimicrobial studies showed that, K. pneumonia was found to be most susceptible to inhibition zones of 11.2 ± 0.47, 13.9 ± 0.33, 12.7 ± 0.41, and 13.5 ± 0.62 measured at 5 mg/mL for APDW, APM, APE and APEA respectively. A. niger was the most susceptible strain in case of APDW with the highest zone of inhibition 14.3 ± 0.32, 13.2 ± 0.41 in case of APM, 13.7 ± 0.39 for APE while 15.4 ± 0.43 zone of inhibition was recorded in case of APEA at 5 mg/mL. The highest antioxidant activity of 92.6 ± 1.8 μgAAE/mg, 89.2 ± 2.4 μgAAE/mg, 277.5 ± 2.9 μM, 289.9 ± 1.74 μM for TAC, TRP, ABTS and FRAP, respectively, was shown by APE. APM, APE and APEA extracts showed a significant % cell inhibition (above 40%) against HepG2 cells. The highest anti-inflammatory of the samples was shown by APE (52.5 ± 1.1) against sPLA2, (41.2 ± 0.8) against 15-LOX, followed by (38.5 ± 1.5) and (32.4 ± 0.8) against COX-1 and COX-2, respectively. Conclusions Strong antimicrobial, Protein Kinase potency and considerable α-glucosidase, α-amylase, and cytotoxic potential were exhibited by plant samples. Significant anti-Alzheimer, anti-inflammatory, anti-aging, and kinase inhibitory potential of each plant sample thus aware us for further detailed research to determine novel drugs.

Abstract Fagonia indica (L.) is an important medicinal plant with multitude of therapeutic potentials. Such application has been attributed to the presence of various pharmacological important phytochemicals. However, the inadequate biosynthesis of such metabolites in intact plants has hampered scalable production. Thus, herein, we have established an in vitro based elicitation strategy to enhance such metabolites in callus culture of F. indica. Cultures were exposed to various doses of UV radiation (UV-C) and grown in different photoperiod regimes and their impact was evaluated on biomass accumulation, biosynthesis of phytochemicals along antioxidant expression. Cultures grown under photoperiod (16L/8D h) after exposure to UV-C (5.4 kJ/m2) accumulated optimal biomass (438.3 g/L FW; 16.4 g/L DW), phenolics contents (TPC: 11.8 μgGAE/mg) and flavonoids contents (TFC: 4.05 μgQE/mg). Similarly, HPLC quantification revealed that total production (6.967 μg/mg DW) of phytochemicals wherein kaempferol (1.377 μg/mg DW), apigenin (1.057 μg/mg DW), myricetin (1.022 μg/mg DW) and isorhamnetin (1.022 μg/mg DW) were recorded highly accumulated compounds in cultures at UV-C (5.4 kJ/m2) dose than other UV-C radiations and light regimes.. The antioxidants activities examined as DPPH (92.8%), FRAP (182.3 µM TEAC) and ABTS (489.1 µM TEAC) were also recorded highly expressed by cultures under photoperiod after treatment with UV-C dose 5.4 kJ/m2. Moreover, same cultures also expressed maximum % inhibition towards phospholipase A2 (sPLA2: 35.8%), lipoxygenase (15-LOX: 43.3%) and cyclooxygenases (COX-1: 55.3% and COX-2: 39.9%) with 1.0-, 1.3-, 1.3- and 2.8-fold increased levels as compared with control, respectively. Hence, findings suggest that light and UV can synergistically improve the metabolism of F. indica and could be used to produce such valuable metabolites on commercial scale.

Nano-elicitation is one among the prioritised strategies considered globally for sustainable and uniform production of industrially important medicinal compounds. Ocimum basilicum (Thai basil), a renowned medicinal species is a reservoir of commercially vital metabolites and proved for its health assuring effects in cancer, diabetes, microbial and cardiovascular diseases. However, its consumption and industrial demand raised intent to divert towards better alternates for ensuring sustainable production of medicinal compounds. Herein, we investigated the comparative potential of metal oxide [copper oxide (CuO) and manganese oxide (MnO)] nanoparticles to elicit the biosynthesis of bioactive metabolites and antioxidative capacity of O.basilicum callus cultures. Results showed that callus grown on MS media supplemented with 10 mg/L CuO-NPs resulted in the highest biomass accumulation (FW: 172.8 g/L, DW: 16.7 g/L), phenolic contents (TPC: 27.5 mg/g DW), and flavonoid contents (TFC: 9.1 mg/g DW) along with antioxidant activities (DPPH: 94%, ABTS: 881 μM TEAC, FRAP: 386 μM TEAC) compared with MnO-NPs and control. Likewise, the Superoxide dismutase (SOD: 1.28 nM/min/mg FW) and Peroxidase (POD: 0.48 nM/min/mg FW) activities were also recorded maximum in CuO-NPs elicited cultures than MnO-NPs and control. Moreover, the HPLC results showed that rosmarinic acid (11.4 mg/g DW), chicoric acid (16.6 mg/g DW), eugenol (0.21 mg/g DW) was found optimum in cultures at 10 mg/L CuO-NPs. Overall, it can be concluded that CuO nanoparticles can be effectively used as a elicitor for biosynthesis of metabolites in callus cultures of O. basilicum (Thai basil). The study is indeed a contribution to the field that will help decoding the mechanism of action of CuO NPs. However, further molecular investigations are needed to fully develop understanding about the metabolic potential of O. bascillicum and scalling up this protocol for bulkup production of bioactive compounds.

Rivea hypocrateriformis (Desr.) Choisy is a robust woody climbing shrub of the genus Rivea which is widely distributed in India, Nepal, Sri Lanka, Pakistan, Bangladesh, Myanmar, and Thailand. R. hypocrateriformis is a promising medicinal herb with a wide range of beneficial and health-promoting properties. Since the ancient times, it has been used as a traditional medicine to treat rheumatic pain, fever, urogenital problems, snake bites, cough, piles, malaria, and skin diseases. Aside from these traditional uses, its leaves and young shoots are also cooked and eaten as a vegetable and used for the preparation of bread with millet flour. This study extensively analyzes the available information on R. hypocrateriformis botanical characterization, distribution, traditional applications, phytochemistry, pharmacology, and toxicological properties. Phytochemical investigations of the plant has revealed the presence of highly valuable secondary metabolites including alkaloids, glycosides, coumarins, flavonoids, xanthones, stilbenes, and other organic compounds. Its crude extracts and isolated compounds have revealed anovulatory, antifertility, antiarthritic, antimicrobial, anticancer, antioxidant hepatoprotective, antilithiatic, and antimitotic potentials. This review of literature clearly identifies R. hypocrateriformis as a potent medicinal plant with remarkable healing and health-promoting properties. Further research directions into the bioactive extracts, clinical, and toxicological evaluations to assess the beneficial health-promoting properties of this promising herb are also discussed.

Stamen tea from Nelumbo nucifera Gaertn. (or the so-called sacred lotus) is widely consumed, and its flavonoids provide various human health benefits. The method used for tea preparation for consumption, namely the infusion time, may affect the levels of extractable flavonoids, ultimately affecting their biological effects. To date, there is no report on this critical information. Thus, this study aims to determine the kinetics of solid liquid extraction of flavonoid from sacred lotus stamen using the traditional method of preparing sacred lotus stamen tea. Phytochemical composition was also analyzed using high-performance liquid chromatography (HPLC). The antioxidant potential of stamen tea was also determined. The results indicated that the infusion time critically affects the concentrations of flavonoids and the antioxidant capacity of sacred lotus stamen tea, with a minimum infusion time of 5–12 min being required to release the different flavonoids from the tea. The extraction was modeled using second order kinetics. The rate of release was investigated by the glycosylation pattern, with flavonoid diglycosides, e.g., rutin and Kae-3-Rob, being released faster than flavonoid monoglycosides. The antioxidant activity was also highly correlated with flavonoid levels during infusion. Taken together, data obtained here underline that, among others, the infusion time should be considered for the experimental design of future epidemiological studies and/or clinical trials to reach the highest health benefits.

