DCM Extracts Research Articles

HPLC-UV-ESI-MS phyto-analysis and biological activities of Rhanterium adpressum extracts (Asteraceae) from southern Algeria

Biological activities of dichloromethane, ethyl acetate and 1-butanolextracts from Rhanterium adpressum flowers was evaluated. The antioxidant activities were measured by three different methods, namely DPPH, phosphomolybdenum and ABTS assays. The results suggested that all extracts possessed an antioxidant activity and capacity. In addition, the dichloromethane extract of April showed the best antimicrobial activity against Pseudomonas aeruginosa (MIC = 6.5 mg/ml) and Staphylococcus aureus (MIC = 20mg/ml). However, anti-diabetic activity tested using an in vitro model by α-glucosidase and α-amylase inhibition testsshowed that the extracts was found to be more effective in inhibiting the activities of maltase with an IC 50 values ranging from 125 to 397 µg/ml when compared to that of α-amylase with IC 50 varying from 1040 to 2710 µg/ml. The phenolic analysis of the most active extract obtained from Rhanterium adpressum has been done by HPLC-UV-ESI/MS. Several flavonoids and phenolics were identified, such as syringic and vanillic acids which were known as important antioxidant phenols.The obtained results suggest that plant extracts could be used as an ingredient in treatment of infectious diseases or as antioxidant. • Antioxidant activity was assessed by three tests in vitro . All extracts exhibited a high antioxidant capacity compared with vitamin C. • Antibacterial activity indicated that DCM extract has been considered as the best extract (MIC = 6.5mg/ml). • Anti-diabetic activity tested using an in vitro model by α-glucosidase and α-amylase inhibition showed a higher inhibition i.e. lower IC 50 (201.09±0.08 µg/mL; 185±0.02 µg/ml & 171±0.01µg/ml for DCM, EtAc & BuOH respectively). • HPLC-UV-ESI/MS analysis of the most active extracts shows several important bioactive compounds (vanillic acid, synringic acid, quercetine).

GC/TOF-MS-based phytochemical analysis, in vitro antiproliferative effects, antioxidant and antibacterial activity of Sarcophyte sanguinea subsp. piriei (Hutch.) B. Hansen

Antibiotic resistance necessitates the need for continued search for new antimicrobial compounds. In this study, we report the antibacterial activity, cytotoxic, and antioxidant activity of Sarcophyte sanguinea subsp. piriei (Hutch.) B. The extracts were evaluated for antibacterial activity using the agar well diffusion, micro-dilution, minimum bactericidal concentrations (MBC) and lactate dehydrogenase (LDH) release assay. Furthermore, the cytotoxic effects of extracts were determined using MTT assay against Human Embryonic Kidney (HEK293), Human Breast Endocrine Cells (SKBR-3), Human Colorectal Carcinoma Cells (Caco-2) and Human Hepatocellular Carcinoma (HepG2) cells. The extracts were also assessed for antioxidant activity against 2,2-diphenyl-1-picryl hydrazyl (DPPH), 2–2′-Azino-di-[3-ethylbenzthiazoline sulfonate (ABTS), nitric oxide radicals and Fe2+ chelating assays. The methanol extract (MeOH) was subjected to GC-TOF-MS analysis. MeOH extract exhibited highest zone of inhibition (ZI) of 15.67 mm against Staphylococcus aureus. The extract further exhibited 67.0% inhibition against Escherichia coli in a lactate dehydrogenase (LDH) membrane damage assay. In the cytotoxicity assay, MeOH and DCM extracts had LC50 value of 237 µg/ml and 221 µg/ml against SKBR-3 cell line respectively. Gas-chromatography time-of-flight mass spectrometry (GC/TOF-MS) analysis of methanol extract indicated the presence of 5-aminoimidazole-4-carboxamide-1-ád-ribofuranosyl 5′-monophosphate (36.827%), 3-O-methyl-d-glucose (36.827%) and phthalic acid, di(oct‑3-yl) ester (8.161%). Even though the investigated plant is suggested to have anticancer activities by traditional healers, the extracts had little inhibition on selected cancerous cell lines. Given the good antibacterial and antioxidant activity of the extracts, the plant may act as an immune booster and prevent infections in immunosuppressed cancer patients. Detected compounds may be influence the observed biological activity of the plant.

Expression of the Fab enzymes (Fab I and Fab Z) from Plasmodium falciparum after exposure to Artemisia afra plant extracts and drugs screening.

