Carduus edelbergii Rech. f. Mediated Fabrication of Gold Nanoparticles; Characterization and Evaluation of Antimicrobial, Antioxidant and Antidiabetic Potency of the Synthesized AuNPs.

Background: Due to the high expense, less effectiveness and more side effects of available synthetic medicine, the researchers and communities are focusing on phyto-based natural bioactive compounds, which are considered safer for the treatment of syndromes and chronic diseases. Aim: The current project was aimed to determine the phytochemicals constituents available in the aerial parts of methanol extract of Carduus edelbergii via GC-MS, fabrication of AuNPs mediated with the mentioned extract; characterization and evaluation of antimicrobial, antioxidant and antidiabetic potency of the synthesized AuNPs. Methods: Confirmation of green synthesis of AuNPs, functional groups responsible for the reduction in Au+, size and crystallinity, morphology and quantity of gold (Au) were carried out by Ultraviolet-Visible (UV-Vis) spectroscopy, Transform Infrared (FTIR) spectroscopy, Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD) and dispersive X-ray (EDX), respectively, whereas in vitro antioxidant characteristics were assessed by DPPH and ABTS assays. Wistar albino rats were used to test the anti-diabetic properties of the methanol extract and AuNPs. Results: GC-MS revealed that the diluted methanol extract of Carduus edelbergii consists of about 19 chemical constituents. Among the identified compounds, the 13-Docosenoic acid, methyl ester, (Z)­has the highest concentration (38.16%), followed by 9-Octadecenoic acid, methyl ester, (E)­(15.72%) and n-Hexadecanoic acid (15.07%). Methanol extract and its fabricated nanoparticles showed significant antioxidant and antimicrobial activities. In vivo antidiabetic study revealed a noteworthy (p < 0.05) decline in body weight and HDL and elevated concentration of blood glucose, bilirubin, creatinine, urea, triglyceride, VLDL, LDL, ALP, ALT and AST in diabetic control. The said changes were recovered significantly (p < 0.05) by treatment of diabetic rats with methanol extract (150 and 300 mg/Kg BW) and AuNPs of Carduus edelbergii (5 and 10 mg/Kg BW). Conclusion: The green synthesized AuNPs exhibit significant antioxidant, antimicrobial and antidiabetic characteristics.

Phenolic Composition of Inflorescences of Carduus nutans L.

Carduus nutans L. (Compositae) has been extensively used for medicinal purposes. As other representatives of the genus it is implemented in the treatment of liver disorders and used as diuretic and digestive agent. Previous studies have determined the predominant classes of secondary metabolites in Carduus species. Among the major representatives of their extracts flavonoids, phenolic acids, coumarins, sterols and terpenes were identified. The antiradical capacity of three extracts assessed in the DPPH test revealed the highest radical scavenging properties of methanol extract (the EC50 618±10.03 µg/mL) and based on these results it was selected for phenolic content determination. (TPC=61.49 mg/g). To understand better the induced pharmacological effects of the plant the aim of the study was to determine the composition of the methanol extract from the inflorescences of C. nutans and to study their phenolic composition. In order to determine the composition in a more efficient way, the crude extract was fractionated and subjected to hydrolyses. As a result more than twenty phenolic acids and flavonoids were identified in the extract and fractions by HPLC-DAD and/or HPLC-ESI-TOF-MS. The total extract was later subjected to fractionation by centrifugal partition chromatography using the Arizona system composed of hexane/ethyl acetate/methanol/water (0.7 : 4 : 0.8 : 4 v/v/v/v) to produce fractions enriched in flavonoids that are of high pharmacological significance.

Determination of biological activity of Carduus lanuginosus: an endemic plant in Turkey.

