A Series of New Hydrazone Derivatives: Synthesis, Molecular Docking and Anticholinesterase Activity Studies.

BACKGROUND: Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are known to be serine hydrolase enzymes responsible for the hydrolysis of acetylcholine (ACh), which is a significant neurotransmitter for regulation of cognition in animals. Inhibition of cholinesterases is an effective method to curb Alzheimer's disease, a progressive and fatal neurological disorder. OBJECTIVE: In this study, 30 new hydrazone derivatives were synthesized. Then we evaluated their anticholinesterase activity of compounds. We also tried to get insights into binding interactions of the synthesized compounds in the active site of both enzymes by using molecular docking approach. METHODS: The compounds were synthesized by the reaction of various substituted/nonsubstituted benzaldehydes with 6-(substitute/nonsubstituephenyl)-3(2H)-pyridazinone-2-yl propiyohydrazide. Anticholinesterase activity of the compounds was determined using Ellman's method. Molecular docking studies were done by using the ADT package version 1.5.6rc3 and showed by Maestro. RMSD values were obtained using Lamarckian Genetic Algorithm and scoring function of AutoDock 4.2 release 4.2.5.1 software. RESULTS: The activities of the compounds were compared with galantamine as cholinesterase enzyme inhibitor, where some of the compounds showed higher BChE inhibitory activity than galantamine. Compound F111 was shown to be the best BChE inhibitor effective in 50 µM dose, providing 89.43% inhibition of BChE (IC50=4.27±0.36 µM). CONCLUSION: This study supports that novel hydrazone derivates may be used for the development of new BChE inhibitory agents.

Drug Design of Inhibitors of Alzheimer's Disease (AD): POM and DFT Analyses of Cholinesterase Inhibitory Activity of ß-amino di-Carbonyl Derivatives.

BACKGROUND: Since deficit of acetylcholine has been evidenced in the Alzheimer's disease (AD) patients, cholinesterase inhibitors are currently the most specified drug category for the remediation of AD. METHOD: In the present study, 16 compounds (1-16) with dicarbonyl skeletons have been synthesized and tested for their inhibitory potential in vitro against AChE and BChE using ELISA microtiter plate assays at 100 µg/mL. Since metal accumulation is related to AD, the compounds were also tested for their metal-chelation capacity. RESULTS AND CONCLUSION: All the investigated dicarbonyl compounds exerted none or lower than 30% inhibition against both cholinesterases, whereas compounds 2, 8 and 11 showed 37, 42, 41% of inhibition towards BChE, being the most active. The highest metal-chelation capacity was observed with compound 8 (53.58 ± 2.06%). POM and DFT analyses are in good harmonization with experimental data.

In Vitro Antioxidant and Cytotoxic Activities of 18 Plants from the Erkowit Region, Eastern Sudan.

We investigated the antioxidant potential and cytotoxicity towards human CCRF-CEM leukemia cells of 57 extracts obtained from 18 plants collected in the Erkowit region, eastern Sudan. The antioxidant activity was determined by measuring the radical scavenging effects against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and N,N-dimethyl-p-phenylendiamine (DMPD), metal-chelation capacity, ferric-reducing (FRAP) and phosphomolibdenum-reducing antioxidant power (PRAP) methods using ELISA microtiter assays. Total phenol and flavonoid amounts of the extracts were determined spectrophotometrically. Cytotoxicity towards CCRF-CEM cells was evaluated by the resazurin reduction assay. Geranium favosum followed by Kalanchoe glaucescens, Malva parviflora, Aizoon canariense, and Coleus barbatus, respectively, possessed the highest antioxidant activity among the studied plants. Chrozophora oblongifolia and K. glaucescens exerted considerable cytotoxicity against CCRF-CEM leukemia cells. These plants may serve as source for the further development of natural antioxidant and antitumor agents.

Cholinesterase Inhibitory Activity of Some semi-Rigid Spiro Heterocycles: POM Analyses and Crystalline Structure of Pharmacophore Site.

