Wild Vitex agnus-castus L.: Phytochemical Characterization, Acute Toxicity, and Bioactive Properties.

Wild Vitex agnus-castus (VAC) is a Mediterranean plant that is rich in bioactive metabolites. This study aimed to validate, for the first time, the beneficial use of VAC fruits and fruit decoctions (VFDs) through in vitro and in vivo trials. Forty-one volatile components were detected in VAC fruits, with 1,8-cineole (30.3%) comprising the majority. The antioxidant activity of VFD was measured by using different in vitro methods (EC50 of 0.16 mg/mL by ß-carotene bleaching inhibition assay) and by measuring the DNA protection power. Using the disc diffusion assay, the antimicrobial activity of VFD was evaluated, and it exhibited a noticeable anticandidal activity. VFD did not cause any toxicity or mortality in rats treated with doses > 200 mg/kg. Using the acetic acid writhing test, the antinociceptive activity of VFD was measured. Our results showed that VFD at 200 mg/kg exhibited a higher analgesic activity (81.68%) than acetylsalicylic acid used as a positive control (74.35%). Its gastroprotective ability was assessed by HCl/ethanol-induced gastric lesions, which were remarkably inhibited (84.62%) by intraperitoneal administration of VFD. This work helps to validate the popular use of VAC to treat nociceptive, inflammatory, and gastric disorders and encourages researchers to further investigate the identification of pharmacological compounds from this species.

Bioactivities and in silico study of Pergularia tomentosa L. phytochemicals as potent antimicrobial agents targeting type IIA topoisomerase, TyrRS, and Sap1 virulence proteins.

Pergularia tomentosa L. (P. tomentosa) has been largely used in Tunisian folk medicine as remedies against skin diseases, asthma, and bronchitis. The main objectives of this study were to identify phytochemical compounds that have antioxidant and antimicrobial properties from the stem, leaves, and fruit crude methanolic extracts of P. tomentosa, and to search for tyrosyl-tRNA synthetase (TyrRS), topoisomerase type IIA, and Candidapepsin-1 (SAP1) enzyme inhibitors through molecular docking study. Phytochemical quantification revealed that fruit and leaves extracts displayed the highest total flavonoids (582 mg QE/g Ex; 219 mg QE/g Ex) and tannins content (375 mg TAE/g Ex; 216 mg TAE/g Ex), also exhibiting significant scavenging activity to decrease free radicals for ABTS, DPPH, ß-carotene, and FRAP assay with IC50 values (> 1 mg/mL). Additionally, promising antimicrobial activities towards different organs have been observed against several bacteria and Candida strains. From the liquid chromatography-mass spectrometry (LC-MS) analysis, five polyphenolic compounds, namely digitoxigenin, digitonin glycoside and calactina in the leaves, kaempferol in the fruit, and calotropagenin in the stems, were identified. They were also analyzed for their drug likeliness, based on computational methods. Molecular docking study affirmed that the binding affinity of calactin and actodigin to the active site of TyrRS, topoisomerase type IIA, and SAP1 target virulence proteins was the highest among the examined dominant compounds. Therefore, this study indicated that P. tomentosa methanolic extracts displayed great potential to become a potent antimicrobial agent and might be a promising source for therapeutic and nutritional functions. These phytocompounds could be further promoted as a candidate for drug discovery and development.

Nutritional properties, identification of phenolic compounds, and enzyme inhibitory activities of Feijoa sellowiana leaves.

The nutritional values, the identification of individual phenolic compounds as well as the functional properties (in vitro acetylcholinesterase (AChE) and pancreatic lipase inhibition) of Tunisian Feijoa sellowiana (O.Berg) leaves were determined by high-performance liquid chromatography coupled with diode array and mass spectrometry detector. Feijoa Sellowiana  leaves Extract (FSLE) were evaluated in the present work. Macronutrient components (proteins, carbohydrates, dietary fiber, lipids, and ashes) of FSLE as well as their fatty acids and mineral content were determined. The individual phenolic composition was characterized by the presence of flavonols (mostly quercetin glycosides), flavan-3-ols (procyanidins and catechins), and phenolic acid derivatives. To understand the possible inhibition mechanism exerted by the extract components, enzyme kinetic studies using derived Lineweaver-Burk (LB) plots and Michaelis-Menten were performed. The obtained results evidenced that FSLE exhibited important AChE inhibitory effect, behaving as a noncompetitive inhibitor. Furthermore, the leaves' extract inhibited irreversibly the pancreatic lipase, with maximum inhibition (70%) achieved at 1 mg/ml. PRACTICAL APPLICATIONS: In many countries of the world, Feijoa Sellowiana leaves are used as an alternative to tea drinks and as a folkloric medicine. Moreover, it is well known that the phytochemical compounds are a practical alternative for the prevention of many chronic diseases such as Alzheimer's and obesity. For this reason, the present study aimed to evaluate the nutritional, individual phenolic compounds, and functional properties of Feijoa Sellowiana leaves to valorize them as a food and pharmaceutical bioactive human source.

