Ethyl Acetate Subfraction Research Articles

Assessing the Oxidative Stress Reducing Potential of Spilanthes filicaulis (Schumach & Thonn) Ethyl-Acetate Sub-fractions on Plasmodium berghei Infected Female Mice.

Infections by Plasmodium parasite actuate oxidative stress. As malaria parasite actions overwhelm host antioxidant defense by producing excessive reactive species during haemoglobin degradation. This study aimed to evaluate the oxidative status by considering the antioxidant level of ethyl-acetate sub-fractions of Spilanthes filicaulis (ESSF) extract on Plasmodium berghei NK-65 parasitized mice. Sixty female mice weighing 20.0 ± 3.0g were inoculated intraperitoneally with 0.2mL of parasitized erythrocytes randomly selected into five groups of 12 mice, Groups I and II were orally administered with normal saline (10mL/kg) and chloroquine (10mg/kg) while, Groups III, IV and V were administered 250,500 and 750mg/kg per day respectively of ESSF. Mice in each group were sacrificed on days 4 and 8 post treatment, thereafter blood and liver samples were collected and prepared using standard methods to obtain erythrocytes and liver homogenates. Malondialdehyde (MDA), a measure of lipid peroxidation, superoxide dismutase (SOD) and catalase (CAT) level was assessed in the erythrocyte and liver. Administration of ESSF produced a significant (p < 0.05) decrease in the MDA concentration of the parasitized treated group when compared to parasitized untreated group on day 4. Concomitantly, a significant (p < 0.05) increase in SOD and CAT activity in the treated groups with a corresponding decrease in the untreated group on day 4. However, effects of this action were more pronounced on day 8 in both tissues. These findings suggest that ESSF might contribute to the protection of malaria infected mice against oxidative disruptions by improving antioxidant status.

Cytotoxicity, antiplasmodial and antimalarial effects of the spice and medicinal tree Schinus terebinthifolius

Schinus terebinthifolius is a medicinal plant with biological in vitro activity against clinically relevant parasites such as Leishmania amazonensis and L. chagasi. Considering its antiparasitic activity, this work aimed to investigate the in vitro effects of S. terebinthifolius fractions against P. falciparum using SYBR fluorescence tests, its cytotoxicity towards monkey kidney cells and efficacy against malaria in a murine model of P. berghei infection. For the first time, screening results with several extracts and fractions are reported herein. Five out of 16 fractions obtained from leaves, flowers, unripe and ripe berries with dichloromethane, ethyl acetate and butanol solvents used in plant extraction were active against P. falciparum. Two ethyl acetate fractions with no cytotoxicity in vitro presenting selectivity indexes of ≥111 and ≥167 exhibited no antimalarial activity in mice at 500 mg/kg. Preliminary LCMS analysis of the ethyl acetate subfractions showed antiplasmodial effect in vitro with high specific index suggesting that such activity is related to polyphenolics in the fractions, mainly flavonoids. Additional studies should be undertaken to identify the compound(s) with anti-malaria activity.

Anti-inflammatory and Antinociceptive Activity of Terminalia avicennioides Guill. &amp; Perr. Stem Bark Extract is Distinct from the Antioxidant Effect Mediated by Its Phenolic-rich Fraction