The emergence and spread of drug resistance of the malaria parasite to the main treatment emphasize the need to develop new antimalarial drugs. In this context, the fatty acid biosynthesis (FAS_II) pathway of the malaria parasite is one of the ideal targets due to its crucial role in parasite survival. In this study, we report the expression and the affinity binding of Fab_I and Fab_Z after exposure to the parasite with different extracts of the Artemisia afra. The parasites were exposed for 2days to different extracts. Gene expression was done to determine the level of expression of the fab enzymes after treatments. A GCMS was run to determine the different compounds of the plant extracts, followed by a virtual screening between the fab enzymes and the active compounds using Pyrex. The results showed different expression patterns of the Fab enzymes. Fab_I expression was downregulated in the W2 and D6 strains by the ethanolic extract but was increased by Hexane and DCM extracts. A different expression pattern was observed for Fab_Z. It was all upregulated except in the D6 strain when exposed to the ethanolic and hexane extracts. Virtual screening showed an affinity with many compounds. Hits compounds with high binding energy were detected. 11alphaHydroxyprogesterone and Aspidospermidin-17-ol were found to have high binding energy with Fab_I respectively (-10.7kcal/mol; -10.2kcal/mol). Fab_Z shows also high affinity with 11alpha-Hydroxyprogesterone (-10kcal/mol) and Thiourea (-8.4kcal/mol). This study shows the potential of A. afra to be used as a new source of novel antimalarial compounds.

Evaluation of Bioactive Secondary Metabolites from Ponyfish Associated Bacteria (<i>Photobacterium leiognathi</i>)

The marine environment continues to surprise us by producing novel bioactive substances with a wide range of benefits for humans. Materials and Methods: Marine bioluminescent bacteria Photobacterium leiognathi was isolated from pony fish, Secutorruconius which was confirmed with microscopic and molecular characterization. The secondary metabolite of the isolated bacteria was extracted with dichloromethane. The chemical fingerprinting of the isolated metabolite was analyzed through TLC, FT-IR, and HPLC. The nature of the compound present in the metabolite was identified in the gas chromatography-mass spectrometry analysis (GC - MS). The isolated extract was investigated for its antibacterial property against 10 human pathogenic bacteria and also its antioxidant activity using different assays such as 1, 1-Diphenyl-2-picrylhydrazyl, Phosphomolybdenum, Metal chelating, Hydroxyl radical scavenging and hydrogen peroxide scavenging activity. Results: The Presence of functional groups including phenols, sugars, and amino acids in the extracts were identified by TLC. Totally, nine peaks were obtained for the crude extract through the FTIR spectrum range of 400 to 4000 cm-1 for the active sample. The DCM extract showed a broad spectrum of antibacterial activity against the six human bacterial pathogens. Secondary metabolites from the bioluminescent bacteria, P. leiognathi, have strong antioxidant properties. These results will be instrumental in developing novel products with biosensors and bio-imaging applications using P. leiognathi.

The Effect of Acidic Ternary Deep Eutectic Solvent Treatment on Native Lignin

Ternary deep eutectic solvents (DESs) are gaining increased attention to serve as a cheap green alternative medium for the processing of lignocellulosic biomass. For example, mixtures of choline chloride (ChCl), ethylene glycol (EG), and oxalic acid (OA) were recently explored for the fractionation of lignocellulosic biomass into its main components. Interestingly, during this processing, the recovered lignin was structurally modified by incorporation of EG, which altered its solubility properties and led to the need for different lignin recovery strategies. This offers an excellent starting point for a deeper investigation of the effect of acidic DES systems on the structure of lignin. In particular, native-like residual enzyme lignins (RELs) that are hard to completely dissolve in organic solvents are specifically suitable for this task. Here, a ternary DES is used consisting of ChCl/EG with OA or trifluoromethanesulfonic acid (HOTf) as a third component. The results showed that both solvent systems led to high EG incorporation into REL. The HOTf system showed a lesser extent of lignin depolymerization at similar modification levels as it already induced modification at lower temperature (25–30 °C). Low recovery yields from typical acidic precipitation were observed for treatment with both acidic DES systems. Analysis of THF and DCM extracts showed that the products in the water phase included small EG modified lignin fragments and aromatic monomers released from lignin aryl ether linkage cleavage. This analysis details the types of other products that can be expected and where these will end up during fractionation. These results show that the treatment of lignin with acidic DES in the presence of alcohols leads to low- and high-molecular-weight products that are not effectively recovered by typical precipitation procedures.