Phytoestrogens inhibit key-enzymes linked to obesity, type 2 diabetes and liver-kidney toxicity in high fructose-fat diet in mice.

Objective: To evaluate the effect of the administration of phytoestrogens on obesity, type 2 diabetes, and liver-kidney toxicity. Methods: Phytoestrogens (phyto(E2)) were administrated to high fructose-fat diet (HFFD). Results: This study showed that administration of phyto(E2) to HFFD-mice inhibited lipase activity by 34%, decreased body weight by 20% and modulated lipid profile, showed a decrease in total-cholesterol (TC) and LDL-cholesterol (LDL-C) rates in the plasma by 59% and 42%, respectively, and increased the HDL-cholesterol (HDL-C) level by 31%. In addition, the administration of phytoestrogens to HFFD-mice exerts an inhibitory effect on α-amylase activity and decreased glucose level by 28% and increase in liver glycogen level by 33%; and ameliorate oral glucose tolerance test. Conclusions: This study demonstrate that phyto(E2) has both a promising potential with regards to the inhibition of intestinal lipase and α-amylase activities, and a valuable hypoglycemic and hypolipidemic function.

Rhaponticum acaule (L) DC essential oil: chemical composition, in vitro antioxidant and enzyme inhibition properties.

BACKGROUND: α-glucosidase is a therapeutic target for diabetes mellitus (DM) and α-glucosidase inhibitors play a vital role in the treatments for the disease. Furthermore, xanthine oxidase (XO) is a key enzyme that catalyzes hypoxanthine and xanthine to uric acid which at high levels can lead to hyperuricemia which is an important cause of gout. Pancreatic lipase (PL) secreted into the duodenum plays a key role in the digestion and absorption of fats. For its importance in lipid digestion, PL represents an attractive target for obesity prevention. METHODS: The flowers essential oil of Rhaponticum acaule (L) DC (R. acaule) was characterized using gas chromatography-mass spectrometry (GC-MS). The antioxidant activities of R. acaule essential oil (RaEO) were also determined using 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), reducing power, phosphomolybdenum, and DNA nicking assays. The inhibitory power of RaEO against α-glucosidase, xanthine oxidase and pancreatic lipase was evaluated. Enzyme kinetic studies using Michaelis-Menten and the derived Lineweaver-Burk (LB) plots were performed to understand the possible mechanism of inhibition exercised by the components of this essential oil. RESULTS: The result revealed the presence of 26 compounds (97.4%). The main constituents include germacrene D (49.2%), methyl eugenol (8.3%), (E)-ß-ionone (6.2%), ß-caryophyllene (5.7%), (E,E)-α-farnesene (4.2%), bicyclogermacrene (4.1%) and (Z)-α-bisabolene (3.7%). The kinetic inhibition study showed that the essential oil demonstrated a strong α-glucosidase inhibiton and it was a mixed inhibitor. On the other hand, our results evidenced that this oil exhibited important xanthine oxidase inhibitory effect, behaving as a non-competitive inhibitor. The essential oil inhibited the turkey pancreatic lipase, with maximum inhibition of 80% achieved at 2 mg/mL. Furthermore, the inhibition of turkey pancreatic lipase by RaEO was an irreversible one. CONCLUSION: The results revealed that the RaEO is a new promising potential source of antioxidant compounds, endowed with good practical applications for human health.

Antimicrobial Activity of Scabiosa arenaria Forssk. Extracts and Pure Compounds Using Bioguided Fractionation.