Background: Terminalia avicennioides stem bark has been used in the management of pain and inflammatory disorders in northern Nigeria. This study evaluated the antinociceptive and anti-inflammatory properties of T. avicennioides stem bark extract and its fractions. The correlation of these effects to the presence and concentration of phenolics was also ascertained.&#x0D; Study Design: Experimental Design.&#x0D; Place and Duration of Study: Department of Pharmacology and Toxicology, National Institute for Pharmaceutical Research and Development, Idu, Abuja, Nigeria. The study was conducted from October to December 2022.&#x0D; Methods: A 70%v/v ethanol extract of T. avicennioides stem bark was prepared. Aqueous and ethyl acetate fractions (AF, EF) and sub-fractions (EF1 – EF4) were also prepared. The antioxidant capacity of the extract and fractions were determined using DPPH radical scavenging test, while xylene-induced topical ear edema and formalin test were used to assess antinociceptive and anti-inflammatory activity. The ethyl acetate sub-fractions were further assessed using the egg albumin–induced inflammation. The extract and fractions were characterized by High-Performance Liquid Chromatography (HPLC), and phenolic content was quantified as gallic acid equivalent (GAE)/mg of extract or fraction.&#x0D; Results: The ethyl acetate and aqueous fractions showed higher antioxidant capacity compared to the parent ethanol extract. T. avicennioides ethanol extract significantly inhibited pain (p&lt;0.001) and inflammatory responses (p&lt;0.05) and these effects were more significant compared to those produced by the fractions. Fractions AF and EF exhibited similar activity, although EF produced better inhibition of pain and topical edema. The subfraction EF2 also showed anti-inflammatory activity but this effect was insignificant (p&gt;0.05). The HPLC profiles of the extract and fractions showed peaks corresponding to caffeic acid, chlorogenic acid, and gallic acid. The EF and AF revealed higher peak areas corresponding to gallic acid and rutin respectively. This correlated with a comparatively high gallic acid content and antioxidant effect of the ethyl acetate fraction (GAE: 1.32/mg, IC50 = 0.036 mg/ml), relative to the extract (GAE: 0.88/mg, IC50 = 0.052 mg/ml).&#x0D; Conclusion: The constituents responsible for the antinociceptive and anti-inflammatory activity of T. avicennioides extract appear distinct from antioxidative principles in phenolics-rich fractions.

Anti-diabetic and anti-urease potential of Osbeckia nutans Wall. leaves

A study was conducted to investigate the anti-diabetic and anti-urease potential of Osbeckia nutans leaves (ONL). Six compounds, i.e. quercetin-3-O-glucoside, myricetin, shikimic acid, catechin, trans-ferulic acid and luteolin were identified from the butanol sub-fraction, BE2 and the ethyl acetate sub-fraction, EA5 of ONL. BE2 inhibited α-glucosidase and Jack bean urease with IC50 values of 0.036 μg/mL (437.46 μg/mL for acarbose) and 0.327 mg/mL (0.039 mg/mL for thiourea), respectively. In the glucose uptake experiment, BE2 (0.05 mg/mL) treatment resulted in a substantial increase in glucose uptake in free fatty acid (FFA)-treated cells at a concentration 10 times lower than that seen in EA5 (0.5 mg/mL) treated cells. The binding energies of quercetin-3-O-glucoside with α-glucosidase, glucose transporter GLUT4 and H. pylori urease were found to be −94.2585, −219.8271 and −254.391 kcal/mol, respectively. This study revealed that ONL has anti-diabetic and anti-urease abilities and further in-depth research may unveil its full potential.

Phytochemicals, antimicrobial, and sporicidal activities of moss, Dicranum polysetum Sw., against certain honey bee bacterial pathogens.

Beekeeping is an important agricultural and commercial activity globally practiced. Honey bee is attacked by certain infectious pathogens. Most important brood diseases are bacterial including American Foulbrood (AFB), caused by Paenibacillus larvae (P. larvae), and European Foulbrood (EFB) by Melissococcus plutonius (M. plutonius) in addition of secondary invaders, e.g. Paenibacillus alvei (P. alvei) and Paenibacillus dendritiformis (P. dendritiformis). These bacteria cause the death of larvae in honey bee colonies. In this work, antibacterial activities of extracts, fractions, and isolated certain compounds (nominated 1-3) all originated from moss, Dicranum polysetum Sw. ( D. polysetum), were tested against some honey bee bacterial pathogens. Minimum inhibitory concentration, minimum bactericidal concentration, and sporicidal values ​​of methanol extract, ethyl acetate, and n-hexane fractions ranged between 10.4 and 18.98, 83.4-303.75 & 5.86-18.98µg/mL against P. larvae, respectively. Antimicrobial activities of the ethyl acetate sub-fractions (fraction) and the isolated compounds (1-3) were tested against AFB- and EFB-causing bacteria. Bio-guided chromatographic separation of ethyl acetate fraction, a crude methanolic extract obtained from aerial parts of D. polysetum resulted in three natural compounds: a novel one, i.e. glycer-2-yl hexadeca-4-yne-7Z,10Z,13Z-trienoate (1, dicrapolysetoate; given as trivial name), in addition to two known triterpenoids poriferasterol (2), and γ-taraxasterol (3). Minimum inhibitory concentration ranges were 1.4-60.75, 8.12-65.0, 2.09-33.44 & 1.8-28.75µg/mL for sub-fractions, compounds 1, 2, and 3, respectively.