BACKGROUND: Cholinesterase family consists of two sister enzymes; acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) which hydrolyze acetylcholine. Since deficit of acetylcholine has been evidenced in patients of Alzheimer's disease (AD), cholinesterase inhibitors are currently the most prescribed drugs for the treatment of AD. OBJECTIVE: our aim in this article was to investigate the inhibitory potential of five known compounds (2-6) with spiro skeleton against AChE and BChE using ELISA microplate assays. In addition to their ChE inhibitory effect, their physico-chemical properties were also calculated. Moreover, the present work aims at investigating the charge/geometrical effect of a hypothetical pharmacophore or bidentate site in a bioactive group, on the inhibition efficiency of spiro compounds 2-6 by using Petra/Osiris/ molinspiration (POM) and X-ray analyses. METHOD: In the present study, five compounds (2-6) with spiro skeleton have been synthesized and tested in vitro for their inhibitory potential against AChE and BChE using ELISA microtiter plate assays at 25 µg/mL. RESULTS: Results revealed that three of the spiro compounds tested exert more than 50% inhibition against one of cholinesterases. Compound 5 displayed 68.73 ± 4.73% of inhibition toward AChE, whereas compound 6 showed 56.17 ± 0.83% of inhibition toward BChE; these two previously synthesized compounds have been the most active hits. CONCLUSIONS: Our data obtained from screening of compounds 2-6 against the two cholinesterases indicate that three of these show good potential to selectively inhibit AChE or BChE. Spiro compounds 2, 5, and 6 exhibited the most potent activity of the series against AChE or BChE with inhibition values in the range 55-70%.

Cassia tora Linn.: A boon to Alzheimer's disease for its anti-amyloidogenic and cholinergic activities.

BACKGROUND: Drug discovery from natural products as alternatives for Alzheimer's disease (AD) is a current trend. For which plant is an alternative for searching potential molecule for treating AD. Availability of Cassia tora as weed and abundance in nature makes it as potential source. Many plants group under Leguminosae family has potential medicinal property of which Cassia tora is an appropriate choice, to know potency against AD. Etiology of AD is described by senile plaques and neurofibrillary tangles. The Aß42 has key major role in forming plaques by forming structures like protobirils, oligomers and final fibrilar like structures. Even at in vitro conditions, the peptides have a fibrilar like structure, which was exploited to preliminary screening of natural sources that may be effective in treating AD. HYPOTHESIS/PURPOSE: The design of the study was to unravel the potential medicinal property of Cassia tora for its antioxidant, cholinergic and aggregation inhibition activity. STUDY DESIGN: We evidenced that the methanol (MeOH), n-hexane (n-hex), petroleum ether (PE) and aqueous (aq) extracts from the leaves of Cassia tora (C. tora) were investigated for their inhibitory activity against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and anti-amyloidogenic assays. The antioxidant effect using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, total phenolic and flavonoid contents of the extracts were determined using Folin-Ciocaltaeu's and aluminum chloride (AlCl3) reagents, respectively. RESULTS: The methanol extract of C. tora exerted the highest inhibition against AChE (55.38 ± 2.28%) and BChE inhibition (50.02 ± 0.79%) at 100µg/ml concentration. The methanol extract was also found more active in the antioxidant test. The aggregation kinetics was monitored using thioflavin-T (ThT) assay and transmission electron microscopy (TEM) technique. CONCLUSION: The results showed that C. tora methanol extract is able to inhibit the Aß42 aggregation from monomers and oligomers and also able to dis-aggregate the pre-formed fibrils. The study provides an insight on finding new natural products for AD therapeutics.

Promising anticancer activity of Cyclotrichium niveum L. extracts through induction of both apoptosis and necrosis.

In this study, cytotoxic effects of the dichloromethane, ethyl acetate, ethanol, and aqueous extracts of the aerial parts of Cyclotrichium niveum (Boiss.) Manden. & Scheng (Lamiaceae) were evaluated. We tested HeLa, MCF-7 cancer cells, and MRC-5 and MCF-10A normal cells. The ethyl acetate and dichloromethane extracts induced cytotoxicity whereas the ethanol and aqueous extracts had no cytotoxic activity against both cancer cells. IC50 values of the dichloromethane extract were 353.0 ± 84.30 µg/ml, 114.8 ± 40.34 µg/ml, 39 ± 0.56 µg/ml, and 49 ± 13 µg/ml in HeLa, MCF-7, MRC-5, MCF-10A cells, respectively. IC50 values of the ethyl acetate extract were 117.0 ± 36.24 µg/ml in HeLa cells, 156.3 ± 19.86 µg/ml in MCF-7 cells, 1100 ± 340 µg/ml in MRC-5 cells and 7900 ± 1200 µg/ml in MCF-10A cells. Additionally, the ethyl acetate extract showed more selectivity to HeLa and MCF-7 cancer cells than MRC-5 and MCF-10A normal cells. Our results of HPLC analysis showed that apigenin in the ethyl acetate extract (2.2518 ± 0.0005 mg/100 mg extract) might be responsible of that selective cytotoxic effect. In the current work, we have shown for the first time that C. niveum has cytotoxic properties in the cancer cell lines tested.