The emergence of multidrug resistant pathogens threatened the clinical efficacy of many existing antibiotics. This situation has been recognized globally as a serious concern and justifies further research to discover antimicrobial agents from natural origins including plant extracts. The aim of our work was to evaluate the antimicrobial activities of Scabiosa arenaria Forssk. extracts and pure compounds using a bioguided fractionation, and try to explain some traditional use of this genus. The best antimicrobial activity-guided fractionation was obtained by BuOH fractions of flowers, fruits and (stems and leaves) against Escherichia coli, Pseudomonas aeruginosa and Candida albicans with minimum inhibitory concentration (MIC) values from 0.0195 to 5 mg/ml. Escherichia coli was the most affected bug, thus the MIC of fruits BuOH extract showed the best anti-Escherichia coli activity (MIC = 0.0195 mg/ml), followed by the (stems and leaves) and flowers BuOH extracts; MIC = 0.078 and 0.15 mg/ml, respectively. Furthermore, the subfractions obtained from these three mixed fractions showed also an important antimicrobial activity against the three microorganisms, with MIC values between 0.0195 and 0.312 mg/ml. The fractionation of the aerial part BuOH fraction led to the isolation of oleanolic acid (1) and luteolin 7-O-glucopyranoside (2) which are reported here for the first time from S. arenaria. Both compounds showed good antimicrobial activities with MIC values ranging from 170 to 683 µm and 86 to 347 µm, respectively. These results support the use of the Scabiosa genus to inhibit the growth of tested pathogenic bacteria and yeasts which may reduce illnesses associated with their exposure.

Characterisation of phenolic antioxidants in Scabiosa arenaria flowers by LC-ESI-MS/MS and NMR.

OBJECTIVES: This work describes the bioguided fractionation of the flower's ethyl acetate fraction of Scabiosa arenaria Forssk. (Dipsacaceae). METHODS: The identification of the pure compound isolated has been studied by mono-dimensional NMR experiments. The mixture of phenolic compounds was analysed by LC-ESI-MS/MS. The antioxidant activity has been evaluated by the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. KEY FINDINGS: The bioguided fractionation of the flower's ethyl acetate fraction of Scabiosa arenaria led to the isolation of a pure compound: luteolin. The mixture of three phenolic compounds was identified as: 1, 4-O-dicaffeoylquinic acid, apigenin 7-O-glucoside and luteolin-7-O-glucoside. Two of which are reported here for the first time in Scabiosa genus. Luteolin had the highest antioxidant activity with an IC50 value of 0.02 ± 0.007 mg/ml, followed by the three phenolic compounds with an IC50 value of 0.025 ± 0.008 mg/ml. CONCLUSIONS: The results of the present work indicate that S. arenaria flower's ethyl acetate extract could be used as natural antioxidant agents in food preservation and human health.

Chemical composition and biological activities of Eruca vesicaria subsp. longirostris essential oils.

Context To date, there are no reports to validate the Tunisian traditional and folklore claims of Eruca vesicaria (L) Cav. subsp. longirostris (Brassicaceae) for the treatment of disease. Objective Investigation of the chemical composition antimicrobial and antioxidant activity of essential oils from Eruca longirostris leaves, stems, roots and fruits. Materials and methods The essential oils of E. longirostris from leaves, stems, roots and fruits were obtained after 4 h of hydrodistillation. Chemical compositions were determined using a combination of GC/FID and GC/MS. The in vitro antimicrobial activity of the volatile constituents of E. longirostris was performed in sterile 96-well microplates against three Gram-positive, four Gram-negative bacteria and one strain as yeast. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration values were reported. Furthermore, the antioxidant activity was evaluated by DPPH and ABTS assays. Results The main compound for fruits, stems and roots was the erucin (96.6%, 85.3% and 83.7%, respectively), while ß-elemene (35.7%), hexahydrofarnesylacetone (23.9%), (E)-ß-damascone (15.4%), erucin (10.6%) and α-longipinene (9.6%) constituted the major compounds in the essential oil of the leaves. The experimental results showed that in all tests, essential oil of fruits showed the better antioxidant activity than the others. On the other hand, the oils of stems, fruits and roots showed significant antimicrobial activity with MIC values ranging from 0.125 to 0.31 mg/mL against Candida species, Gram-positive and Gram-negative bacteria, mainly Salmonella enterica. Conclusions The present results indicate that essential oils of E. longirostris can be used as a source of erucin.

α-Glucosidase inhibition by Tunisian Scabiosa arenaria Forssk. extracts.