Solid-state fermentation of Apocynum venetum L. by Aspergillus niger: Effect on phenolic compounds, antioxidant activities and metabolic syndrome-associated enzymes.

This study aimed to evaluate the effect of solid-state fermentation (SSF) with Aspergillus niger on the total phenolic content (TPC), the total flavonoid content (TFC), individual phenolic contents, and antioxidant and inhibitory activities against metabolic syndrome-associated enzymes in an ethanol extract from Apocynum venetum L. (AVL). TPC, TFC, and the contents of quercetin and kaempferol during SSF were 1.52-, 1.33-, 3.64-, and 2.22-fold higher than those of native AVL in the ethyl acetate (EA) subfraction of the ethanol extract. The ABTS·+, DPPH· scavenging, and inhibitory activities against α-glucosidase and pancreatic lipase were found to be highest in the EA subfraction. Fermentation significantly increased the ABTS radical cation, DPPH radical scavenging, and pancreatic lipase inhibitory activities by 1.33, 1.39, and 1.28 times, respectively. TPC showed a significantly positive correlation with antioxidant activities or inhibition against metabolic syndrome-associated enzymes. This study provides a theoretical basis for producing tea products with enhanced antioxidant, antidiabetic, and antihyperlipidemic activities.

Leaves of Chenopodium album as source of natural fungicides against Sclertium rolfsii

Series of experiments were conducted to identify the possible antifungal components of Chenopodium album leaves for the management of a highly destructive soil-borne fungal pathogen Sclerotium rolfsii. A 4% methanolic extract caused up to 82% reduction in biomass of the target organism. This extract was partitioned using solvents of variable polarities, and the obtained subfractions were evaluated for their activity against S. rolfsii. The best antifungal activity was detected for the ethyl acetate subfraction (60–74%) followed by n-hexane (51–69%), n-butanol (50–60%), chloroform (20–40%) while the aqueous sub-fraction had the lowest activity (9–35%) as detected by the decrease in biomass of the pathogen. Ethyl acetate and n-hexane sub-fractions were analyzed for their chemical constituents by GC–MS technique. Literature survey showed that among the identified compounds kitazin P, imidazole-4-carboxylic acid, 2-fluoro-1-methoxymethyl-, ethyl ester and 9,12,15-octadecatrienoic acid, 2,3-dihydroxypropyl ester, (Z,Z,Z)- had antifungal activities against other fungal species and could be responsible for control of S. rolfsii by methanolic extract in the present study.

In vitro antimalarial activity of Syzygium cumini fruit fraction.