Nanotechnology is an emerging area of research that deals with the production, manipulation, and application of nanoscale materials. Bio-assisted synthesis is of particular interest nowadays, to overcome the limitations associated with the physical and chemical means. The aim of this study was to synthesize ZnO nanoparticles (NPs) for the first time, utilizing the seed extract of Lepidium sativum. The synthesized NPs were confirmed through various spectroscopy and imagining techniques, such as XRD, FTIR, HPLC, and SEM. The characterized NPs were then examined for various in vitro biological assays. Crystalline, hexagonal-structured NPs with an average particle size of 25.6 nm were obtained. Biosynthesized ZnO NPs exhibited potent antioxidant activities, effective α-amylase inhibition, moderate urease inhibition (56%), high lipase-inhibition (71%) activities, moderate cytotoxic potential, and significant antibacterial activity. Gene expression of caspase in HepG2 cells was enhanced along with elevated production of ROS/RNS, while membrane integrity was disturbed upon the exposure of NPs. Overall results indicated that bio-assisted ZnO NPs exhibit excellent biological potential and could be exploited for future biomedical applications. particularly in antimicrobial and cancer therapeutics. Moreover, this is the first comprehensive study on Lepidium sativum-mediated synthesis of ZnO nanoparticles and evaluation of their biological activities.

Nymphaea lotus L. is a potential plant in the Nymphaeaceae family that is well-recognized as an economic and traditional medicinal plant in Thailand and other countries. Its pharmacological and medicinal effects have been confirmed. However, there is no study going deeper into the population level to examine the phytochemical variation and biological activity of each population that benefits phytopharmaceutical and medical applications using this plant as raw material. This study was intensely conducted to complete this important research gap to investigate the flavonoid profiles from its floral parts, the stamen and perianth, as well as the antioxidant potential of the 13 populations collected from every floristic region by (1) analyzing their flavonoid profiles, including the HPLC analysis, and (2) investigating the antioxidant capacity of these populations using three assays to observe different antioxidant mechanisms. The results indicated that the northeastern and northern regions are the most abundant floristic regions, and flavonoids are the main phytochemical class of both stamen and perianth extracts from N. lotus. The stamen offers higher flavonoids and richer antioxidant potential compared with the perianth. This finding is also the first completed report at the population level to describe the significant correlation between the phytochemical profiles in floral parts extracts and the main antioxidant activity, which is mediated by the electron transfer mechanism. The results from the Pearson correlation coefficients between several phytochemicals and different antioxidant assessments highlighted that the antioxidant capability of these extracts is the result of complex phytochemical combinations. The frontier knowledge from these current findings helps to open up doors for phytopharmaceutical industries to discover their preferred populations and floral parts that fit with their targeted products.

Solanum virginianum L. (Solanum xanthocarpum) is an important therapeutic plant due to the presence of medicinally useful plant-derived compounds. S. virginianum has been shown to have anticancer, antioxidant, antibacterial, antiaging, and anti-inflammatory properties. This plant is becoming endangered due to overexploitation and the loss of its native habitat. The purpose of this research is to develop an ideal technique for the maximum biomass and phytochemical accumulation in S. virginianum leaf-induced in vitro cultures, as well as to evaluate their potential antiaging, anti-inflammatory, and antioxidant abilities. Leaf explants were grown on media (Murashige and Skoog (MS)) that were supplemented with various concentrations and combinations of plant hormones (TDZ, BAP, NAA, and TDZ + NAA) for this purpose. When compared with the other hormones, TDZ demonstrated the best response for callus induction, biomass accumulation, phytochemical synthesis, and biological activities. However, with 5 mg/L of TDZ, the optimal biomass production (FW: 251.48 g/L and DW: 13.59 g/L) was estimated. The highest total phenolic level (10.22 ± 0.44 mg/g DW) was found in 5 mg/L of TDZ, whereas the highest flavonoid contents (1.65 ± 0.11 mg/g DW) were found in 10 mg/L of TDZ. The results of the HPLC revealed that the highest production of coumarins (scopoletin: 4.34 ± 0.20 mg/g DW and esculetin: 0.87 ± 0.040 mg/g DW) was determined for 10 mg/L of TDZ, whereas the highest accumulations of caffeic acid (0.56 ± 0.021 mg/g DW) and methyl caffeate (18.62 ± 0.60 mg/g DW) were shown by 5 mg/L of TDZ. The determination of these phytochemicals (phenolics and coumarins) estimates that the results of our study on biological assays, such as antioxidant, anti-inflammatory, and antiaging assays, are useful for future cosmetic applications.

Grape canes represent a valuable source of numerous polyphenols with antioxidant properties, whose compositions vary depending on the genotype and environmental factors. Antioxidant activities of pure molecules are often reported without considering possible interactions that may occur in complex polyphenol mixture. Using UPLC-MS-based metabolomics and unsupervised classification, we explored the polyphenol variations in grape cane extracts from a collection of European varieties. Antioxidant activities were assessed using ORAC, ABTS, DPPH, FRAP, CUPRAC and chelation assays. Pairwise correlations between polyphenols and antioxidant capacities were performed to identify molecules that contributed more to the antioxidant capacities within a complex mixture of polyphenols.

Plants of the genus Monochoria have long been utilized in food, cosmetics, and traditional herbal treatments. Thailand has the highest species diversity of this genus and a new member, Monochoria angustifolia (G. X. Wang) Boonkerd & Tungmunnithum has been recently described. This plant is called “Siam Violet Pearl” as a common name or “Khimuk Si Muang Haeng Siam” as its vernacular name with the same meaning in the Thai language. Despite their importance, little research on Monochoria species has been conducted. This study, thus, provides the results to fill in this gap by: (i) determining flavonoid phytochemical profiles of 25 natural populations of M. angustifolia covering the whole floristic regions in Thailand, and (ii) determining antioxidant activity using various antioxidant assays to investigate probable mechanisms. The results revealed that M. angustifolia presents a higher flavonoid content than the outgroup, M. hastata. Our results also revealed that flavonoids might be used to investigate Monochoria evolutionary connections and for botanical authentication. The various antioxidant assays revealed that M. angustifolia extracts preferentially act through a hydrogen atom transfer antioxidant mechanism. Pearson correlation analysis indicated significant correlations, emphasizing that the antioxidant capacity is most probably due to the complex action of several phytochemicals rather than that of a single molecule. Together, these results showed that this new species provide an attractive alternative starting material with phytochemical variety and antioxidant potential for the phytopharmaceutical industry.

Punica granatum is a tree of the Punicaceae family which is widespread all over the world with several types of varieties. Its fruit juice is highly prized, whereas the bark, rich in in phytochemicals such as flavonoids, hydrolysable tannins, phenolic acids, and fatty acids, is regarded an agro-industrial waste. It is utilized in traditional medicine for its medicinal properties in the treatment and prevention of a variety of ailments. This study aims to extract and to separate the phytochemical compounds from the bark of P. granatum, to identify them and to study the inhibitory effect of its extracts against antidiabetic activity. First, we carried out successive hot extractions with solvents (chloroform, acetone, methanol, and water) of increasing polarity by the Soxhlet. Then, using both qualitative and quantitative phytochemical investigation, we were able to identify groups of chemicals that were present in all extracts. We identified the majority of the molecular structures of chemicals found in each extract using HPLC-DAD analysis. The inhibition against both intestinal α-glucosidase and pancreatic α-amylase enzymes by P. granatum extracts was used to evaluate their potential antidiabetic effect in vitro. Our results demonstrated the great potential of the acetone extract. Ellagic acid, (−)-catechin, vanillin and vanillic acid were proposed as the most active compounds by the correlation analysis, and their actions were confirmed through the calculation of their IC50 and the determination of their inhibition mechanisms by molecular modelling. To summarize, these results showed that P. granatum bark, a natural agro-industrial by-product, may constitute a promising option for antidiabetic therapeutic therapy.