The genus Carduus is traditionally used in the Anatolian folk medicine for treating various diseases. Therefore, the enzyme inhibiting potential, antioxidant-antimicrobial activity, and phytochemical profile of Carduus lanuginosus extracts were investigated. The analysis of phenolic compounds was carried out by using RP-HPLC for the chemical characterization of methanol extract. The total polyphenols, total phenolic and flavonoid contents, antioxidant activity (ABTS and DPPH assay), α-amylase, and α-glucosidase inhibition activities were determined using colorimetric methods. Moreover, the antimicrobial activity was examined using the disc diffusion and microdilution methods. The ethylacetate extract was found to have the highest flavonoid and phenolic content. The water and hexane extracts showed strong enzyme inhibitory activity against the α-amylase and α-glucosidase. The methanol extract was found to contain high concentration of chlorogenic acid. The hexane and ethylacetate extracts showed to have significant MIC values on Enterococcus faecium. In conclusion, the extracts of C. lanuginosus might have a significant potential for the use as a natural pharmaceutical agent.

Bioactive potential of Indian stinging plants leaf extract against pathogenic fungi.

Background & methods We investigated the in vitro antioxidant and antifungal activity by agar disc diffusion assay of leaf extract of some stinging plants namely, Urtica dioica L., Tragia involucrate L., Carduus nutans L. and Mucuna pruriens (L.) DC., against pathogenic fungi causing infections/diseases. Results M. pruriens (Disc 4), T. involucrate (Disc 2), U. dioica (Disc 1) showed significant antifungal activity against all tested pathogens, while C. nutans (Disc 3) showed intermediate activity against only Chaetomium globosum (Cg). The leaf extract of M. Pruriens showed maximum total phenol content (~1004 µg g-1 dry wt) followed by T. involucrate, C. nutans and U. dioica. However, the ascorbate was observed highest in T. involucrate (~10.3 µg g-1 dry wt) followed by M. pruriens (~9.2 µg g-1 dry wt) but the difference was not significant (p ≤ 0.05). Likewise, M. pruriens showed maximum anthocyanin content (~0.3 µg g-1 dry wt). The activity of antioxidant enzymes revealed that M. Pruriens showed maximum ascorbate peroxidase (APX) activity, while the highest guaiacol peroxidase (GPX) and catalase (CAT) activities were observed in C. nutans and U. dioica, respectively. Conclusions M. Pruriens showed potential in vitro antioxidant and antifungal activity against studied pathogens that may be used for ethno-pharmacological uses.

Characterization of four wild edible Carduus species from the Mediterranean region via phytochemical and biomolecular analyses.

Carduus species (Compositae) are widely distributed in the Mediterranean area, and traditionally used for both food and medicinal purposes. The hydroalcoholic extracts of four wild edible Carduus species collected in Sardinia (Carduus argyroa Biv., Carduus nutans subsp. macrocephalus (Desf.) Nyman, Carduus pycnocephalus L., Carduus cephalanthus Viv.) were analyzed and characterized by HPLC-PDA-MS/MS and PCR-RFLP of the nrDNA internal transcribed spacer (ITS). Flavonoids and caffeoylquinic acid derivatives were the predominant classes of secondary metabolites characterizing the extracts. The ITS region was sequenced in parallel, and a PCR-RFLP method was applied with three selective restriction enzymes. Statistical analyses, on both chemical and biomolecular results, revealed that individuals clustered according to their taxonomic classification. The combination of the two techniques discriminates the four species within the genus, giving further information on these little-investigated plants, traditionally used in the Mediterranean area and in Sardinia.

Optimized conversion of antiproliferative lignans pinoresinol and epipinoresinol: Their simultaneous isolation and identification by centrifugal partition chromatography and high performance liquid chromatography.

High amount of the valuable lignan pinoresinol (PR) was determined in Carduus nutans fruit (7.8mg/g) for the first time. A preparative separation method using two consecutive, identical steps of centrifugal partition chromatography (CPC) was developed in order (i) to isolate PR and (ii) to subsequently isolate PR and its 7' epimer epipinoresinol (EPR) simultaneously after an optimized acid treatment which resulted in PR epimerization forming equal amounts of PR and EPR, from C. nutans fruit. As optimal conditions, a two-phase solvent system consisting of methyl tert-butyl ether:acetone:water (4:3:3, v/v/v) for CPC separation, and an acid treatment performed at 50°C for 30min for the epimerization were applied. Thus, 33.7mg and 32.8mg PR and EPR, in as high as 93.7% and 92.3% purity, were isolated from 10.0gC. nutans fruit, representing 86.4% and 84.1% efficiency, respectively. Conversion characteristic of PR and EPR in acidic medium, determined as a function of time and temperature of acid treatment provides their unambiguous identification by on-line high performance liquid chromatography (HPLC). Antiproliferative assay of isolated PR and EPR in two different types of colon cancer cell lines (HCT116 and SW480) confirmed that both epimers caused a more significant decrease of viability in HCT116 cells than in SW480 cells, suggesting their similar mechanism of antiproliferative action.