Malaria is still one of the most severe public health problems worldwide. The development of treatment, prevention, and control of malaria is one of the substantial problems in the world. To investigate the in vitro antimalarial activity of Syzygium cumini methanol fruit fraction. Syzygium cumini L fruit powder was macerated with methanol (PA) and the extract obtained was fractionated using the liquid-liquid partition method with n-hexane, ethyl acetate, butanol, chloroform, methanol, and water solvents. In vitro antimalarial assay was conducted using the culture of Plasmodium falciparum 3D7 strain culture that had reached >5% growth and was examined for IC50 values using a 24-well microplate in duplicate. Each treatment and control well contained 1,080 μl of complete media. Well, number 1 was added with 120 μl fraction, and then the solution was diluted until it reached 0.01, 0.1, 1, 10, and 100 μg/ml the final concentration in the microtiter well. The control only contained complete media and infected erythrocytes without the addition of anti-malarial drugs. The microplate was incubated for 48 hours. After 48 hours, a thin blood smear was made fixed with methanol and stained with 20% Giemsa for 20 minutes to determine the IC50 value by plotting sample concentrations and percentage of parasitemia in Excel. The IC50 values of ethyl acetate fraction, n.hexane fraction, butanol fraction, and water fraction were 1.189, 76.996, 1,769, and 15.058 μg/ml, respectively. Whereases the IC50 values of C1 fraction (mix fraction from chloroform: methanol 100:0 and 90:10) and C4 fraction (mix fraction from chloroform: methanol 20:80, 10:90, and 0:100) were 100.126 and 1.015 μg/ml, respectively. The results showed that the IC50 value of ethyl acetate, butanol, and C4 fraction were lower than 10μg/ml and were considered as good activity (strong antimalarial activity). The ethyl acetate, butanol and C4 subfraction from S. cumini fruit have the potential to be developed as an antimalarial agent.

Anti-hyperglycemic fraction from Alternanthera sessilis L. leaves gets elucidated following bioassay-guided isolation and mass spectrometry

The anecdotal use of Alternanthera sessilis L. as a relief for diabetes has been known in the Philippines for generations, and antidiabetic activity of similar varieties in other countries is likewise documented. However, the compounds responsible for this activity remain unclear. This study aims to isolate the anti-hyperglycemic fraction of local A. sessilis leaves and identify the compounds in this fraction. Methanol extract of A. sessilis leaves and its hexane, ethyl acetate (ASE), and water fractions were administered to alloxan-induced diabetic mice. ASE (250mg/kg) had the highest anti-hyperglycemic activity at 6-h post-treatment (25.81%±12.72%), with almost similar blood glucose reduction rate as metformin (30.13±3.75%, p=0.767). Repeated fractionation employing chromatographic separation techniques followed by in vivo anti-hyperglycemic assay yielded partially purified subfractions. A. sessilis ethyl acetate subfraction 4-2 (100mg/kg) displayed remarkable suppression of blood glucose rise in diabetic mice at 6-h post-treatment (26.45±3.75%, p<0.0001), with comparable activity with metformin (100mg/kg, 27.87±5.65%, p=0.652). Liquid chromatography/mass spectrometry showed eight distinct peaks, with four peaks annotated via the Traditional Chinese Medicine library and custom library for A. sessilis. Among these, luteolin, apigenin, ononin, and sophorabioside were identified as putative compounds responsible for the anti-hyperglycemic activity. This result provided basis for the reported anecdotal claims and potential utility of the local variety of A. sessilis leaves as sources of anti-hyperglycemic agents.

Abstract : The Effect of 10% ointment concentration of ethyl acetate subfraction of Meniran (Phyllanthus niruri L.) leaves on excision wound healing in white male rats

Abstract : The Effect of 10% ointment concentration of ethyl acetate subfraction of Meniran (Phyllanthus niruri L.) leaves on excision wound healing in white male rats

Bioassay-Guided Isolation of Iridoid Glucosides from Stenaria nigricans, Their Biting Deterrence against Aedes aegypti (Diptera: Culicidae), and Repellency Assessment against Imported Fire Ants (Hymenoptera: Formicidae).