Pteryxin - A promising butyrylcholinesterase-inhibiting coumarin derivative from Mutellina purpurea.

Pteryxin is a dihydropyranocoumarin derivative found in Apiaceae family. In this study, pteryxin, which was previously isolated from the fruits of Mutellina purpurea, was investigated for its inhibitory potential against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), which are the key enzymes in the pathology of Alzheimer's disease (AD). The compound was tested in vitro using ELISA microplate reader at 100 µg/ml and found to cause 9.30 ± 1.86% and 91.62 ± 1.53% inhibition against AChE and BChE, respectively. According to our results, pteryxin (IC50 = 12.96 ± 0.70 µg/ml) was found to be a more active inhibitor of BChE than galanthamine (IC50 = 22.16 ± 0.91 µg/ml; 81.93± 2.52% of inhibition at 100 µg/ml). Further study on pteryxin using molecular docking experiments revealed different possible binding modes with both polar and hydrophobic interactions inside the binding pocket of BChE. Top docking solution points out to the formation of two hydrogen bonds with the catalytic residues S198 and H438 of BChE as well as a strong π - π stacking with W231. Therefore, pteryxin as a natural coumarin seems to be a strong BChE inhibitor, which could be considered as a lead compound to develop novel BChE inhibitors for AD treatment.

Adonis sp., Convallaria sp., Strophanthus sp., Thevetia sp., and Leonurus sp. - Cardiotonic Plants with Known Traditional Use and a Few Preclinical and Clinical Studies.

Plants have been always a fruitful source of active metabolites against many human disorders, where cardiovascular (CV) diseases have been one of the major health problems all over the world. There are some clinically proved medicinal plants with cardioprotective effects such as Crataegus monogyna and C. oxyacantha (hawthorn) from Rosaceae. On the other hand, cardiac glycosides, present in a number of plant species, are wellknown for their cardiotonic activity. However, we encountered such a fact that very less number of studies are available on the source plants; e.g. Adonis vernalis, Convallaria majalis, Strophanthus kombe, Thevetia peruviana, Leonurus cardiaca, etc. Consequently, we herein aimed to cover all available data consisting of in vitro, in vivo, and human studies (if any) on cardiotonic effects of the aforementioned species through a wide literature search using Scopus, Web of Science as well as Pubmed.

HPTLC Fingerprinting and Cholinesterase Inhibitory and Metal-Chelating Capacity of Various Citrus Cultivars andOlea europaea.

Inhibitory activity of thirty-one ethanol extracts obtained from albedo, flavedo, seed and leaf parts of 17 cultivars of Citrus species from Turkey, the bark and leaves of Olea europaea L. from two locations (Turkey and Cyprus) as well as caffeic acid and hesperidin was tested against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), related to the pathogenesis of Alzheimer's disease, using ELISA microtiter assays at 500 µg/mL. Metal-chelating capacity of the extracts was also determined. BChE inhibitory effect of the Citrus sp. extracts was from (7.7±0.7) to (70.3±1.1) %, whereas they did not show any inhibition against AChE. Cholinesterase inhibitory activity of the leaf and bark ethanol extracts of O. europaea was very weak ((10.2±3.1) to (15.0±2.3) %). The extracts had either no or low metal-chelating capacity at 500 µg/mL. HPTLC fingerprinting of the extracts, which indicated a similar phytochemical pattern, was also done using the standards of caffeic acid and hesperidin with weak cholinesterase inhibition. Among the screened extracts, the albedo extract of C. limon 'Interdonato', the flavedo extracts of 'Kara Limon' and 'Cyprus' cultivars and the seed extract of C. maxima appear to be promising as natural BChE inhibitors.

Biological evaluation and docking studies of some benzoxazole derivatives as inhibitors of acetylcholinesterase and butyrylcholinesterase.

A series of 2,5-disubstituted-benzoxazole derivatives (1-13) were evaluated as possible inhibitors of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The results demonstrated that the compounds exhibited a broad spectrum of AChE and BChE inhibitory activity ranging between 6.80% and 90.21% except one compound which showed no activity against AChE at the specified molar concentration. Another derivative displayed a similar activity to that of reference drug (galanthamine) for inhibition of AChE and BChE. In addition, molecular docking of the compounds into active site of AChE was performed using recombinant human AChE (PDB ID: 4ey6) in order to understand ligand-protein interactions.