Genotoxic hazard and oxidative stress induced by wastewater irrigated soil with special reference to pesticides and heavy metal pollution

Due to enhancement of industrial growth and urbanization, soil contamination is increasing prominently. Therefore, it is important to examine possible adverse effects of industrial waste. Soil samples were might to be polluted with several heavy-metals and pesticides. Gas chromatographic results showed occurrence of high-level of organochlorine and organophosphate pesticides in studied soil samples. Genotoxicity of soil extracts was assessed using environmental-risk assessment models. Soil samples were extracted in hexane and dichloromethane solvents and were evaluated for genotoxic potential by prokaryotic (Ames test, plasmid nicking assay and E. coli K-12 DNA repair defective mutants) and eukaryotic (Allium cepa root chromosomal aberration and Vigna radiata seed-germination test) bioassays. Strain TA98 was found the most susceptible among soil extracts. The mutagenicity of hexane soil extract from wastewater irrigation was found to be higher than that of DCM samples in terms of mutagenic index, mutagenic potential, and induction factor for Ames strains. The damage in DNA repair defective mutants of hexane extracts were found higher compared to DCM extracts at dose of 20 μl/ml of culture. Survival in polA, lexA and recA mutants were 39%, 47% and 55% while treated with hexane extract. Allium cepa test, mitotic index was decreased in dose-dependent way and various kinds of chromosomal aberrations were found. Vigna radiata seeds germination and other parameters were also affected when treated with wastewater irrigated (WWI) soil. Oxidative stress in V. radiata roots were also showed under CLS microscope. Genotoxicity of WWI soil extract was also confirmed by plasmid nicking test. Our study provides possible explanation for the assessment of potential health and environmental hazards of the industrial region.

In vitro antiplasmodial activity and cytotoxicity of extracts and chromatographic fractions of twigs from Pappea capensis EckI & Zeyh. (Sapindaceae)

Ethnopharmacological relevanceThe Vha-Venda people of South Africa use Pappea capensis EckI & Zeyh. (Sapindaceae) twigs to treat malaria and its related symptoms. Aim of studyThe main aim of this study was to evaluate the antiplasmodial and cytotoxic activity of P. capensis extracts and chromatographic fractions. Spectroscopy analysis was conducted using 1H NMR and GC-MS to tentatively identify the major classes of compounds and phytoconstituents that can be attributed to the observed antiplasmodial bioactivity. Materials and methodsPappea capensis twigs were dried and then ground to fine powder. A solvent mixture of dichloromethane: methanol: water (1:0.5:0.5, v/v) was used to extract. The polar extract was separated from the non-polar. The organic extract was dried to yield a DCM (I = 60 g) extract. The methanol in the aqueous extract was evaporated using a rotary vapour and the remaining water freeze dried to yield a water extract (II = 287 g). Extract I was further partitioned using a solvent mixture of DCM: MeOH (1:1, v/v), separated and concentrated under vacuum to yield dichloromethane (III = 40 g) and methanol (IV = 15 g) extracts. A water-based decoction (V = 10 g) was also prepared to establish the clinical relevance of the preparation administered by Vha-Venda people in South Africa. Extracts II, III and IV were further subjected to silica column chromatography, eluting with a series of different solvents with increasing polarity to yield a total of 25 fractions (A – Y). In vitro antiplasmodial tests on Plasmodium falciparum (NF54) and cytotoxicity screens on mammalian L-6 rat skeletal myoblast cells were performed on all extracts and fractions. Selectivity indices (SI) were also computed for all tested extracts and fractions which were further subjected to 1H NMR spectroscopy and GC-MS analysis for the identification of the major classes of compounds present in the extracts. ResultsFrom the assayed extracts, only extract I (IC50 = 2.93 μg/ml; SI = 14), III (IC50 = 2.59 μg/ml; SI = 21) and IV (IC50 = 3.56 μg/ml; SI = 13) demonstrated the best antiplasmodial activity and selectivity. Of all assayed fractions, only N (0.6 μg/ml; SI = 91), D (0.85 μg/ml; SI = 37) and E (0.91 μg/ml; SI = 30) depicted the best antiplasmodial activity and selectivity. The 1H NMR analysis of the extracts and fractions identified the prominent class of constituents to be aliphatic based which was tentatively identified as terpenoids. When further GC-MS analysis was conducted, the presence of lupin-3-one, lupeol acetate, α-amyrin, and β-amyrin phytoconstituents were tentatively confirmed. These constituents are triterpenoids with established antiplasmodial activity which can be tentatively attributed to the bioactivity observed in P. capensis twigs. ConclusionThe study validates the ethnomedicinal use of P. capensis for malaria treatment. It demonstrated the potential of discovering novel antiplasmodial constituents that could serve as drug hits through dereplication approaches where known compounds with established antimalarial activity can be bypassed to focus on the unknown.