Stinging plants: as future bio-weapon.

In the present opinion paper, we have been introducing for the first time the stinging plants and/or their biological toxins as novel bio-threat agents that may be used for the development of bio-weapons for self-defence purpose. The selected studied stinging plants are having dual role as nutraceutical and ethno-pharmacological uses apart from their less explored stinging property. However, future detailed work is required for identification and characterization of the precise stinging chemical components that will be used for the formulation of novel bio-warfare agents for self-defence purpose.

Two new glycosides from Carduus acanthoides.

Two new glycosides, syringic acid-4-O-ß-L-arabinopyranoside and kaempferol-3-O-α-L-rhamnopyranosyl-7-O-ß-D-glucuronopyranoside, were isolated from whole plants of Carduus acanthoides (Asteraceae), and their structures were elucidated on the basis of spectroscopic analysis.

[Chemical constituents from whole plants of Carduus acanthoides].

Fourteen compounds were isolated from wholeplants of Carduus acanthoides by various chromatographic techniques including column chromatography over HP-20 macroporous resin, MCI gel, silica gel, Sephadex LH-20 and ODS and reversed-phase HPLC. Their structures were identified as salidroside (1), 2-(3,4-dihydroxyphenyl)-ethyl-O-beta-D-glucopyranoside (2), 3,5-di-hydroxyphenethyl alcohol-3-O-beta-D-glucopyranoside (3), p-coumaric acid (4), 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl) propan-1-one (5), 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl) propan-1-one (6), syringin (7), p-hydroxybenzaldehyde (8), salicylic acid (9), tachioside (10), vanillic acid-4-O-beta-D-glucopyranoside (11), syringic aldehyde (12), 2,6-dimethoxy-4-hydroxyphenol-1-O-beta-D-glucopyranoside (13), and 2, 6-dimethoxy-p-hydroquinone-4-0-P-D-glucopyranoside (14) on the basis of spectroscopic data analysiS. All compounds were isolated from the genus Carduus for the first time except for compounds 4 and 7.

Inhibition of key enzymes linked to obesity by preparations from Mediterranean dietary plants: effects on α-amylase and pancreatic lipase activities.

One of the most important strategy in the treatment of obesity includes the development of nutrient digestion and absorption inhibitors. Inhibition of digestive enzymes is one of the most widely studied mechanisms used to determine the potential efficacy of natural products as hypolipidemic and hypoglycaemic agents. In vitro studies here reported were performed to evaluate the inhibitory activity of five species(as hydroalcoholic extracts) of edible plants from Calabria region (Italy) on amylase and lipase by monitoring the hydrolysis of p-NPC and the hydrolysis of glycoside bonds indigestible carbohydrate foods. The formulation obtained from Clematis vitalba L. exhibited the strongest inhibitory effect on pancreatic lipase (IC50=0.99 mg/ml) and on α-amylase(IC50=31.52 µg/ml). In order to explore metabolome production HPTLC analysis of the extracts was performed, revealing the predominance of (±)-catechin, caffeic acid and chlorogenic acid in C. vital ba formulation at concentration of 23.18±3.14,13.63±0.65 and 18.88±0.76 mg/g, respectively. GC/MS analysis was used to identify fatty acids and terpene composition.