Eclipta alba L., also known as false daisy, is well known and commercially attractive plant with excellent hepatotoxic and antidiabetic activities. Light is considered a key modulator in plant morphogenesis and survival by regulating important physiological cascades. Current study was carried out to investigate growth and developmental aspects of E. alba under differential effect of multispectral lights. In vitro derived callus culture of E. alba was exposed to multispectral monochromatic lights under controlled aseptic conditions. Maximum dry weight was recorded in culture grown under red light (11.2 g/L) whereas negative effect was observed under exposure of yellow light on callus growth (4.87 g/L). Furthermore, red light significantly enhanced phenolics and flavonoids content (TPC: 57.8 mg/g, TFC: 11.1 mg/g) in callus cultures compared to rest of lights. HPLC analysis further confirmed highest accumulation of four major compounds i.e. coumarin (1.26 mg/g), eclalbatin (5.00 mg/g), wedelolactone (32.54 mg/g) and demethylwedelolactone (23.67 mg/g) and two minor compounds (β-amyrin: 0.38 mg/g, luteolin: 0.39 mg/g) in red light treated culture whereas stigmasterol was found optimum (0.22 mg/g) under blue light. In vitro based biological activities including antioxidant, antidiabetic and lipase inhibitory assays showed optimum values in cultures exposed to red light, suggesting crucial role of these phytochemicals in the enhancement of the therapeutic potential of E. alba. These results clearly revealed that the use of multispectral lights in in vitro cultures could be an effective strategy for enhanced production of phytochemicals.

Nelumbo nucifera Gaertn., or the so-called sacred lotus, is a useful aquatic plant in the Nelumbonaceae family that has long been used to prepare teas, traditional medicines as well as foods. Many studies reported on the phytochemicals and biological activities of its leaves and seeds. However, to date, only few studies were conducted on its stamen, which is the most important ingredient for herbal medicines, teas and other phytopharmaceutical products. Thus, this present study focuses on the following: (1) the application of high-performance liquid chromatography with photodiode array detection for a validated separation and quantification of flavonoids from stamen; (2) the Nelumbo nucifera stamen’s in vitro and in cellulo antioxidant activities; as well as (3) its potential regarding the inhibition of skin aging enzymes for cosmetic applications. The optimal separation of the main flavonoids from the stamen ethanolic extract was effectively achieved using a core-shell column. The results indicated that stamen ethanolic extract has higher concentration of in vitro and in cellulo antioxidant flavonoids than other floral components. Stamen ethanolic extract showed the highest protective effect against reactive oxygen/nitrogen species formation, as confirmed by cellular antioxidant assay using a yeast model. The evaluation of potential skin anti-aging action showed that the stamen extract has higher potential to inhibit tyrosinase and collagenase compared with its whole flower. These current findings are the first report to suggest the possibility to employ N. nucifera stamen ethanolic extract as a tyrosinase and collagenase inhibitor in cosmetic applications, as well as the utility of the current separation method.

Medicinal plants are an inevitable source of pharmaceutical drugs and most of the world population depends on these plants for health benefits. The increasing global demand for bioactive compounds from medicinal plants has posed a great threat to their existence due to overexploitation. Adventitious root and hairy root culture systems are an alternative approach to the conventional method for mass production of valuable compounds from medicinal plants owing to their rapid growth, biosynthetic and genetic stability. The main purpose of this review is to investigate the recent scientific research published worldwide on the application of adventitious and hairy root cultures to produce valuable compounds from medicinal plants. Furthermore, a comparison of adventitious root vs. hairy root cultures to produce valuable compounds has also been discussed. Various aspects such as medium composition, carbon source, pH, amount of macronutrients, optimization strategy, scale-up cultures, and use of biotic abiotic and nano-elicitors at various concentrations are the topic of discussion in this review. Several studies on adventitious and hairy root cultures of Polygonum multiflorum¸ Withania somnifera¸ Echinacea purpurea and Ajuga bracteosa have been discussed in detail which highlights the importance of elicitation strategies and bioreactor system, presenting commercial applications.

Nelumbo nucifera is one of the most valuable medicinal species of the Nelumbonaceae family that has been consumed since the ancient historic period. Its stamen is an indispensable ingredient for many recipes of traditional medicines, and has been proved as a rich source of flavonoids that may provide an antiaging action for pharmaceutical or medicinal applications. However, there is no intense study on antiaging potential and molecular mechanisms. This present study was designed to fill in this important research gap by: (1) investigating the effects of sacred lotus stamen extract (LSE) on yeast lifespan extension; and (2) determining their effects on oxidative stress and metabolism to understand the potential antiaging action of its flavonoids. A validated ultrasound-assisted extraction method was also employed in this current work. The results confirmed that LSE is rich in flavonoids, and myricetin-3-O-glucose, quercetin-3-O-glucuronic acid, kaempferol-3-O-glucuronic acid, and isorhamnetin-3-O-glucose are the most abundant ones. In addition, LSE offers a high antioxidant capacity, as evidenced by different in vitro antioxidant assays. This present study also indicated that LSE delayed yeast (Saccharomyces cerevisiae, wild-type strain DBY746) chronological aging compared with untreated control yeast and a positive control (resveratrol) cells. Moreover, LSE acted on central metabolism, gene expressions (SIR2 and SOD2), and enzyme regulation (SIRT and SOD enzymatic activities). These findings are helpful to open the door for the pharmaceutical and medical sectors to employ this potential lotus raw material in their future pharmaceutical product development.

Asian lotus has long been consumed as a food and herbal drug that provides several health benefits. The number of studies on its biological activity is significant, but research at the population level to investigate the variation in phytochemicals and biological activity of each population which is useful for a more efficient phytopharmaceutical application strategy remains needed. This present study provided the frontier results to fill-in this necessary gap to investigating the phytopharmaceutical potential of perianth and stamen, which represent an important part for Asian traditional medicines, from 18 natural populations throughout Thailand by (1) determining their phytochemical profiles, such as total contents of phenolic, flavonoid, and anthocyanin, and (2) determining the antioxidant activity of these natural populations using various antioxidant assays to examine different mechanisms. The result showed that Central is the most abundant floristic region. The stamen was higher in total phenolic and flavonoid contents, whereas perianth was higher in monomeric anthocyanin content. This study provided the first description of the significant correlation between phytochemical contents in perianth compared with stamen extracts, and indicated that flavonoids are the main phytochemical class. This analysis indicated that the stamen is a richer source of flavonoids than perianth, and provided the first report to quantify different flavonoids accumulated in stamen and perianth extracts under their native glycosidic forms at the population level. Various antioxidant assays revealed that major flavonoids from N. nucifera prefer the hydrogen atom transfer mechanism when quenching free radicals. The significant correlations between various phytochemical classes and the different antioxidant tests were noted by Pearson correlation coefficients and emphasized that the antioxidant capability of an extract is generally the result of complex phytochemical combinations as opposed to a single molecule. These current findings offer the alternative starting materials to assess the phytochemical diversity and antioxidant potential of N. nucifera for phytopharmaceutical sectors.

Fabaceae is the third largest family containing great variation among populations. However, previous studies mainly focus on single species, and phytochemicals at population level have never been reported. This work aims to complete this knowledge with 50 populations from throughout Thailand by (1) determining total phenolic (TPC), flavonoid (TFC), and anthocyanin (TAC) contents; and (2) investigating in vitro and cellular antioxidant potentials. Phytochemicals of 50 populations from different localities are differed, illustrating high heterogeneity occurring in polyphenols accumulations. Vigna unguiculata subsp. sesquipedalis populations showed low variability in TPC ranging from 628.3 to 717.3 mg/100 g DW gallic acid equivalent, whereas the high variability found in TFC and TAC range from 786.9 to 1536.1 mg/100 g DW quercetin equivalent, and 13.4 to 41.6 mg/100 g DW cyanidin equivalent. Red cultivar population #16 had the greatest TAC, but surprisingly the cream cultivars were relatively high in anthocyanins. HPLC quantification of genistein and daidzein showed great variations among populations. In vitro antioxidant results indicated that antioxidant capacity mediated by electron transfer. Cellular antioxidants ranged from 59.7% to 87.9% of ROS/RNS in yeast model. This study investigated at the population level contributing to better and frontier knowledge for nutraceutical/phytopharmaceutical sectors to seek potential raw plant material.