Characterization of Essential Oil, Estimation of Phenolic and Flavonod Content and Biological Activities of Ephedra pachyclada BOISS

The northern areas of Nepal are rich in biodiversity and contain a large number of medicinal plant species including the Genus Ephedra of evergreen gymnosperm, belonging to the family Ephedraceae. The plants have been used by the peoples of the Himalayan region for the treatment of asthma, blood pressure, and gastritis for many years. This study was aimed for the evaluation of phytochemical, antioxidant, and antimicrobial activities of methanol, ethyl acetate, dichloromethane, and n-hexane extracts, and gas chromatography-mass spectrometry (GCMS) profiling of the essential oil of the aerial parts of Ephedra pachyclada Boiss from Mustang district of Nepal. Antioxidant activity was evaluated by 2,2-dipheny-1-picrylhydrazyl (DPPH) free radical scavenging assay and antimicrobial activity by the agar well diffusion method. Total phenolics and total flavonoid content were determined by Folin-Ciocalteu and aluminum chloride colorimetric method, respectively. The methanol extract contained the highest total phenolic content of 54.42 ± 1.40 mg GAE/g followed by the ethyl acetate (46.84 ± 0.62 mg GAE/g), DCM extract (19.58 ± 0.24 mg GAE/g), and the lowest TPC was shown for n-hexane extract (5.21 ± 1.40 mg GAE/g) of the dry weight. The methanol extract showed the maximum TFC of 33.28 ± 0.48 mg QE/g, followed by ethyl acetate extract (31.73 ± 0.52 mg QE/g), DCM extract (31.64 ± 0.56 mg QE/g), and the least value was obtained for the n-hexane extract (21.44 ± 2.91 mg QE/g). The methanol extract showed the highest antioxidant activity with a half-maximal inhibitory concentration (IC50) of 37.81 ± 2.24 μg/mL. The methanol extract showed potent activity against Escherichia coli (ATCC 25922), and Staphylococcus aureus (ATCC 25923) with zones of inhibition of 13 mm and 12 mm respectively. Ethyl acetate extract showed a slight potency against Staphylococcus aureus (ATCC 25923) with a zone of inhibition of 9 mm. The essential oil contained diisooctyl phthalate (46.90%), dodecane, 2,6,11-trimethyl-(16.35%), dodecane, 4,6-dimethyl- (11.59%), tetrapentacontane (11.56%), and myrtenol (4.37) as the major compounds. The plant exhibited significant antioxidant and antimicrobial activities which could be used as the source to isolate the active natural antioxidant and antimicrobial agent as the drug candidate in the future drug discovery process.

Bioassay-guided Isolation of Triterpene Compounds from Dillenia suffruticosa and Their Cytotoxic Activities Against Cancer Cells

Previous studies showed the ability of Dillenia suffruticosa to inhibit the growth of cancer cells by means of cell cycle arrest and apoptosis, thus validating the traditional use of the plant in treating cancer. Therefore, the present study was designed to isolate and elucidate the bioactive compounds responsible for the anticancer properties of D. suffruticosa extract. Bioassay-guided fractionation of the most potent fractions of DCM and EtOAc extract of D. suffruticosa was performed via column chromatography followed by purification using preparative HPLC. The structures of the isolated compounds were elucidated by NMR spectroscopy. Biological experiment by MTT assay and a series of column chromatography resulted in the isolation of three triterpene compounds. However, only the structure of compound (3) was confirmed as 1-isopropenyl-4α, 4β, 8, 10, 14-pentamethyl-icosahydro-cyclopenta[a]chrysene-3, 17-diol. Another two compounds were labelled as (1) and (2), which currently are unidentified due to unsuccessfulness in the full analysis of the spectroscopic data that enable the assignment of all protons, carbons, and confirmation of the structure. Compound (2) isolated from DCM extract of D. suffruticosa was most cytotoxic towards the selected cancer cells compared to the other two compounds and selected chemotherapeutic drugs, tamoxifen, and cisplatin. The ability of the isolated compounds to inhibit the growth of cancer cells indicates that these compounds are the bioactive constituents in D. suffruticosa that is mainly responsible for the cytotoxic activities. For this reason, these isolated compounds could be used as a means for the standardisation of herbal product from D. suffruticosa.

Extraction, Isolation, and Identification of Flavonoid from Cassia Singueana Aerial Parts.