Chemical constituents from whole plants of Carduus acanthoides / 中国中药杂志

Fourteen compounds were isolated from wholeplants of Carduus acanthoides by various chromatographic techniques including column chromatography over HP-20 macroporous resin, MCI gel, silica gel, Sephadex LH-20 and ODS and reversed-phase HPLC. Their structures were identified as salidroside (1), 2-(3,4-dihydroxyphenyl)-ethyl-O-beta-D-glucopyranoside (2), 3,5-di-hydroxyphenethyl alcohol-3-O-beta-D-glucopyranoside (3), p-coumaric acid (4), 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl) propan-1-one (5), 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl) propan-1-one (6), syringin (7), p-hydroxybenzaldehyde (8), salicylic acid (9), tachioside (10), vanillic acid-4-O-beta-D-glucopyranoside (11), syringic aldehyde (12), 2,6-dimethoxy-4-hydroxyphenol-1-O-beta-D-glucopyranoside (13), and 2, 6-dimethoxy-p-hydroquinone-4-0-P-D-glucopyranoside (14) on the basis of spectroscopic data analysiS. All compounds were isolated from the genus Carduus for the first time except for compounds 4 and 7.

Protection of apigenin against kainate-induced excitotoxicity by anti-oxidative effects.

Apigenin (5,7,4'-trihydroxyflavone) is a principal ingredient of Cirsium japonicum. These experiments were performed to determine whether apigenin has neuroprotective effects against kainic acid (KA)-induced excitotoxicity in vitro and in vivo. Intraperitoneal (i.p.) administration of apigenin (25, 50 mg/kg) decreased the seizure scores induced by KA injection (40 mg/kg, i.p.) in mice. In addition, the convulsion onset time was significantly delayed by apigenin administration. Moreover, we found that apigenin blocked KA-induced seizure-form electroencephalogram (EEG) discharge activity in the brain cortex. In hippocampal cells, apigenin inhibited KA-induced excitotoxicity in a dose-dependent manner as measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To study the possible mechanisms underlying the in vitro neuroprotective effects of apigenin against KA-induced cytotoxicity, we also examined the effect of apigenin on intracellular reactive oxygen species (ROS) elevations in cultured hippocampal neurons and found that apigenin treatment dose-dependently inhibited intracellular ROS elevation. The remarkable reduction of glutathione (GSH) levels induced by KA in hippocampal tissues was reversed by apigenin in a dose-dependent manner. In addition, similar results were obtained after pretreatment with free radical scavengers such as trolox and dimethylthiourea (DMTU). Finally, after confirming the protective effect of apigenin in hippocampal CA3 region, we found apigenin is an active compound in KA-induced neuroprotection. These results collectively indicate that apigenin alleviates KA-induced excitotoxicity by quenching ROS as well as inhibiting GSH depletion in hippocampal neurons.

Simultaneous determination of bioactive flavonoids in some selected Korean thistles by high-performance liquid chromatography.

In order to facilitate the quality control of some selected Korean thistles (Cirsii Herb), Cirsium japonicum var ussuriense, C. japonium var spinosissimum, C. setidens, C. pendulum, C. nipponicum, Carduus crispus, and Breea segetum, a simple, accurate and reliable high performance liguid chromatography method was developed for the simultaneous determination of the six flavonoids: luteolin 5-O-glucoside (1), luteolin 7-O-glucoside (2), hispidulin 7-O-neohesperidoside (3), luteolin (4), pectolinarin (5), and apigenin (6), which were selected as the chemical markers of the thistles. Separation was achieved on an Agilent Eclipse XDB-C18 column with a gradient solvent system of 0.1% trifluoroacetic acid aqueous-methanol at a flow-rate of 1.0 mL/min and detected at 254 nm. All six calibration curves showed good linearity (R(2) > 0.991). The method was reproducible with intra- and inter-day variations of less than 6%. The recoveries were in the range of 90.01-100.05%. This analysis method was successfully utilized to quantify the six flavonoids in the 22 batches of the thistles. The results demonstrated that this method is simple, reliable and suitable for the quality control of this medicinal herb.

Counting pollen grains using readily available, free image processing and analysis software.