Anticholinesterase, antioxidant, analgesic and anti-inflammatory activity assessment of Xeranthemum annuum L. and isolation of two cyanogenic compounds.

CONTEXT: Xeranthemum annuum L. (Asteraceae) (XA) is an ornamental and medicinal species with limited bioactivity and phytochemical data. OBJECTIVE: Identification of anticholinesterase, antioxidant, anti-inflammatory and analgesic effects of the flower and root-stem (R-S) extracts of XA. MATERIALS AND METHODS: Anticholinesterase (at 100 µg mL-1) and antioxidant (at 1000 µg mL-1) effects of various extracts were evaluated via microtiter assays, while anti-inflammatory and analgesic effects of the R-S extracts were tested using carrageenan-induced hind paw oedema (100 and 200 mg kg-1) and p-benzoquinone (PBQ) writhing models (200 mg kg-1) in male Swiss albino mice. The R-S ethanol extract of XA was subjected to isolation studies using conventional chromatographic methods. RESULTS: Most of the extracts showed inhibition over 85% against butyrylcholinesterase and no inhibition towards acetylcholinesterase. The flower chloroform and the R-S ethyl acetate extracts were most effective (97.85 ± 0.94% and 96.89 ± 1.09%, respectively). The R-S ethanol extract displayed a remarkable scavenging activity against DPPH (77.33 ± 1.99%) and in FRAP assay, while the hexane extract of the R-S parts possessed the highest metal-chelating capacity (72.79 ± 0.33%). The chloroform extract of the R-S caused a significant analgesic effect (24.4%) in PBQ writhing model. No anti-inflammatory effect was observed. Isolation of zierin and zierin xyloside, which were inactive in anticholinesterase assays, was achieved from the R-S ethanol extract. DISCUSSION AND CONCLUSION: This is the first report of anticholinesterase, antioxidant, analgesic and anti-inflammatory activities and isolation of zierin and zierin xyloside from XA. Therefore, XA seems to contain antioxidant and BChE-inhibiting compounds.

Tyrosinase and Cholinesterase Inhibitory Potential and Flavonoid Characterization of Viola odorata L. (Sweet Violet).

Inhibitory potential of the dichloromethane, ethyl acetate, ethanol, and aqueous extracts of Viola odorata L. (VO) was investigated against tyrosinase (TYR) and cholinesterases by microplate assays. The antioxidant activity was tested using six in vitro assays. Only the ethanol extract inhibited TYR (80.23 ± 0.87% at 100 µg mL-1 ), whereas none of them were able to inhibit cholinesterases. The extracts were more able to scavenge NO radical (31.98 ± 0.53-56.68 ± 1.10%) than other radicals tested, and displayed low to moderate activity in the rest of the assays. HPLC analysis revealed that the aqueous extract of VO contained a substantial amount of vitexin (18.81 ± 0.047 mg g-1 extract), while the ethanol extract also possessed rutin (1.31 ± 0.013 mg g-1 extract) and vitexin (4.65 ± 0.103 mg g-1 extract). Furthermore, three flavonoids (rutin, isovitexin, and kaempferol-6-glucoside) were isolated from the ethanol extract. This is the first report on TYR inhibitory activity of VO as well as presence of vitexin and isovitexin in this species. Copyright © 2015 John Wiley & Sons, Ltd.

In vitro cholinesterase inhibitory and antioxidant effect of selected coniferous tree species.

OBJECTIVE: To explore cholinesterase inhibitory and antioxidant effect of six coniferous trees (Abies bornmulleriana, Picea pungens, Juniperus communis, Cedrus libani, Taxus baccata, and Cupressus sempervirens var. horizantalis). METHODS: Acetone (Ace), ethyl acetate (EtOAc), and ethanol (EtOH) extracts prepared from the needles and shoots of the six coniferous trees were screened for their acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity at 100 µg/mL. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and N,N-dimethyl-p-phenylendiamine (DMPD) radical scavenging, metal-chelation capacity, ferric-(FRAP) and phosphomolibdenum-reducing antioxidant power (PRAP) assays. All of the assays were performed in ELISA microplate reader. Total phenol and flavonoid amounts in the extracts were determined spectrophotometrically. RESULTS: Among thirty-six extracts in total, the shoot-Ace extract of Cupressus sempervirens var. horizantalis exerted the highest inhibition against AChE [(54.84±2.51)%], while the needle-Ace extract of Cedrus libani was the most effective in inhibiting BChE [(67.54±0.30)%]. The highest DPPH radical scavenging effect, FRAP and PRAP was observed in the shoot-Ace and EtOAc extracts from Taxus baccata, whereas all the extracts showed a variable degree of scavenging effect against DPMD radical. The shoot-EtOAc extract of Cedrus libani had the highest metal-chelation capacity [(58.04±0.70)%]. The shoot extracts of Taxus baccata were determined to have the richest total phenol content, which may contribute to its marked antioxidant activity. CONCLUSIONS: The conifer species screened in this study may contain cholinesterase-inhibiting and antioxidant properties, which might be useful against Alzheimer's disease.