The flavonoids contained in C. singueana aerial parts were extracted, identified and characterized. Sequential Soxhlet extracts were subjected to preliminary phytochemical screening and flavonoid quantification. The results showed that maximum yield of the flavonoid (8.435 mg/g) were obtained from DCM extract. The DCM extract was subjected to flash chromatography for isolation of flavonoid. Isolated flavonoid was characterized using various spectroscopic techniques. On the basis of chemical and spectral analyses structure was elucidated as 2-3, (4-dihydroxyphenyl)-3, 5, 7-trihydroxy-4H-chromen-4-one, a flavonoid. Keywords: Cassia Singueana, Flavonoids, Flash chromatography, Quercetin

Chemical Composition of the Essential Oil and the Solvent Extracts of Curcuma Pseudomontana Rhizomes and in Vitro Evaluation of their Antioxidant and Anti-Inflammatory Activities

The phytoconstituents of the essential oil isolated by hydrodistillation and solvent extracts (PE, BZ, DCM, EtOAc and EtOH) of mature rhizomes of turmeric (Curcuma pseudomontana) were analyzed by GC–MS. The major constituents identified in the oil and the extracts were Benzofuran (6.93%), γ-Curcumene (11.18%), β-Elemenone (22.14%), Germacrone (15.15%), Phenol, 2-Methyl-5-(1,2,2-trimethylcyclopentyl)- (29.58%), Germacrene (8.78%), Italicene (15.30%), Xanthorrhizol (56.52%), Pseudocumenol (24.56%) and Epicurzerenolne (4.95%). Analyzing the anti-inflammatory activity of essential oils (EO), and solvent extracts using HRBC stabilization and antioxidant activity. The results of GC MS analysis showed that while Pseudocumenol the major componenets in the essential oil. Xanthorrhizol is the major component in BZ, DCM and EtOH extracts of the matured rhizomes of C. psedomonata. The results of EO and the solvent extracts from Curcuma possessed good anti-inflammatory and antioxidant activities and hence can be used in natural preservative ingredients in food and/ or pharmaceutical industries.

Anti-Hepatitis C Virus Activity of Various Indonesian Plants from Balikpapan Botanical Garden, East Borneo

Background: Hepatitis C Virus infection is a serious health problem that leads to chronic liver disease, liver cirrhosis, hepatocellular carcinoma, which causes high morbidity. Direct-Acting Antiviral Agents have been used as anti-hepatitis C Virus therapy. However, it was covered only in limited patients due to the high cost. Moreover, serious side effects and resistance cases were also reported in some HCV genotypes. Objective: This research aimed to find new anti-HCV from some Indonesia plants collected from Balikpapan Botanical Garden, East Borneo. Methods: Twenty-one leaf and stem barks extracts were successively extracted in n-hexane, dichloromethane, and methanol. Extracts were screened for their anti-HCV activity under in vitro culture cells in the concentration of 30 µg/mL. Plant extracts were inoculated in the Human Hepatocellular 7it and infected with HCV Japanese Fulminant Hepatitis strain 1a. Determination of 50% Inhibitory Concentration (IC50) value was further conducted at concentration of 100; 30; 10; 1; 0.1; 0.01 µg/ml of extracts. Results: In vitro anti-HCV activity revealed that among 21 plants extract, 11 extracts, namely, n-hexane extract from Luvunga scandens leaves, DCM extract from the leaf of L. scandens, Artocarpus sericicarpus, Artocarpus dadah, Eusideroxylon zwageri, Neolitsea cassiaefolia, methanol extract from A. sericicarpus and A. anisophyllus leaves, DCM extract from A. anisophyllus and A. elmeri stem bark, methanol extract from A. dadah stem bark, having potential inhibition with IC50 range 0.08 ± 0.05 to 12.01 ± 0.95 µg/mL. Conclusions: These results indicate that the eleven extracts could be good candidates as sources of anti-HCV agents.

Chemical composition and antioxidant activity of extracts from Moroccan fresh fava beans pods (Vicia Faba L.).