Pigeon pea is an important pea species in the Fabaceae family that has long been used for food, cosmetic, and other phytopharmaceutical applications. Its seed is reported as a rich source of antioxidants and anti-inflammatory flavonoids, especially isoflavones, i.e., cajanin, cajanol, daidzein, and genistein. In today’s era of green chemistry and green cosmetic development, the development and optimization of extraction techniques is increasing employed by the industrial sectors to provide environmentally friendly products for their customers. Surprisingly, there is no research report on improving the extraction of these isoflavonoids from pigeon pea seeds. In this present study, ultrasound-assisted extraction (USAE) methodology, which is a green extraction that provides a shorter extraction time and consumes less solvent, was optimized and compared with the conventional methods. The multivariate strategy, the Behnken–Box design (BBD) combined with response surface methodology, was employed to determine the best extraction conditions for this USAE utilizing ethanol as green solvent. Not only in vitro but also cellular antioxidant activities were evaluated using different assays and approaches. The results indicated that USAE provided a substantial gain of ca 70% in the (iso)flavonoids extracted and the biological antioxidant activities were preserved, compared to the conventional method. The best extraction conditions were 39.19 min with a frequency of 29.96 kHz and 63.81% (v/v) aqueous ethanol. Both the antioxidant and anti-aging potentials of the extract were obtained under optimal USAE at a cellular level using yeast as a model, resulting in lower levels of malondialdehyde. These results demonstrated that the extract can act as an effective activator of the cell longevity protein (SIR2/SIRT1) and cell membrane protector against oxidative stress. This finding supports the potential of pigeon pea seeds and USAE methodology to gain potential antioxidant and anti-aging (iso)flavonoids-rich sources for the cosmetic and phytopharmaceutical sectors.

The edible beans in Fabaceae have been used for foods and medicines since the ancient time, and being used more and more. It is also appeared as a major ingredient in dairy cooking menu in many regions including Thailand, a rich biodiversity country. Many studies reported on health benefits of their flavonoids, but there is no report on the effect of cooking on phytochemical profile and pharmacological potentials. Thus, this present study aims to complete this knowledge, with the 10 most consumed Fabaceae beans in Thailand, by determining the impact of traditional cooking and gastrointestinal digestion on their phytochemicals, their antioxidant and anti-diabetic activities using different in vitro and in cellulo yeast models. The results showed that Vigna unguiculata subsp. sesquipedalis were the richest source of phytochemicals, whereas the population of V. mungo, Phaseolus vulgaris, V. angularis, and V. unguiculata subsp. sesquipedalis were richest in monomeric anthocyanin contents (MAC). Furthermore, the results clearly demonstrated the impact of the plant matrix effect on the preservation of a specific class of phytochemicals. In particular, after cooking and in vitro digestion, total flavonoid contents (TFC) in Glycine max extract was higher than in the uncooked sample. This study is the first report on the influence of cooking and in vitro gastrointestinal digestion on the inhibition capacity toward advanced glycation end products (AGEs). All samples showed a significant capacity to stimulate glucose uptake in yeast model, and V. angularis showed the highest capacity. Interestingly, the increase in glucose uptake after in vitro digestion was higher than in uncooked samples for both P. vulgaris and G. max samples. The current study is the first attempt to investigate at the effects of both processes not only on the natural bioactive compounds but also on antioxidant and anti-diabetic activities of Thailand’s 10 most consumed beans that can be applied for agro-industrial and phytopharmaceutical sectors.

The green synthesis of nanoparticles has emerged as a simple, safe, sustainable, reliable and eco-friendly protocol. Among different types of NPs, green-synthesized zinc oxide NPs (ZnONPs) show various promising biological uses due to their interesting magnetic, electrical, optical and chemical characteristics. Keeping in view the dependence of the therapeutic efficacy of NPs on their physico-chemical characteristics, the green synthesis of ZnONPs using Casuarina equisetifolia leaf extract under UV-A and UV-C light was carried out in this study. UV-irradiation helped to control the size and morphology of ZnONPs by exciting the electrons in the photoactive compounds of plant extracts to enhance the bio-reduction of ZnO into ZnONPs. C. equisetifolia leaf extract was found enriched with phenolic (2.47 ± 0.12 mg GAE/g DW) and flavonoid content (0.88 ± 0.28 mg QE/g DW) contributing to its 74.33% free-radical scavenging activity. FTIR spectra showed the involvement of polyphenols in the bio-reduction, stabilization and capping of ZnONPs. Moreover, SEM-EDX and XRD analyses showed great potential of UV-C light in yielding smaller (34–39 nm) oval-shaped ZnONPs, whereas UV-A irradiation resulted in the formation of fairly spherical 67–71 nm ZnONPs and control ZnONPs were of mixed shape and even larger size (84–89 nm). Green-synthesized ZnONPs, notably CE-UV-C-ZnONPs, showed promising anti-bacterial activities against Bacillus subtilis, Pseudomonas fluorescens and Pseudomonas aeruginosa. Moreover, ZnONPs also enhanced ROS production which led to a significant loss of mitochondrial membrane potential and activated caspase-3 gene expression and caspase-3/7 activity in human hepatocellular carcinoma (HepG2) cells. CE-UV-C-ZnONP treatment reduced HepG2 cell viability to as low as 36.97% owing to their unique shape and smaller size. Lastly, ZnONPs were found to be highly biocompatible towards brine shrimp and human red blood cells suggesting their bio-safe nature. This research study sheds light on the plausible role of UV radiation in the green synthesis of ZnONPs with reasonable control over their size and morphology, thus improving their biological efficacy.

Fagonia indica is a rich source of pharmacologically active compounds. The variation in the metabolites of interest is one of the major issues in wild plants due to different environmental factors. The addition of chemical elicitors is one of the effective strategies to trigger the biosynthetic pathways for the release of a higher quantity of bioactive compounds. Therefore, this study was designed to investigate the effects of chemical elicitors, aluminum chloride (AlCl3) and cadmium chloride (CdCl2), on the biosynthesis of secondary metabolites, biomass, and the antioxidant system in callus cultures of F. indica. Among various treatments applied, AlCl3 (0.1 mM concentration) improved the highest in biomass accumulation (fresh weight (FW): 404.72 g/L) as compared to the control (FW: 269.85 g/L). The exposure of cultures to AlCl3 (0.01 mM) enhanced the accumulation of secondary metabolites, and the total phenolic contents (TPCs: 7.74 mg/g DW) and total flavonoid contents (TFCs: 1.07 mg/g DW) were higher than those of cultures exposed to CdCl2 (0.01 mM) with content levels (TPC: 5.60 and TFC: 0.97 mg/g) as compared to the control (TPC: 4.16 and TFC: 0.42 mg/g DW). Likewise, AlCl3 and CdCl2 also promoted the free radical scavenging activity (FRSA; 89.4% and 90%, respectively) at a concentration of 0.01 mM, as compared to the control (65.48%). For instance, the quantification of metabolites via high-performance liquid chromatography (HPLC) revealed an optimum production of myricetin (1.20 mg/g), apigenin (0.83 mg/g), isorhamnetin (0.70 mg/g), and kaempferol (0.64 mg/g). Cultures grown in the presence of AlCl3 triggered higher quantities of secondary metabolites than those grown in the presence of CdCl2 (0.79, 0.74, 0.57, and 0.67 mg/g). Moreover, AlCl3 at 0.1 mM enhanced the biosynthesis of superoxide dismutase (SOD: 0.08 nM/min/mg-FW) and peroxidase enzymes (POD: 2.37 nM/min/mg-FW), while CdCl2 resulted in an SOD activity up to 0.06 nM/min/mg-FW and POD: 2.72 nM/min/mg-FW. From these results, it is clear that AlCl3 is a better elicitor in terms of a higher and uniform productivity of biomass, secondary cell products, and antioxidant enzymes compared to CdCl2 and the control. It is possible to scale the current strategy to a bioreactor for a higher productivity of metabolites of interest for various pharmaceutical industries.