Comparative antioxidant activity appraisal of traditional Sudanese kisra prepared from two sorghum cultivars.

The effect of fermentation and heating on the antioxidant activity of the fermented and fermented baked (kisra) dough prepared from two Sorghum cultivars (Tabat and Wad Ahmed) was evaluated. Kisra prepared from Tabat cultivar showed higher DPPH (1,1-diphenyl-2-picrylhydrazyl radical) scavenging and ferric reducing/antioxidant power (FRAP) than that of the Wad Ahmed cultivar. Baking improves the DPPH and FRAP of the kisra prepared from two cultivars. Baking caused a variable effect on the total phenol, tannins and flavonoids content across different periods of fermentation where an increase was mainly observed for samples subjected to longer periods of fermentation. A positive high correlation between the total phenol and antioxidant activity, using the DPPH and FRAP assays, was obtained for kisra prepared from both cultivars. The same observation was obtained for tannin content. In conclusion, fermentation and heating improve the antioxidant capacity of the sorghum grains from Tabat and Wad Ahmed cultivars.

Synthesis and different biological activities of novel benzoxazoles.

A series of 2-[4-(4-substitutedbenzamido/phenylacetamido/butanamido)phenyl]-5-ethylsulphonyl-benzoxazole derivatives were synthesized and biologically evaluated as possible antimicrobial agents and inhibitors of tyrosinase, acetylcholinesterase (AChE), and butyrylcholinesterase (BChE). The results demonstrated that the synthesized compounds exhibited a broad spectrum of activity with minimum inhibitory concentration (MIC) values of 128-16 µg/ml against some Gram-positive, Gram-negative bacteria as well as Candida albicans and C. krusei. The compound 10 displayed higher activity in this series against methicilline resistant Staphylococcus aureus (MRSA) with a MIC value of 16 µg/ml than the compared control drugs ampicillin and ceftriaxone. Compound 14 showed moderate tyrosinase inhibition, however, none of the compounds showed effect as inhibitor of AChE and BChE.

An in vitro perspective to cholinesterase inhibitory and antioxidant activity of five Gentiana species and Gentianella caucasea.

The dichloromethane (DCM), ethyl acetate (EtOAc), and methanol extracts from the leaves, roots, and flowers of the five species of Gentiana (Gentiana asclepiadea, Gentiana cruciata, Gentiana olivieri, Gentiana septemfida, and Gentiana verna) and Gentianella caucasea were investigated for their inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) and antioxidant effect using 2,2-diphenyl-1-picrylhydrazyl radical scavenging, metal-chelation capacity, and ferric-reducing antioxidant power assays. Total phenol and flavonoid contents of the extracts were determined spectrophotometrically. The presence of some characteristic compounds found in Gentiana species (gentiopicroside, swertiamarin, isoorientin, isovitexin and vitexin) was analyzed in the extracts by thin layer chromatography. The flower DCM extract of G. verna exerted the highest inhibition against AChE (53.65 ± 1.03%), whereas the root EtOAc extract of G. cruciata was the most effective in BChE inhibition assay (50.72 ± 0.75%) at 100 µg ml⁻¹. The extracts of G. verna were also found to be more active in the antioxidant tests.

Profiling of in vitro neurobiological effects and phenolic acids of selected endemic Salvia species.