In Morocco, fava beans are widely used as a main meal or as an ingredient in various traditional recipes, in the form of fresh ripe seeds or dry seeds. In the past, the tender skin of bean pods was also used in certain specific dishes, thus diversifying the diet. However, the peels of the tender bean pods are currently less or not used and considered waste. In Moroccan, fava bean pods peels, traditionally used in food in the past, are today considered as waste. The valorization of fresh fava bean pods could revitalize the use of the specific dishes and diversify the diet. For this reason, the research aimed to assess the nutritional values and biological compounds of the whole fresh fava bean pods (Vicia faba L.). Evaluate the content of nutrients, total phenolic, flavonoids and tannin contents and antioxidant activity in different extracts of the tender pods of the fava bean (Vicia faba L.). The proximate composition and minerals were determined using AOAC methods. The total phenolic compounds by the Folin-Ciocalteu reagent, the total flavonoids were analyzed using aluminum chloride colorimetric method, the tannins by method of vanillin in an acidic medium and the antioxidant activity was evaluated by DPPH method. The results show that the fresh fava bean pods have a moisture content of 87.31 ± 0.25%, ash 4.67 ± 1.03, and protein 29.11 ± 3.20 g/100 g. The legume samples also contain potassium (1946.8±4.61), phosphorus (483.8 ± 3.14), and calcium (399.6 ±2.25) mg/100 g of dry matter representing at last 40-50% of the RDI. The content of the different extracts of (Vicia faba L) varied from 49.5 to 594.4 mg GAE/ g for the total phenols, from 0.7 mg to 3.4 mg QE/g for flavonoids, and from 4.9 mg to 73.91 mg TAE/g dry weight for tannins. The evaluation of the antioxidant activity in the various extracts revealed a better activity in the methanolic extract (IC50=491.2 μg/mL) compared to others extracts: the MeOH/water extract (IC50=606.61 μg/mL), DCM/ MeOH extract (IC50 = 642.67 μg/mL) and DCM extract below of 50%. This study shows that fava bean pods, traditionally used in food, are rich in macro and micronutrients and bioactive substances, which demonstrates their potential contribution to human food and nutritional security.

Chemical Constituents of Phyllanthus acidus (L.) Skeels

Abstract: Background: This study describes the isolation of chemical constituents from the leaves and bark of Phyllanthus acidus, a medicinal plant widely used for the treatment of several ailments. Materials and Methods: The crude extract of P. acidus was chromatographed on a gravity column dry packed with silica gel and was fractionated by increasing proportions of acetone in CH2Cl2. The purified isolates were subjected to NMR for structure elucidation. Their structures were identified mainly by using 1H-NMR and by comparing current NMR data with those reported in the literature. Results: The results showed that the air-dried leaves extracts of the plant afforded mixture of β-sitosterol and stigmasterol while the bark yielded mixture of phyllanthol, α-amyrin, β-amyrin and lupeol. Conclusion: The air-dried of DCM extract of P. acidus resulted in the isolation of the mixture of phyllanthol, α- amyrin, β-amyrin and lupeol from the bark and mixture of β-sitosterol and stigmasterol from the leaves of the plant. It is important to conduct detailed pharmacological and toxicity investigations as well as clinical studies using different doses or concentrations to verify its traditional uses. Key words: Phyllanthus acidus, Karamay, Phyllanthol, Lupeol, Amyrin, Sitosterol, Stigmasterol.

Cytotoxic and Apoptotic Effect of Iris taochia Plant Extracts on Human Breast Cancer (MCF-7) Cells

Introduction: Iris taochia is an endemic plant in Turkey. Iris species has many biological effects such as antibacterial, antiinflammatory, antioxidant and anticancer properties. Apoptosis is a programmed cell death and this mechanism regulates the death of cancer cells. Purpose: The aim of our work is to investigate how the Iris taochia extracts affect the apoptotic activity in the MCF7 cells. Methods: Cytotoxic dose and cell viability is determined by the MTT assay. Bad, Bax, Bcl-2, Bcl- W, Bid, Bim, Caspase 3, Caspase 8, CD40, CD40L, cIAP-2, CytoC, DR6, Fas, FasL, HSP27, HSP60, HSP70, HTRA, IGF-I, IGF-II, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IGFBP-5, IGFBP-6, IGF-1sR, Livin, p21, p27, p53, SMAC, Survivin, sTNF-R1, sTNF-R2, TNF-α, TNF-β, TRAILR-1, TRAILR-2, TRAILR-3, TRAILR-4 and XIAP proteins were measured by the membrane array kit. Results: Iris taochia extracts exhibited significant cytotoxic effects on MCF7 cells and IC50 values ranging from 1.56 to 100 μg/mL. Our results indicate that MeOH extract of Iris taochia in MCF7 cells may be a regulator of cell death proteins, cell cycle and growth factors. DCM and EtOH extracts of Iris taochia have a limited effect on MCF7 cells, especially, HSPs, which play a significant role in chemoresistance, downregulating DCM and EtOH extracts of Iris taochia, whereas ligands and receptors of extrinsic apoptotic pathway are upregulated by these extracts. Conclusion: This is the first study to investigate the cytotoxic and apoptotic effect of Iris taochia extracts on MCF7 cells. Results also showed that Iris taochia reduced cell viability and induced apoptotic pathways as a potential regulator of cancer cell death.