Due to the high volume of peel produced, Citrus by-product processing could be a significant source of phenolic compounds, in addition to essential oil. Citrus fruit residues, which are usually dumped as waste in the environment, could be used as a source of nutraceuticals. Citrus aurantium (L), also known as sour or bitter orange, is a member of the Rutaceae family and is the result of interspecific hybridization between Citrus reticulata and Citrus maxima. The purpose of this study is to chemically and biologically evaluate the peel of C. aurantium, which is considered a solid waste destined for abandonment. To achieve more complete extraction of the phytochemicals, we used a sequential extraction process with Soxhlet using the increasing polarity of solvents (i.e., cyclohexane, chloroform, ethyl acetate, acetone, and ethanol–water mixture). Essential oil (EO) from the Citrus peel, which was present at 1.12%, was also prepared by hydrodistillation for comparison. Various phytochemical assays were used to determine the qualitative chemical composition, which was subsequently characterized using GC-MS and HPLC-DAD. The inhibitory effects of C. aurantium peel extract on two enzymes, intestinal α-glucosidase and pancreatic α-amylase, were measured in vitro to determine their potential hypoglycemic and antidiabetic actions. Each extract had a significantly different phytochemical composition. According to GC-MS analyses, which allow the identification of 19 compounds, d-limonene is the most abundant compound in both EO and cyclohexane extract, at 35.17% and 36.15% (w/w). This comparison with hydrodistillation shows the value of the sequential process in extracting this valuable terpene in large quantities while also allowing for the subsequent extraction of other bioactive substances. On the contrary, linoleic acid is abundant (54.35% (w/w)) in ethyl acetate extract (EAE) with a lower amount of d-limonene. HPLC-DAD analysis allows the identification of 11 phytochemicals, with naringenin being the most abundant flavanone, detected in acetone extract (ACE) (23.94% (w/w)), ethanol–water extract mixture (EWE) (28.71% (w/w)), and chloroform extract (CFE) (30.20% (w/w)). Several extracts significantly inhibited α-amylase and/or α-glycosidase in vitro. At a dose of 332 g/mL, ACE, CFE, and EWE inhibited the two enzymes by approximately 98%. There were strong significant correlations between naringenin and α-glucosidase inhibition and between gallic acid and α-amylase inhibition. Molecular docking experiments further verified this. Finally, oral administration of C. aurantium extracts at a dose of 2000 mg/kg did not cause any effect on mice mortality or signs of acute toxicity, indicating that it is non-toxic at these doses. These findings suggest that C. aurantium peels could be a valuable by-product by providing a rich source of non-toxic phytoconstituents, particularly those with potential antidiabetic action that needs to be confirmed in vivo.

Nigella sativa L. (Ranunculaceae), commonly referred to as black seeds or black cumin, is used in popular medicine (herbal) all over the world for the treatment and prevention of several diseases, including diabetes. This study aims to investigate the inhibitory effect of N. sativa extracts and fractions against the activities of intestinal α-glucosidase and pancreatic α-amylase in vitro, and to explain the inhibitory effect of these fractions against these enzymes by identifying their active compounds responsible for this effect and determine their modes of inhibition. To do so, N. sativa hexane and acetone extracts were prepared and analyzed by GC–MS and HPLC–DAD, respectively. The hexane extract was further fractioned into eight different fractions, while the acetone extract generated eleven fractions. The extracts as well as the resulting fractions were characterized and evaluated for their potential in vitro antidiabetic activity using intestinal α-glucosidase and pancreatic α-amylase inhibitory assays in vitro. Hexane extract and fractions were less active than acetone extract and fractions. In the case of intestinal α-glucosidase activity, the acetone fraction SA3 had a high inhibitory effect on intestinal α-glucosidase activity with 72.26 ± 1.42%, comparable to the effect of acarbose (70.90 ± 1.12%). For the pancreatic α-amylase enzymatic inhibitory assay, the acetone fractions showed an inhibitory capacity close to that for acarbose. In particular, the SA2 fraction had an inhibitory effect of 67.70 ± 0.58% and was rich in apigenin and gallic acid. From these fractions, apigenin, (−)-catechin, and gallic acid were further characterized for their inhibitory actions. IC50 and inhibition mode were determined by analyzing enzyme kinetic parameters and by molecular modeling. Interestingly, (−)-catechin showed a possible synergistic effect with acarbose toward α-glucosidase enzyme inhibition, whereas apigenin showed an additive effect with acarbose toward α-amylase enzymatic inhibition. Furthermore, we studied the toxicity of N. sativa hexane and acetone extracts as well as that of acetone fractions. The result of acute toxicity evaluation demonstrated that N. sativa extracts were nontoxic up to a concentration of 10 g/kg, except for fraction SA3. Taken together, these results indicate that N. sativa extracts and/or derived compounds could constitute promising nutraceuticals for the prevention and treatment of type 2 diabetes mellitus.

In vitro cultures of scarlet flax (Linum grandiflorum L.), an important ornamental flax, have been established as a new possible valuable resource of lignans and neolignans for antioxidant and anti-inflammatory applications. The callogenic potential at different concentrations of α-naphthalene acetic acid (NAA) and thidiazuron (TDZ), alone or in combinations, was evaluated using both L. grandiflorum hypocotyl and cotyledon explants. A higher callus induction frequency was observed on NAA than TDZ, especially for hypocotyl explants, with a maximum frequency (i.e., 95.2%) on 1.0 mg/L of NAA. The presence of NAA (1.0 mg/L) in conjunction with TDZ tended to increase the frequency of callogenesis relative to TDZ alone, but never reached the values observed with NAA alone, thereby indicating the lack of synergy between these two plant growth regulators (PGRs). Similarly, in terms of biomass, NAA was more effective than TDZ, with a maximum accumulation of biomass registered for medium supplemented with 1.0 mg/L of NAA using hypocotyls as initial explants (DW: 13.1 g). However, for biomass, a synergy between the two PGRs was observed, particularly for cotyledon-derived explants and for the lowest concentrations of TDZ. The influence of these two PGRs on callogenesis and biomass is discussed. The HPLC analysis confirmed the presence of lignans (secoisolariciresinol (SECO) and lariciresinol (LARI) and neolignan (dehydrodiconiferyl alcohol [DCA]) naturally accumulated in their glycoside forms. Furthermore, the antioxidant activities performed for both hypocotyl- and cotyledon-derived cultures were also found maximal (DPPH: 89.5%, FRAP 866: µM TEAC, ABTS: 456 µM TEAC) in hypocotyl-derived callus cultures as compared with callus obtained from cotyledon explants. Moreover, the anti-inflammatory activities revealed high inhibition (COX-1: 47.4% and COX-2: 51.1%) for extract of hypocotyl-derived callus cultures at 2.5 mg/L TDZ. The anti-inflammatory action against COX-1 and COX-2 was supported by the IC50 values. This report provides a viable approach for enhanced biomass accumulation and efficient production of (neo)lignans in L. grandiflorum callus cultures.

Plant secondary metabolites are known to have a variety of biological activities beneficial to human health. They are becoming more popular as a result of their unique features and account for a major portion of the pharmacological industry. However, obtaining secondary metabolites directly from wild plants has substantial drawbacks, such as taking a long time, posing a risk of species extinction owing to over-exploitation, and producing a limited quantity. Thus, there is a paradigm shift towards the employment of plant tissue culture techniques for the production of key secondary metabolites in vitro. Elicitation appears to be a viable method for increasing phytochemical content and improving the quality of medicinal plants and fruits and vegetables. In vitro culture elicitation activates the plant’s defense response and increases the synthesis of secondary metabolites in larger proportions, which are helpful for therapeutic purposes. In this respect, light has emerged as a unique and efficient elicitor for enhancing the in vitro production of pharmacologically important secondary metabolites. Various types of light (UV, fluorescent, and LEDs) have been found as elicitors of secondary metabolites, which are described in this review.