In our natural product screening program, we screened natural products for their repellency and toxicity against insect vectors. Methanolic extract of aerial parts of Stenaria nigricans (Lam.), with no published chemistry, was tested for repellency against mosquitoes and imported hybrid fire ants. Methanolic extracts showed biting deterrence similar to DEET (N,N-diethyl-3-methylbenzamide) against Aedes aegypti L. Based on this activity, the crude extract was fractionated into chloroform, ethyl acetate, and methanol subfractions. The active methanolic subfraction was further fractionated into 13 subfractions. These fractions were tested for their biting deterrence against Ae. Aegypti. Active subfractions were further characterized to identify the compounds responsible for this activity. Four undescribed iridoid glucosides (1–4) and three previously reported compounds (5–7) were isolated from active subfractions and tested for their biting deterrent activity. Based on BDI values, compounds 2, 3, 6, and 7, with biting deterrence similar to DEET, showed the potential to be used as repellents against mosquitoes. In an in vitro digging bioassay, none of these compounds showed any repellency against hybrid imported fire ants at a dose of 125 µg/g. This is the first report of biting deterrence and repellency of S. nigricans extract and its pure compounds, iridoid glucosides against mosquitoes and imported fire ants. Further studies will be conducted to explore the repellent potential of these compounds in different formulations under field conditions.

Phytochemical Composition and Biological Activities of Arctium minus (Hill) Bernh.: A Potential Candidate as Antioxidant, Enzyme Inhibitor, and Cytotoxic Agent

Arctium minus (Hill) Bernh. (Asteraceae), which has a wide distribution area in Turkey, is a medicinally important plant. Eighty percent methanol extracts of the leaf, flower head, and root parts of A. minus were prepared and their sub-fractions were obtained. Spectrophotometric and chromatographic (high-performance liquid chromatography) techniques were used to assess the phytochemical composition. The extracts were evaluated for antioxidant activity by diphenyl-2-picrylhydrazil radical (DPPH●), 2,2′-Azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS●+) radical scavenging, and β-carotene linoleic acid bleaching assays. Furthermore, the extracts were subjected to α-amylase, α-glucosidase, lipoxygenase, and tyrosinase enzyme inhibition tests. The cytotoxic effects of extracts were investigated on MCF-7 and MDA-MB-231 breast cancer cell lines. The richest extract in terms of phenolic compounds was identified as the ethyl acetate sub-fraction of the root extract (364.37 ± 7.18 mgGAE/gextact). Furthermore, chlorogenic acid (8.855 ± 0.175%) and rutin (8.359 ± 0.125%) were identified as the primary components in the leaves’ ethyl acetate sub-fraction. According to all methods, it was observed that the extracts with the highest antioxidant activity were the flower and leaf ethyl acetate fractions. Additionally, ABTS radical scavenging activity of roots’ ethyl acetate sub-fraction (2.51 ± 0.09 mmol/L Trolox) was observed to be as effective as that of flower and leaf ethyl acetate fractions at 0.5 mg/mL. In the β-carotene linoleic acid bleaching assay, leaves’ methanol extract showed the highest antioxidant capacity (1422.47 ± 76.85) at 30 min. The enzyme activity data showed that α-glucosidase enzyme inhibition of leaf dichloromethane extract was moderately high, with an 87.12 ± 8.06% inhibition value. Lipoxygenase enzyme inhibition was weakly detected in all sub-fractions. Leaf methanol extract, leaf butanol, and root ethyl acetate sub-fractions showed 99% tyrosinase enzyme inhibition. Finally, it was discovered that dichloromethane extracts of leaves, roots, and flowers had high cytotoxic effects on the MDA-MB-231 cell line, with IC50 values of 21.39 ± 2.43, 13.41 ± 2.37, and 10.80 ± 1.26 µg/mL, respectively. The evaluation of the plant extracts in terms of several bioactivity tests revealed extremely positive outcomes. The data of this study, in which all parts of the plant were investigated in detail for the first time, offer promising results for future research.