Recent decades have witnessed a sharp increase in the incidence and prevalence of type 2 diabetes mellitus. One antidiabetic therapeutic approach is to reduce gastrointestinal glucose production and absorption through the inhibition of carbohydrate-digesting enzymes such as α-amylase and α-glucosidase. In this study, crude extracts and their corresponding fractions of flowers, fruits, (stems and leaves) and roots of the endemic North African plant Scabiosa arenaria Forssk. were screened for their ability of α-glucosidase inhibition. It was found that the fruits ethyl acetate (EtOAc), the fruits butanolic (n-BuOH) and the flowers ethyl acetate (EtOAc) fractions inhibited α-glucosidase in a non competitive manner with IC50 values of 0.11±0.09, 0.28±0.04 and 0.221±0.01mg/ml, respectively. RP-HPLC analysis indicated that the major components of these active fractions are flavonoid aglycone, cinnamic acid and its derivatives. This result supports the conclusion that the three studied fractions could be a useful natural source for the development of a novel α-glucosidase inhibitory agent against diabetic complications.

Phenolic composition, antioxidant and anti-acetylcholinesterase activities of the Tunisian Scabiosa arenaria.

CONTEXT: There is a need for the discovery of novel natural antioxidants and acetylcholinesterase inhibitors (AChEIs) that are safe and effective at a global level. This is the first study on antioxidant and anti-acethylcholinesterase activity of Scabiosa arenaria Forssk (Dipsacaceae). OBJECTIVE: The antioxidant potential and anti-acetylcholinesterase (AChE) activity of S. arenaria were investigated. MATERIAL AND METHODS: The crude, ethyl acetate (EtOAc), butanol (n-BuOH) and water extracts prepared from flowers, fruits and stems and leaves of S. arenaria were tested to determine their total polyphenol content (TPC), total flavonoid content (TFC), total condensed tannin content (CTC) and their antioxidant activity by using 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), reducing power and ß-carotene bleaching inhibition activity. Anti-AChE activity was also determined. RESULTS: EtOAc and n-BuOH fractions of fruits had both the highest (TPC) (269.09 mg gallic acid equivalents/g dry weight). The crude extract of stems and leaves had the highest TFC (10.9 mg quercetin equivalent/g dry weight). The n-BuOH fraction of stems and leaves had the highest CTC (489.75 mg catechin equivalents/g dry weight). The EtOAc fraction of flowers exhibit a higher activity in each antioxidant system with a special attention for DPPH assay (IC50 = 0.017 mg/mL) and reducing power (EC50 = 0.02 mg/mL). The EtOAc and n-BuOH fractions of stems and leaves showed strong inhibition of AChE (IC50 = 0.016 and 0.029 mg/mL, respectively). DISCUSSION AND CONCLUSIONS: These results suggest the potential of S. arenaria as a possible source of novel compounds and as an alternative antioxidant and AChEIs.

Partial characterization of a novel amylase activity isolated from Tunisian Ficus carica latex.

CONTEXT: A large number of plants still need to be investigated through screening of amylases suitable for industry. In the present study, and for the first time, we describe the amylolytic activity of Saint Pedro Ficus carica L. (Moraceae) crude latex of Kahli and Bidhi varieties. OBJECTIVE: Effects of temperature, pH, metal ions, and inhibitors and compatibility with some commercial detergents were investigated for amylase activity. MATERIALS AND METHODS: Amylase activity was screened in crude latex using the DNS method and potato starch as a substrate. Analyses of amylolytic reaction products by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were performed. RESULTS: Bidhi and Kahli amylases were active in optimal pH of 6.5 and 7 at 45°C, respectively, displaying a half life of 85 and 60 min, respectively, at 80°C, and they were very stable in a wide range of pH (4-12). Bidhi amylase activity increased to 260% by addition of 10(-3) mM Fe(2+) or 10(-2) mM Cu(2+), and was strongly inhibited by Mg(2+) and EDTA. In the presence of Ca(2+) and Mg(2+), Kahli amylase activity was dramatically enhanced by 220 and 260%, respectively. The compatibility of both amylases with certain commercial detergents was also shown to be good as enzymes retained up to 98% of their activities after 30 min of incubation at 80°C. DISCUSSION AND CONCLUSION: Analysis of amylolytic reaction products by TLC and HPLC suggested that Kahli amylase was an amyloglucosidase and Bidhi amylase was ß-fructose, α(1-4) glucose. Bidhi amylase is a good choice for application in starch, food, detergents and medical industries.