Silybum marianum (L.) Gaertn is a rich source of antioxidants and anti-inflammatory flavonolignans with great potential for use in pharmaceutical and cosmetic products. Its biotechnological production using in vitro culture system has been proposed. Chitosan is a well-known elicitor that strongly affects both secondary metabolites and biomass production by plants. The effect of chitosan on S. marianum cell suspension is not known yet. In the present study, suspension cultures of S. marianum were exploited for their in vitro potential to produce bioactive flavonolignans in the presence of chitosan. Established cell suspension cultures were maintained on the same hormonal media supplemented with 0.5 mg/L BAP (6-benzylaminopurine) and 1.0 mg/L NAA (α-naphthalene acetic acid) under photoperiod 16/8 h (light/dark) and exposed to various treatments of chitosan (ranging from 0.5 to 50.0 mg/L). The highest biomass production was observed for cell suspension treated with 5.0 mg/L chitosan, resulting in 123.3 ± 1.7 g/L fresh weight (FW) and 17.7 ± 0.5 g/L dry weight (DW) productions. All chitosan treatments resulted in an overall increase in the accumulation of total flavonoids (5.0 ± 0.1 mg/g DW for 5.0 mg/L chitosan), total phenolic compounds (11.0 ± 0.2 mg/g DW for 0.5 mg/L chitosan) and silymarin (9.9 ± 0.5 mg/g DW for 0.5 mg/L chitosan). In particular, higher accumulation levels of silybin B (6.3 ± 0.2 mg/g DW), silybin A (1.2 ± 0.1 mg/g DW) and silydianin (1.0 ± 0.0 mg/g DW) were recorded for 0.5 mg/L chitosan. The corresponding extracts displayed enhanced antioxidant and anti-inflammatory capacities: in particular, high ABTS antioxidant activity (741.5 ± 4.4 μM Trolox C equivalent antioxidant capacity) was recorded in extracts obtained in presence of 0.5 mg/L of chitosan, whereas highest inhibitions of cyclooxygenase 2 (COX-2, 30.5 ± 1.3 %), secretory phospholipase A2 (sPLA2, 33.9 ± 1.3 %) and 15-lipoxygenase (15-LOX-2, 31.6 ± 1.2 %) enzymes involved in inflammation process were measured in extracts obtained in the presence of 5.0 mg/L of chitosan. Taken together, these results highlight the high potential of the chitosan elicitation in the S. marianum cell suspension for enhanced production of antioxidant and anti-inflammatory silymarin-rich extracts.

With the increase in biotechnological, environmental, and nutraceutical importance of algae, about 100 whole genomic sequences of algae have been published, and this figure is expected to double in the coming years. The phenotypic and ecological diversity among algae hints at the range of functional capabilities encoded by algal genomes. In order to explore the biodiversity of algae and fully exploit their commercial potential, understanding their evolutionary, structural, functional, and developmental aspects at genomic level is a pre-requisite. So forth, the algal genomic analysis revealed us that algae evolved through endosymbiotic gene transfer, giving rise to around eight phyla. Amongst the diverse algal species, the unicellular green algae Chlamydomonas reinhardtii has attained the status of model organism as it is an ideal organism to elucidate the biological processes critical to plants and animals, as well as commercialized to produce range of bio-products. For this review, an overview of evolutionary process of algae through endosymbiosis in the light of genomics, as well as the phylogenomic, studies supporting the evolutionary process of algae was reviewed. Algal genomics not only helped us to understand the evolutionary history of algae but also may have an impact on our future by helping to create algae-based products and future biotechnological approaches.

Thai basil is a renowned medicinal plant and a rich source of bioactive antioxidant compounds with several health benefits, with actions to prevent of cancer, diabetes and cardiovascular disease. Plant cell and tissue culture technologies can be routinely established as an important, sustainable and low-cost biomass source to produce high-value phytochemicals. The current study aimed at developing an effective protocol to produce Thai basil leaf-derived callus cultures with sustainable and high production of biomass and antioxidants as an alternative of leaves production. MS basal medium with various concentrations of plant growth regulators (PGRs) compatible with nutraceutical applications (i.e., gibberellic acid (GA3) and 6-benzylaminopurine (BAP) either alone or in combination with naphthalene acetic acid (NAA)) were evaluated. Among all tested PGRs, the combination BAP:NAA (5 mg/L:1 mg/L) yields the maximum biomass accumulation (fresh weight (FW): 190 g/L and dry weight (DW): 13.05 g/L) as well as enhanced phenolic (346.08 mg/L) production. HPLC quantification analysis indicated high productions of chicoric acid (35.77 mg/g DW) and rosmarinic acid (7.35 mg/g DW) under optimized callus culture conditions. Antioxidant potential was assessed using both in vitro cell free and in vivo cellular antioxidant assays. Maximum in vitro antioxidant activity DPPH (93.2% of radical scavenging activity) and ABTS (1322 µM Trolox equivalent antioxidant capacity) was also observed for the extracts from callus cultures grown in optimal conditions. In vivo cellular antioxidant activity assay confirmed the effective protection against oxidative stress of the corresponding extract by the maximum inhibition of ROS and RNS production. Compared to commercial leaves, callus extracts showed higher production of chicoric acid and rosmarinic acid associated with higher antioxidant capacity. In addition, this biological system also has a large capacity for continuous biomass production, thus demonstrating its high potential for possible nutraceutical applications.

“Bau Luang” or Nelumbo nucifera Gaertn. is an aquatic medicinal herb that has been used as a component of traditional medicines, medicinal products, and herbal tea for good health, particularly in Asia. The stamen of N. nucifera is an important part of this medicinal plant that is used in the form of dried and/or powdered stamens for herbal tea as well as the main ingredient of some traditional remedies. However, there is another aquatic herb called “Bau Sai” or Nymphaea lotus L. that is distributed in similar locations. Living plants of these two aquatic species may be classified according to their morphology, but the dried and powdered stamens of these two medicinal species are difficult to distinguish. The major reason of adulteration is the higher price of Bau Luang stamen. As a result, various methods of authentication, such as pollen micromorphology evaluation using scanning electron microscopy (SEM) analysis, bioinformatics analysis of two nuclear and plastic DNA markers, phytochemical stamen profiling, and Fourier transform infrared (FTIR) analysis of stamen plant material authentication from Bau Luang and Bau Sai, have been used in this present research in order to avoid some adulteration and/or misuse between the dried stamens of Bau Luang and Bau Sai. These results showed that the micro-morphology of pollen (size of pollen grain, number of apertures, and surface ornamentation) from the SEM analysis, some phytochemical compounds and the FTIR sporopollenin-to-protein ratio signal analysis are potential tools for authentication and identification of these two medicinal plants from their dried-stamen materials. This model of investigation may also be used to distinguish dried plant material from other problematic plant groups.

Nymphaea lotus L. is the medicinal plant that has long been used for food, cosmetics and traditional medicines in Africa and Asia since ancient times. Its flavonoids and other interesting phytochemical compounds from rhizome, leaf and the whole flowers have been reported in the previous published research. However, stamens, which are essential for reproductive functions, may also represent new alternative sources of potential antioxidant flavonoids, as investigated in this study. The innovative green chemistry methods, i.e., ultrasound-assisted extraction (UAE) as well as a macroporous resin (MPR) purification procedure, were employed in this current research. Using a full factorial design coupled to three-dimensional (3D) surface plot methodology, the influence of three variables, namely aqEtOH concentration (ranging from 50 to 100% (v/v), US frequency (ranging from 0 (no US applied) to 45 kHz), and the extraction duration (ranging from 20 to 60 min), were evaluated. Five MPRs with different surface areas, average pore diameters, matrix types and polarities were also investigated for the purification of total flavonoids. The optimal UAE condition is 90% (v/v) aqEtOH with 34.65 khz ultrasonic frequency and 46 min of extraction duration. Compared with the conventional heat reflux extraction (HRE) method, a significant 1.35-fold increase in total flavonoids content was obtained using optimized UAE conditions (169.64 for HRE vs. 235.45 mg/g dry weight for UAE), causing a 2.80-fold increase when this UAE associated with MPR purification (475.42 mg/g dry weight). In vitro cell free antioxidant activity of N. lotus stamen extracts and in cellulo antioxidant investigation using yeast model showed the same trend, indicating that the best antioxidant flavonoid can be found in UAE coupled with MPR purification. Moreover, in the yeast model, the expression of key antioxidant genes such as SIR2 and SOD2 were expressed at the highest level in yeast cells treated with the extract from UAE together with MPR purification. Consequently, it can be seen that the UAE combined with MPR purification can help enhance the flavonoid antioxidant potential of the stamens extract from this medicinal species.