Antiviral activity of Cenostigma pluviosum var. peltophoroides extract and fractions against SARS-CoV-2

Few extracts of plant species from the Brazilian flora have been validated from a pharmacological and clinical point of view, and it is important to determine whether their traditional use is proven by pharmacological effects. Cenostigma pluviosum var. peltophoroides is one of those plants, which belongs to the Fabaceae family that is widely used in traditional medicine and is very rich in tannins. Due to the lack of effective drugs to treat severe cases of Covid-19, the main protease of SARS-CoV-2 (Mpro) becomes an attractive target in the research for new antivirals since this enzyme is crucial for virus replication and does not have homologs in humans. This study aimed to prospect inhibitor candidates among the compounds from C. pluviosum extract, by virtual screening simulations using SARS-CoV-2 Mpro as target. Experimental validation was made by inhibitory proteolytic assays of recombinant Mpro and by antiviral activity with infected Vero cells. Docking simulations identify four compounds with potential inhibitory activity of Mpro present in the extract. The compound pentagalloylglucose showed the best result in proteolytic kinetics experiments, with suppression of recombinant Mpro activity by approximately 60%. However, in experiments with infected cells ethyl acetate fraction and sub-fractions, F2 and F4 of C. pluviosum extract performed better than pentagalloylglucose, reaching close to 100% of antiviral activity. The prominent activity of the extract fractions in infected cells may be a result of a synergistic effect from the different hydrolyzable tannins present, performing simultaneous action on Mpro and other targets from SARS-CoV-2 and host. Communicated by Ramaswamy H. Sarma

In Vitro Antioxidant and Anticancer Properties of Various E. senegalensis Extracts.

Although Erythrina senegalensis is a plant widely used in traditional medicine in sub-Saharan Africa, its biological properties have been poorly investigated to date. We first characterized by conventional reactions the composition of several stem bark extracts and evaluated in acellular and cellular assays their pro- or antioxidant properties supported by their high phenolic and flavonoid content, particularly with the methanolic extract. The pro- or antioxidant effects observed did not correlate with their IC50 concentrations against five cancer cell lines determined by MTT assay. Indeed, the CH2Cl2 extract and its ethyl acetate (EtOAc) subfraction appeared more potent although they harbored lower pro- or antioxidant effects. Nevertheless, at equipotent concentration, both extracts induced ER- and mitochondria-derived vacuoles observed by fluorescent microscopy that further led to non-apoptotic cell death. LC coupled to high resolution MS investigations have been performed to identify chemical compounds of the extracts. These investigations highlighted the presence of compounds formerly isolated from E. senegalensis including senegalensein that could be retrieved only in the EtOAc subfraction but also thirteen other compounds, such as 16:3-Glc-stigmasterol and hexadecanoic acid, whose anticancer properties have been previously reported. Nineteen other compounds remain to be identified. In conclusion, E. senegalensis appeared rich in compounds with antioxidant and anticancer properties, supporting its use in traditional practice and its status as a species of interest for further investigations in anticancer drug research.

LC-MS/MS metabolomics-facilitated identification of the active compounds responsible for anti-allergic activity of the ethanol extract of Xenostegia tridentata.

In vivo and in vitro anti-allergic activities of ethanol extract of Xenostegia tridentata (L.) D.F. Austin & Staples were investigated using passive cutaneous anaphylaxis reaction assay and RBL-2H3 cell degranulation assay, respectively. The crude ethanol extract exhibited promising activities when compared with the known anti-allergic agents, namely dexamethasone and ketotifen fumarate. The ethyl acetate subfraction showed the highest anti-allergic activity among various sub-partitions and showed better activity than the crude extract, consistent with the high abundance of total phenolic and flavonoid contents in this subfraction. LC-MS/MS metabolomics analysis and bioassay-guided isolation were then used to identify chemical constituents responsible for the anti-allergic activity. The results showed that major components of the ethyl acetate subfraction consist of 3,5-dicaffeoylquinic acid, quercetin-3-O-rhamnoside, kaempferol-3-O-rhamnoside and luteolin-7-O-glucoside. The inhibitory activity of the isolated compounds against mast cell degranulation was validated, ensuring their important roles in the anti-allergic activity of the plant. Notably, besides showing the anti-allergic activity of X. tridentata, this work highlights the role of metabolomic analysis in identifying and selectively isolating active metabolites from plants.