The ethyl acetate and methanol extracts from 16 Salvia L. species were screened for their inhibitory activity against acetylcholinesterase, butyrylcholinesterase, lipoxygenase, and tyrosinase; the enzymes linked to neurodegeneration. Their antioxidant activity was also tested using DPPH radical scavenging, metal-chelation, and ferric-reducing antioxidant power (FRAP) assays. Total flavonoid content of the extracts was determined by AlCl3 reagent, while HPLC technique was applied for analysis of various phenolic acids in the extracts. The extracts exerted weak cholinesterase and tyrosinase inhibition, and remarkable inhibition against lipoxygenase (13.07±2.73-74.21±5.61%) at 100µgml-1. The methanol extracts showed higher antioxidant activity in DPPH radical scavenging and FRAP assays. The extracts were analyzed for their gallic, protocateuchic, p-hydroxy-benzoic, vanillic, caffeic, chlorogenic, syringic, o- and p-coumaric, ferulic, rosmarinic, and tr-cinnamic acid contents and the methanol extract of Salvia ekimiana (153.50mg100g-1) was revealed to be the richest in terms of rosmarinic acid.

Inhibitory potential of the leaves and berries of Myrtus communis L. (myrtle) against enzymes linked to neurodegenerative diseases and their antioxidant actions.

The dichloromethane (DCM), acetone, ethyl acetate and methanol extracts of the leaves and berries of Myrtus communis L. were screened against acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and tyrosinase (TYRO), the enzymes linked to neurodegenerative diseases, at 200 µg ml⁻¹. Antioxidant activity was tested using radical scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and N,N- dimethyl-p-phenylenediamine (DMPD) radicals, metal chelation capacity, ferric-reducing antioxidant power (FRAP) and phosphomolybdenum-reducing antioxidant power (PRAP) assays. Total phenol and flavonoid quantification of the extracts was calculated spectrophotometrically. The extracts showed a moderate AChE (17.49 ± 3.99% to 43.15 ± 1.55%) and TYRO inhibition (4.48 ± 1.50% to 40.53 ± 0.47%). The leaf extracts were ineffective against BChE, while the berry extracts displayed inhibition between 21.83 ± 3.82% and 36.80 ± 2.00%. The polar extracts exerted remarkable scavenging effect against DPPH and DMPD and also in the FRAP assay, where the DCM extract of the berries had the best metal chelation capacity (79.29 ± 1.14%). This is the first study that demonstrates in vitro neuroprotective effects of myrtle.

Assessment of antimicrobial, insecticidal and genotoxic effects of Melia azedarach L. (chinaberry) naturalized in Anatolia.

In this study, antibacterial, antifungal, insecticidal and genotoxic activities of the fruit and leaf extracts of Melia azedarach of Turkish origin were evaluated for the first time. The antimicrobial activity was assessed against Gram (+) and (-) bacteria, four Candida species and three dermatophytic fungus (Trichophyton rubrum, Epidermophyton floccosum and Microsporum gypseum). The insecticidal activity of the methanolic fruit extract was performed against the larvae of Aedes aegpyti, Culex pipiens and Culex quinquefasciatus. The genotoxicity of this extract was evaluated against Drosophila melanogaster by somatic mutation and recombination test. The extracts showed higher antibacterial effect against Gram (-) strains (16-32 µg ml(-1) of minimal inhibition concentration, MIC), while the leaf extracts were more effective on Candida albicans (32 µg ml(-1)of MIC). The extracts did not exhibit insecticidal activity and genotoxicity. Total phenol and flavonoid contents of the extracts were determined spectrophotometrically, and the ethyl acetate extract of the leaves was the richest in total flavonoids.

Evaluation of cholinesterase inhibitory and antioxidant activities of wild and cultivated samples of sage (Salvia fruticosa) by activity-guided fractionation.

In European folk medicine, Salvia species have traditionally been used to enhance memory. In our previous study of 55 Salvia taxa, we explored significant anticholinesterase activity of cultivated S. fruticosa. In this study, we compared the inhibitory activity of dichloromethane, ethyl acetate, and ethanol extracts of 3 wild-grown samples and 1 cultivated sample of S. fruticosa against acetylcholinesterase and butyrylcholinesterase enzymes (which are associated with pathogenesis of Alzheimer's disease) by using the spectrophotometric Ellman method. Antioxidant activities were assessed by determining 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity, iron-chelating capacity, and ferric-reducing antioxidant power. The dichloromethane extract of the cultivated sample was then subjected to fractionation by using open column chromatography and medium-pressure liquid chromatography to obtain the most active fraction by activity-guided fractionation. All fractions and subfractions were tested in the same manner, and inactive subfractions were discarded. The essential oil of the cultivated sample was analyzed by gas chromatography-mass spectrometry.