In vitro acetylcholinesterase inhibitory activity and chemical composition of the dichloromethane extracts of the leaves and inflorescences of the tumbleweed, Trachyandra laxa (N.E.Br.) Oberm. var. rigida (Suesseng.) Roessl

Trachyandra laxa, which belongs to the family Asphodelaceae, is a poisonous geophyte, often consumed by livestock, particularly when pastures are poor. Ingestion of T. laxa by livestock causes paresis and paralysis of the animals. However, the chemical composition of this plant has never been studied before and its toxic components are still unknown. In this study, an acetylcholinesterase (AChE) inhibition assay of different extracts of the parts of Trachyandra laxa (N.E.Br.) Oberm. var. rigida (Suesseng.) Roessl. revealed that the DCM extracts possess the highest AChE inhibitory activity. In addition, using qualitative phytochemical tests, it was determined that all the plant parts contain polyphenols and tannins, while only the leaves contain flavonoids. Saponins were detected in all plant parts, except in the flowers and anthraquinones were detected in all plant parts expect the roots. Using GC–MS analysis, 13 volatile constituents were identified in the DCM extracts of the leaves and inflorescences, including α,β-unsaturated lactones, carboxylic acids and esters, a diterpenoid, phytol, and a phytosterol, β-sitosterol. Of particular interest are the α,β-unsaturated lactones, which were identified as dihydroactinidiolide (DHA) and its hydroxylated analogue, hydroxy‑DHA, as well as phytol, a diterpenoid. DHA and phytol have been reported to possess potent AChE inhibition activity. It is therefore plausible that these compounds and potentially also hydroxy‑DHA could contribute to the AChE inhibitory activity of the DCM extracts, and thus the neurotoxicity of T. laxa var. rigida.

Cinnamomum zeylanicum extracts reduce lipid oxidation in broadband anchovy (Anchoviella lepidentostole) minced fish

Minced fish (MF) is an interesting material for development of novel fish products; however, the mechanical deboning process interferes with MF quality, by increasing the lipid oxidation. This study reports the effect of Ceylon cinnamon (Cinnamomum zeylanicum) hydro-ethanolic bark extracts as natural antioxidants on both washed and unwashed MF of broadband anchovy during six months of storage at -18 °C. Spray-dried cinnamon extract (DC) and spray-dried cinnamon extract using maltodextrin 10DE (DCM) as a carrier were evaluated in relation to antioxidant activity in vitro. DC and sodium erythobarte were added to both washed and unwashed MF at 0.25% (w:w), whilst DCM was added at 1.055% (w:w). The DC extract presented higher antioxidant activity in vitro compared to DCM extract. The addition of cinnamon extracts reduced the lipid oxidation in washed and unwashed MF compared with the controls. C. zeylanicum extracts can be used to prevent lipid oxidation in MF during the frozen storage and are alternatives for food industries that seek to meet the demand of consumers increasingly concerned with the consumption of healthy foods.

In vitro Cytotoxic Screening of Different Parts from Ornithogalum bungei on Selected Cancer Cells.

Background: Natural products comprise a large section of pharmaceutical agents in the field of cancer therapy. In the present study, the organic extracts and fractions of various parts of Ornithogalum bungei were investigated for in vitro cytotoxic properties on three human cancer cell lines, hepatocellular carcinoma (HepG2), prostate cancer (PC3), and leukemia (K562) cells.Methods: The present experimental study was conducted at Tehran University of Medical Sciences (Tehran, Iran) during 2017-2019. Separately extracted plant materials, including bulbs, stems, and flowers of O. bungei were assessed by the tetrazolium dye-based colorimetric assay (MTT). The selected extracts were submitted to fractionation using vacuum liquid chromatography and after MTT assay, the half maximal inhibitory concentration (IC50 (value for each fraction was determined. The data were analyzed using One-way ANOVA followed by Tukey’s post hoc test. P<0.05 was considered statistically significant.Results: The cytotoxicity of the bulb’s methanol extract and the dichloromethane extract of aerial parts increased in a concentration-dependent manner. Additionally, cell viability decreased in a dose-dependent manner. In the HepG2 cell line, the best IC50 values of fractions from DCM extracts of aerial parts were determined to be 19.8±10.2 µg/mL after 24 hours of exposure and 19.39±6.4 µg/mL following 48 hours of exposure. In the PC3 cell line, after 48 hours of exposure, the IC50 values of fractions were unaccountable, while the percentage of inhibition for A6 to A11 in 24 hours of exposure was more than 40 µg/mL.Conclusion: O. bungei growing in Iran showed significant potentials as a cytotoxic agent with selective effects on different cancer cell lines.