Anti-Hyperuricemic Effect of Ethyl Acetate Sub-Fractions from Chrysanthemum morifolium Ramat. Dried Flowers on Potassium Oxonate-Induced Hyperuricemic Rats

Xanthine oxidase (XO) plays an important role in purine degradation in humans. The study aimed to determine the XO inhibitory potential of Chrysanthemum morifolium dried flower ethyl acetate sub-fractions and its anti-hyperuricemic effect in rat models. Bioassay-guided fractionation based on XO inhibitory assay was employed to obtain bioactive fractions and sub-fractions. In vitro cytotoxicity and cellular antioxidant capacity of the sub-fraction and its mode of XO inhibition were also investigated. The anti-hyperuricemic effect of the bioactive sub-fraction was investigated using rat models via oral consumption, and followed by an XO mRNA gene expression study. The compounds in the bioactive sub-fractions were identified putatively using HPLC-Q-TOF-MS/MS. Ethyl acetate (EtOAc) fraction exhibited the highest XO inhibition among the fractions. It was further fractionated into 15 sub-fractions. F10 exhibited high XO inhibitory activity, cellular pro-proliferative effect, and intracellular antioxidant activity among the sub-fractions tested. This sub-fraction was non-cytotoxic at 0.1–10 µg/mL, and very effective in lowering serum and urine uric acid level in rat models upon oral consumption. A total of 26 known compounds were identified and seven unknown compounds were detected via HPLC-Q-TOF–MS/MS analysis. The possible mechanisms contributing to the anti-hyperuricemic effect were suggested to be the non-competitive inhibition of XO enzyme, XO gene expression down-regulation, and the enhancement of uric acid excretion.

Bioassay Guided Fractionation of Phytocompounds from Bergenia ligulata: A synergistic approach to treat drug resistant bacterial and fungal pathogens

Bergenia ligulata Wall. Engl. is perennial herb used for the treatment urinary discharges, diseases of urinary bladder, dysentery, menorrhagia, splenic enlargement and heart diseases. The aim of the present study was to investigate the antimicrobial and synergistic potential of solvent fractions and major phytocompound of B. ligulata against bacterial and fungal pathogens. The antimicrobial and synergistic potential of the solvent fractions (Petroleum ether, chloroform, ethyl acetate, and remaining aqueous) of methanolic extract of B. ligulata rhizome and major phytocompound bergenin were investigated by agar well diffusion and broth microdilution method. Major Phytocompound bergenin was quantified in solvent fractions of B. ligulata by HPTLC and HP-LC method Molecular docking of bergenin was done by AutoDock vina with bacterial and fungal targets. Drug likeness and ADME screening was done with help of molinspiration and swissADME servers. Ethyl acetate sub-fraction of B. ligulata showed the highest antimicrobial activity against bacterial and fungal strains with zone of inhibition 13, 13, 14, 13 mm against Escherichia coli, Staphylococcus aureus, Candida albicans MTCC 277 and ATCC 90028 respectively. The MIC of ethyl acetate sub-fraction against E. coli, S. aureus, C. albicans MTCC 277 and ATCC 90028 was 31.25, 250, 62.5, 62.5 μg mg−1 respectively. Bergenin major phytocompound of B. ligulata also showed the antimicrobial activity with MIC 250 μg mg−1 against all the selected bacterial and fungal strains. Quantification of bergenin showed that ethyl-acetate sub fraction (17.6 μg mg−1 and 112 μg mg−1 measured through HPTLC and HPLC, respectively) is rich in bergenin content. The bergenin content recovered in ethyl acetate sub-fraction is 38.2%. Bergenin showed good interactions with bacterial and fungal targets and follows all parameters of drug likeness. Conclusions: Solvent fractions of B. ligulata and major phytocompound Bergenin can be used as a bioenhancer of antibacterial and antifungal agents to treat drug resistant pathogens.