BACKGROUND AND AIMS: Although many methods exist for quantifying the number of pollen grains in a sample, there are few standard methods that are user-friendly, inexpensive and reliable. The present contribution describes a new method of counting pollen using readily available, free image processing and analysis software. METHODS: Pollen was collected from anthers of two species, Carduus acanthoides and C. nutans (Asteraceae), then illuminated on slides and digitally photographed through a stereomicroscope. Using ImageJ (NIH), these digital images were processed to remove noise and sharpen individual pollen grains, then analysed to obtain a reliable total count of the number of grains present in the image. A macro was developed to analyse multiple images together. To assess the accuracy and consistency of pollen counting by ImageJ analysis, counts were compared with those made by the human eye. KEY RESULTS AND CONCLUSIONS: Image analysis produced pollen counts in 60 s or less per image, considerably faster than counting with the human eye (5-68 min). In addition, counts produced with the ImageJ procedure were similar to those obtained by eye. Because count parameters are adjustable, this image analysis protocol may be used for many other plant species. Thus, the method provides a quick, inexpensive and reliable solution to counting pollen from digital images, not only reducing the chance of error but also substantially lowering labour requirements.

Changes in microbial community structure, methanogenesis and rumen fermentation in response to saponin-rich fractions from different plant materials.

AIMS: Investigation of the effects of saponin-rich fractions on rumen fermentation, methane production and the microbial community. METHODS AND RESULTS: Saponins were extracted from Carduus, Sesbania and Knautia leaves and fenugreek seeds. Two levels of saponin-rich fractions with a substrate were incubated using the Hohenheim gas method. Methane was measured using an infrared-based methane analyser and microbial communities using quantitative PCR. On addition of saponin-rich fractions, methane and short-chain fatty acid production was not affected. The protozoal counts decreased by 10-39%. Sesbania saponins decreased methanogen population by 78%. Decrease in ruminal fungal population (20-60%) and increase in Fibrobacter succinogenes (21-45%) and Ruminococcus flavefaciens (23-40%) were observed. CONCLUSIONS: The saponins evaluated possessed anti-protozoal activity; however, this activity did not lead to methane reduction. Fenugreek saponins seemed to have potential for increasing rumen efficiency. The saponins altered the microbial community towards proliferation of fibre-degrading bacteria and inhibition of fungal population. SIGNIFICANCE AND IMPACT OF THE STUDY: The uni-directional relationship between protozoal numbers and methanogenesis, as affected by saponins, is not obligatory. All saponins might not hold promise for decreasing methane production from ruminants.

Comparative antioxidant activity and HPLC profiles of some selected Korean thistles.

As yet, no comparative analyses have been conducted regarding the comparative antioxidant activities and HPLC profiles of thistles distributed in Korea. Thus, this study was performed in order to evaluate the antioxidant potentials of seven Korean thistles: Cirsium lineare, Cirsium chanroenicum, Cirsium setidens, Cirsium japonicum var. ussuriense, Cirsium nipponicum, Cirslum pendulum and Carduus crispus, via peroxynitrite and DPPH free radical assays. Among seven Korean thistles, Carduus crispus exhibited the most significant antioxidant activity in both DPPH assay and peroxynitrite. In order to characterize the compounds contained in Korean thistles, we conducted HPLC analyses on the following ten flavonoids: luteolin-5-glucoside (1), luteolin-7-glucoside (2), apigenin-7-glucoside (3), hispidulin-7-neohesperidoside (4), apigenin-7-glucuronide (5), cirsimarin (6), pectolinarin (7), luteolin (8), apigenin (9) and acacetin (10). The results of our HPLC analyses indicated the presence of pectolinarin in the whole plants of C. setidens, C. lineare, C. nipponicum, C. pendulum, the aerial and underground parts of C. japonicum var. ussuriense, and the aerial parts of C. chanroenicum. Moreover, we were able to identify hispidulin-7-neohesperidoside and luteolin-7-glucoside in the whole plants of Carduus crispus, acacetin in the aerial parts of C. chanroenicum, cirsimarin in C. lineare.

[Studies on the chemical constituents of Carduus crispus L].

OBJECTIVE: To study the chemical constituents of Carduus crispus. METHOD: Using chromatographic methods to isolate compounds and using chemical and spectral methods to elucidate the structures of the isolated compounds. RESULT: Eight compounds were elucidated as beta-amyrin palmitate, taraxastery acetate, luteolin-7-O-alpha-L-rhamanopyranosyl-(1-->2)-beta-D-glucopyranoside, luteolin-7-O-beta-D-glucopyranoside, triacontanic acid, beta-sitosterol, stigmasterol and stigmast-7-en-3 beta-ol. CONCLUSION: All the compounds were obtained from this plant for the first time.