Bio-prospective potential of Pleurotus djamor and Pleurotus florida mycelial extracts towards Gram positive and Gram negative microbial pathogens causing infectious disease

BackgroundThe emergence of resistance to commonly used antibiotics by human infections occurred mostly due to their overuse, that prompted individuals to pursue novel and innovative treatments. The phytochemical characteristics, antibacterial activity, and cytotoxicity of MCF7 cells were evaluated in two Pleurotus spp. mycelial extracts in this work. MethodsPleurotus djamor and Pleurotus florida mycelial extracts from pure cultures were tested for antibacterial activity by a well-diffusion assay and antimicrobial activity against mold fungi was evaluated for biomass inhibition. Mycelial extracts were obtained from dichloromethane extracts and their biophysical characteristics are analyzed by UV–vis spectrum and FTIR analysis. By spraying detection reagents onto TLC plates, the chemicals in dichloromethane extraction of chosen mushroom fungus mycelia were identified. Using the MTT test, the cytotoxic effect of dichloromethane extracts of selected mushroom fungi was evaluated on MCF7 Cell lines. ResultsMycelial extracts of P. djamor and P. florida exhibited significant antimicrobial effect on the bacterial and fungal pathogens tested. Dichloromethane mycelial extracts were obtained using soxhlet extraction which response positive for various phytochemical analysis. Detection of metabolites in thin layer chromatography using spray reagents documented one of few first accounts on flavonoids, anthroquinone and terpenoid compounds in P. djamor and P. florida. P. djamor and P. florida had dose-dependent antiproliferative activity against MCF7 cells, with an inhibitory impact of 55.72% and 64.47% percent at 125 μg/mL, respectively. ConclusionThe study has reported the identification with the potent biological activity of some of the key bioactive components present in DCM extracts from the mycelia of P. djamor and P. florida.

An in vitro study of an Artocarpus heterophyllus substance as a hepatitis C antiviral and its combination with current anti-HCV drugs

BackgroundCurrent therapy of chronic hepatitis C virus (HCV) with direct-acting antivirals (DAAs) has dramatically improved the sustained virologic response (SVR) of affected patients; however, treatment with DAAs remains expensive, and drug-resistant HCV variants remain a threat. As a result, there is still a need to continue to develop affordable and effective drugs for the treatment of HCV. Previously, we have demonstrated that a crude extract from Artocarpus heterophyllus leaves is a potential anti-HCV candidate. In this study, we have further purified this crude extract, examined which sub-fraction possesses the highest antiviral activity, and then explored its efficacy at different HCV life cycle stages. We also assessed synergistic antiviral effects between the A. heterophyllus extract and commercially available anti-HCV drugs.MethodsWe used vacuum liquid chromatography (VLC) and high-performance liquid chromatography (HPLC) to fractionate a dichloromethane extract of A. heterophyllus leaves. We then examined the anti-HCV activity of the fractions using HCV genotype 2a, JFH1a; the antiviral mode of action was determined by exploring adding the treatments at different times. We examined the antiviral effects on the viral entry stage through a virucidal activity test, viral adsorption examination, and pretreatment of cells with the drug. The effects on the post-viral entry stage were determined by the levels of HCV protein expression and HCV RNA expression in infected cells.ResultsThrough activity guided purification, we identified the sub-fraction FR3T3 as possessing the most robust anti-HCV activity with an IC50 value of 4.7 ± 1.0 μg/mL. Mode-of-action analysis revealed that FR3T3 inhibited post-viral entry stages such as HCV NS3 protein expression and HCV RNA replication with marginal effects on the viral entry stage. Thin-layer Chromatography (TLC) indicated that FR3T3 contained terpenoids and chlorophyll-related compounds. We also found a synergistic antiviral activity when the DCM extract of A. heterohyllus was used in combination therapy with commercial anti-HCV drugs; Ribavirin, Simeprevir, Cyclosporin A.ConclusionsThe extract of A. heterophyllus and its sub-fraction, FR3T3, presented here have anti-HCV activities and could be candidate drugs for add-on-therapy for treatment of chronic HCV infections.