Evaluation of cytotoxic activity of Fomes fomentarius extracts against brine shrimp larvae and colon adenocarcinoma cell line (SW742)

Aim: Cancer is one of the leading worldwide public health problems. Natural product has provided a valuable source biological active compound for maintaining humans health even after years of introduction of synthetic drugs. Fomes fomentarius (F. fomentarius), a fungus belongs to Polyporaceae family, is widely used for different medicinal purposes. Considering inadequate reports on the fungus, cytotoxic effect of its extracts on brine shrimp larvae and colon adenocarcinoma cell line (SW742) was investigated in this study. Methods: The fruiting body of the fungus was chopped and extracted with water, ethanol, methanol, and ethanol-methanol (50-50) solvents. Regarding the results of brine shrimp lethality assay (BSLA), ethanol extract showed the highest toxicity, so subjected to fractionation with chloroform, ethyl acetate, and water. The fractions were tested for cytotoxic activity in human colon adenocarcinoma (SW742) cells by MTT assay. Ethyl acetate fraction exhibited more cytotoxic activity against SW742 cell line. Therefore, it was chosen to be purified using silica gel column chromatography. Subfractions of ethyl acetate were tested for their cytotoxicity by MTT assay. The content of these subfractions were also analyzed using gas chromatography-mass spectrometry (GC-MS). Results: According to the results, ethanol extract had the greatest potency in BSLA, and ethyl acetate fraction showed the highest cell cytotoxicity. Based on GC-MS chromatogram, palmitic acid was the compound with the largest quantity in the three selected subfractions of ethyl acetate. Conclusion: It was found that palmitic acid with other compounds in F. fomentarius had cytotoxic activity against colon adenocarcinoma cells. This feature may provide a biological basis for further clinical evaluation.

Determination of the rate of kill, mode of action, and the bioactive components from the ethyl acetate sub-fraction of methanol extract of Phyllanthus amarus.

The time-kill rate of methanol extract of Phyllanthus amarus was determined in this study which showed that the extract caused a reduction of the viable cells of all the test bacteria after a contact time of 30 mins and there were virtually no surviving cells of all the test bacteria after a contact time of 180 mins. The extract was also found to cause leakages of cellular materials such as potassium ions, sodium ions, protein and nucleic acids from the test bacteria which led to the loss of cell viability. The ethyl acetate sub-fraction of the extract was analyzed by GC-MS and FTIR analysis and the result revealed the presence of Phytochemicals such as 1, 2-Benzenedicarboxilic acid mono (2-Ethylhexyl) ester, Columbin, 2-(6-Methylpyridin-2-ylmethyl) cyclohexane, 2(1H) Naphthalenone,3,5,6,7,8,8a-hexahydro-4,8a-dimethyl-6-(1-methylethenyl) which have all been reported to possess antibacterial activity against both gram-positive and gram-negative bacteria. The result of this study will contribute to the baseline data on the pharmacodynamics of the extract if applied as herbal medicine for human treatment thereby reducing the dosage and period of treatment. The finding also revealed that the ethyl acetate sub-fraction of methanol extract of P. amarus contains antibacterial phytochemicals that may be used to develop more potent, safe and cheap antimicrobial agents using nanotechnology.&#x0D;

Chemical Components of Polygonum cuspidatum Ethylacetate Subfraction and their Effects on Epstein-Barr Virus Lytic Genes Expression

The study highlights the effect of ethylacetate subfraction F1 from Polygonum cuspidatum root and piceid on the inhibition of Epstein-Barr virus (EBV) lytic gene expression. The MTT [3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyltetrazolium bromide] method was employed to assess the effect of F1 and piceid on the cell survival of P3HR1. The immunoblotting method and flow cytometry were applied to analyze the expression of EBV lytic proteins. The results showed that the EC50 (effective concentration re