Linum usitatissimum biosynthesizes lignans and neolignans that are diet and medicinally valuable metabolites. In recent years, zinc oxide nanoparticles (ZnONPs) have emerged as potential elicitors for the enhanced biosynthesis of commercial secondary metabolites. Herein, we investigated the influence of biogenic ZnONPs on both seedlings and stem-derived callus of L. usitatissimum. Seedlings of L. usitatissimum grown on Murashige and Skoog (MS) medium supplemented with ZnONPs (1–1000 mg/L) presented the highest antioxidant activity, total phenolic content, total flavonoid content, peroxidase and superoxide dismutase activities at 500 mg/L, while the maximum plantlet length was achieved with 10 mg/L. Likewise, the high-performance liquid chromatography (HPLC) analysis revealed the enhanced production of secoisolariciresinol diglucoside, lariciresinol diglucoside, dehydrodiconiferyl alcohol glucoside and guaiacylglycerol-β-coniferyl alcohol ether glucoside in the plantlets grown on the 500 mg/L ZnONPs. On the other hand, the stem explants were cultured on MS media comprising 1-naphthaleneacetic acid (1 mg/L) and ZnONPs (1–50 mg/L). The highest antioxidant and other activities with an enhanced rooting effect were noted in 25 mg/L ZnONP-treated callus. Similarly, the maximum metabolites were also accumulated in 25 mg/L ZnONP-treated callus. In both systems, the dose-dependent production of reactive oxygen species (ROS) was recorded, resulting in oxidative damage with a more pronounced toxic effect on in vitro cultures. Altogether, the results from this study constitute a first comprehensive view of the impact of ZnONPs on the oxidative stress and antioxidant responses in seedlings vs. in vitro cultures.

Solanum xanthocarpum is considered an important traditional medicinal herb because of its unique antioxidant, and anti-diabetic, anti-aging, and anti-inflammatory potential. Because of the over exploitation linked to its medicinal properties as well as destruction of its natural habitat, S. xanthocarpum is now becoming endangered and its supply is limited. Plant in vitro culture and elicitation are attractive alternative strategies to produce biomass and stimulate biosynthesis of medicinally important phytochemicals. Here, we investigated the potential influence of seven different monochromatic light treatments on biomass and secondary metabolites accumulation in callus culture of S. xanthocarpum as well as associated biological activities of the corresponding extracts. Among different light treatments, highest biomass accumulation was observed in white light-treated callus culture. Optimum accumulation of total flavonoid contents (TFC) and total phenolic contents (TPC) were observed in callus culture kept under continuous white and blue light respectively than control. Quantification of phytochemicals through HPLC revealed that optimum production of caffeic acid (0.57 ± 0.06 mg/g DW), methyl-caffeate (17.19 mg/g ± 1.79 DW), scopoletin (2.28 ± 0.13 mg/g DW), and esculetin (0.68 ± 0.07 mg/g DW) was observed under blue light callus cultures. Compared to the classic photoperiod condition, caffeic acid, methyl-caffeate, scopoletin, and esculetin were accumulated 1.7, 2.5, 1.1, and 1.09-folds higher, respectively. Moreover, high in vitro cell free antioxidant, anti-diabetic, anti-aging, and anti-inflammatory activities were closely associated with the production of these secondary metabolites. These results clearly showed the interest to apply multispectral light as elicitor of in vitro callus cultures S. xanthocarpum to promote the production of important phytochemicals, and allow us to propose this system as an alternative for the collection of this endangered species from the wild.

Grape canes are waste biomass of viticulture containing bioactive polyphenols valuable in cosmetics. Whereas several studies reported the cosmetic activities of E-resveratrol, only few described the potential of E-ε-viniferin, the second major constituent of grape cane extracts (GCE), and none of them investigated GCE as a natural blend of polyphenols for cosmetic applications. In this study, we considered the potential of GCE from polyphenol-rich grape varieties as multifunctional cosmetic ingredients. HPLC analysis was performed to quantify major polyphenols in GCE i.e., catechin, epicatechin, E-resveratrol, E-piceatannol, ampelopsin A, E-ε-viniferin, hopeaphenol, isohopeaphenol, E-miyabenol C and E-vitisin B from selected cultivars. Skin whitening potential through tyrosinase inhibition assay and the activation capacity of cell longevity protein (SIRT1) of GCE were compared to pure E-resveratrol and E-ε-viniferin. Drug-likeness of GCE polyphenols were calculated, allowing the prediction of skin permeability and bioavailability. Finally, the present data enabled the consideration of GCE from polyphenol-rich varieties as multifunctional cosmetic ingredients in accordance with green chemistry practices.

Almond (Prunus dulcis (Mill.) D.A. Webb) is one of the most important nut crops both in terms of area and production. Over the last few decades, an important part of the beneficial actions for health associated with their consumption was attributed to the phenolic compounds, mainly accumulated in almond skin. Interestingly, after cold-pressed oil extraction, most of these antioxidant phenolic compounds are retained in a skin-enriched by-product, a so-called almond cold-pressed oil residue. In Morocco, the fifth highest ranking producer in the world, this production generates an important part of this valuable byproduct. In the present study, using a multivariate Box–Behnken design, an ultrasound-assisted extraction method of phenolic compounds from Moroccan almond cold-pressed oil residue was developed and validated. Response surface methodology resulted in the optimal extraction conditions: the use of aqueous ethanol 53.0% (v/v) as a green solvent, applying an ultrasound frequency of 27.0 kHz for an extraction duration of 29.4 min. The present ultrasound-assisted extraction allowed substantial gains in terms of extraction efficiency compared to conventional heat reflux extraction. Applied to three different local Beldi genotypes growing at three different experimental sites, the optimal conditions for ultrasound-assisted extraction led to a total phenolic content of 13.86 mg/g dry weight. HPLC analysis revealed that the main phenolic compounds from this valuable byproduct were: chlorogenic acid followed by protocatechuic acid, p-hydroxybenzoic acid, and p-coumaric acid. The accumulation of these phenolic compounds appeared to be more dependent on the genetic background than on the environmental impact here represented by the three experimental culture sites. Both in vitro cell free and cellular antioxidant assays were performed, and revealed the great potential of these extracts. In particular, correlation analysis provided evidence of the prominent roles of chlorogenic acid, protocatechuic acid, and p-hydroxybenzoic acid. To summarize, the validated ultrasound-assisted extraction method presented here is a quick, green, simple and efficient for the possible valorization of antioxidant phenolic compounds from Moroccan almond cold-pressed oil residues, making it possible to generate extracts with attractive antioxidant activities for future nutraceutical and/or cosmetic applications.

Almond (Prunus dulcis (Mill.) D.A.Webb) is one of the largest nut crops in the world. Recently, phenolic compounds, mostly stored in almond skin, have been associated with much of the health-promoting behavior associated with their intake. The almond skin enriched fraction obtained from cold-pressed oil residues of the endemic Moroccan Beldi ecotypes is particularly rich in chlorogenic acid. In this study, both almond skin extract (AE) and chlorogenic acid (CHL) supplements, similar to traditional positive control resveratrol, significantly increased the chronological life-span of yeast compared to the untreated group. Our results showed that AE and CHL significantly reduced the production of reactive oxygen and nitrogen species (ROS/RNS), most likely due to their ability to maintain mitochondrial function during aging, as indicated by the maintenance of normal mitochondrial membrane potential in treated groups. This may be associated with the observed activation of the anti-oxidative stress response in treated yeast, which results in activation at both gene expression and enzymatic activity levels for SOD2 and SIR2, the latter being an upstream inducer of SOD2 expression. Interestingly, the differential gene expression induction of mitochondrial SOD2 gene at the expense of the cytosolic SOD1 gene confirms the key role of mitochondrial function in this regulation. Furthermore, AE and CHL have contributed to the survival of yeast under UV-C-induced oxidative stress, by reducing the development of ROS/RNS, resulting in a significant reduction in cellular oxidative damage, as evidenced by decreased membrane lipid peroxidation, protein carbonyl content and 8-oxo-guanine formation in DNA. Together, these results demonstrate the interest of AE and CHL as new regulators in the chronological life-span and control of the oxidative stress response of yeast.

Flax lignans and neolignans impart health benefits, particularly in treating different types of cancers, due to their strong phytoestrogenic and antioxidant properties. The present study enhances the comprehension on the biosynthesis of antioxidant lignans and neolignans in root-derived in vitro cultures of flax (both callus and adventitious root). The results presented here clearly showed that the adventitious root culture efficiently produced a higher amount of lignans (at day 40) and neolignans (at day 30) than callus culture of flax. High performance liquid chromatography (HPLC) analysis revealed that the accumulations of secoisolariciresinol diglucoside (SDG, 5.5 mg g−1 DW (dry weight)) and dehydrodiconiferyl alcohol glucoside (DCG, 21.6 mg/g DW) were 2-fold higher, while guaiacylglycerol-β-coniferyl alcohol ether glucoside (GGCG, 4.9 mg/g DW) and lariciresinol glucoside (LDG, 11.9 mg/g DW) contents were 1.5-fold higher in adventitious root culture than in callus culture. Furthermore, the highest level of total phenolic production (119.01 mg/L), with an antioxidant free radical scavenging activity of 91.01%, was found in adventitious root culture at day 40, while the maximum level of total flavonoid production (45.51 mg/L) was observed in callus culture at day 30 of growth dynamics. These results suggest that adventitious root culture can be a good candidate for scaling up to industrial level to commercially produce these pharmacologically and nutritionally valuable metabolites.

The present study evaluated the interactive effect of melatonin and UV-C on phenylpropanoid metabolites profile and antioxidant potential of Ocimum basilicum L. Callus was treated with varying concentrations of melatonin and UV-C radiations for different time durations, either alone and/or in combination. Individual treatments of both UV-C and melatonin proved to be more effective than combine treatments. Results indicated that UV-C (10 min) exposure increased rosmarinic acid (134.5 mg/g dry weight (DW)), which was 2.3-fold greater than control. Chichoric acid (51.52 mg/g DW) and anthocyanin (cyanide 0.50 mg/g DW) were almost 4.1-fold, while peonidin was found 2.7-fold higher in UV-C (50 min) exposure. In the case of melatonin, 1.0 mg/L concentrations showed maximum rosmarinic acid (79.4 mg/g DW) accumulation; i.e., 1.4-fold more, as compared to the control. However, 2 mg/L melatonin accumulate chichoric acid (39.99 mg/g DW) and anthocyanin (cyanide: 0.45 mg/g DW and peonidin: 0.22 mg/g DW); i.e., 3.2, 3.7 and 2.0-fold increase, as compared to the control, respectively. On the other hand, melatonin-combined treatment (melatonin (Mel) (4 mg/L) + UV-C (20 min)) was proved to be effective in caffeic acid elicitation, which was 1.9-fold greater than the control. Furthermore, antioxidant potential was evaluated by both in vitro (DPPH, ABTS and FRAP assays) and in cellulo methods. Maximum in vitro antioxidant activity (DPPH: 90.6% and ABTS: 1909.5 µM) was observed for UV-C (50 min)-treated cultures. The highest in vitro antioxidant activity measured with the ABTS assay as compared to the FRAP assay, suggesting the main contribution of antioxidants from basil callus extracts acting through a hydrogen atom transfer (HAT) over an electron transfer (ET)-based mechanism. Cellular antioxidant assay was evaluated by production of ROS/RNS species using yeast cell cultures and further confirmed the protective action of the corresponding callus extracts against oxidative stress. Overall, both melatonin and UV-C are here proved to be effective elicitors since a positive correlation between the induced production of phenolic compounds, and in cellulo antioxidant action of basil callus extracts were observed.

Silybum marianum (L.) Gaertn. (aka milk thistle) constitutes the source of silymarin (SILM), a mixture of different flavonolignans and represents a unique model for their extraction. Here we report on the development and validation of an ultrasound-assisted extraction (UAE) method of S. marianum flavonolignans follow by their quantification using LC system. The optimal conditions of this UAE method were: aqueous EtOH 54.5% (v/v) as extraction solvent, with application of an ultrasound (US) frequency of 36.6 kHz during 60 min at 45 °C with a liquid to solid ratio of 25:1 mL/g dry weight (DW). Following its optimization using a full factorial design, the extraction method was validated according to international standards of the association of analytical communities (AOAC) to ensure precision and accuracy in the quantitation of each component of the SILM mixture. The efficiency of this UAE was compared with maceration protocol. Here, the optimized and validated conditions of the UAE allowed the highest extraction yields of SILM and its constituents in comparison to maceration. During UAE, the antioxidant capacity of the extracts was retained, as confirmed by the in vitro assays CUPRAC (cupric ion reducing antioxidant capacity) and inhibition of AGEs (advanced glycation end products). The skin anti-aging potential of the extract obtained by UAE was also confirmed by the strong in vitro cell-free inhibition capacity of both collagenase and elastase. To summarize, the UAE procedure presented here is a green and efficient method for the extraction and quantification of SILM and its constituents from the fruits of S. marianum, making it possible to generate extracts with attractive antioxidant and anti-aging activities for future cosmetic applications.

Silybum marianum (L.) Gaertn. is a well-known medicinal herb, primarily used in liver protection. Light strongly affects several physiological processes along with secondary metabolites biosynthesis in plants. Herein, S. marianum was exploited for in vitro potential under different light regimes in the presence of melatonin. The optimal callogenic response occurred in the combination of 1.0 mg/L α-naphthalene acetic acid and 0.5 mg/L 6-benzylaminopurine under photoperiod. Continuous light associated with melatonin treatment increased total flavonoid content (TFC), total phenolic content (TPC) and antioxidant potential, followed by photoperiod and dark treatments. The increased level of melatonin has a synergistic effect on biomass accumulation under continuous light and photoperiod, while an adverse effect was observed under dark conditions. More detailed phytochemical analysis showed maximum total silymarin content (11.92 mg/g dry weight (DW)) when placed under continuous light + 1.0 mg/L melatonin. Individually, the level of silybins (A and B), silydianin, isolsilychristin and silychristin was found highest under continuous light. Anti-inflammatory activities were also studied and highest percent inhibition was recorded against 15-lipoxygenase (15-LOX) for cultures cultivated under continuous light (42.33%). The current study helps us to better understand the influence of melatonin and different light regimes on silymarin production as well as antioxidant and anti-inflammatory activities in S. marianum callus extracts.

Painted nettle (Plectranthus scutellarioides (L.) R.Br.) is an ornamental plant belonging to Lamiaceae family, native of Asia. Its leaves constitute one of the richest sources of trans-rosmarinic acid, a well-known antioxidant and antimicrobial phenolic compound. These biological activities attract interest from the cosmetic industry and the demand for the development of green sustainable extraction processes. Here, we report on the optimization and validation of an ultrasound-assisted extraction (USAE) method using ethanol as solvent. Following preliminary single factor experiments, the identified limiting extraction parameters (i.e., ultrasound frequency, extraction duration, and ethanol concentration) were further optimized using a full factorial design approach. The method was then validated following the recommendations of the association of analytical communities (AOAC) to ensure the precision and accuracy of the method used to quantify trans-rosmarinic acid. Highest trans-rosmarinic acid content was obtained using pure ethanol as extraction solvent following a 45-minute extraction in an ultrasound bath operating at an ultrasound frequency of 30 kHz. The antioxidant (in vitro radical scavenging activity) and antimicrobial (directed toward Staphylococcus aureus ACTT6538) activities were significantly correlated with the trans-rosmarinic acid concentration of the extract evidencing that these key biological activities were retained following the extraction using this validated method. Under these conditions, 110.8 mg/g DW of trans-rosmarinic acid were extracted from lyophilized P. scutellarioides leaves as starting material evidencing the great potential of this renewable material for cosmetic applications. Comparison to other classical extraction methods evidenced a clear benefit of the present USAE method both in terms of yield and extraction duration.