Ethyl Acetate Fraction Of Extract Research Articles

Khasiat dan Profil Kromatogram Fraksi Aktif dari Ekstrak Kulit Buah Manggis (Garcinia mangostana L.) yang Diiradiasi

The mangosteen fruit peel (Garcinia mangostana L.) are used as anti-infl ammatory, antihistamine, treatment of heart disease, antibacterial, antifungal, it is also used for the treatment or therapy of cancer, because it has a cytotoxic activity against cancer cells. Most of Indonesian people have used the mangosteen fruit peel and they produce mangosteen peel into powder or herbal medicine industry has produced its extract in capsules. Efforts to preserve dried mangosteen rind has done by heating process or by gamma irradiation technique. This study aimed to study the effect of gamma irradiation on cytotoxicity activity of extracts and active fractions mangosteen fruit peel against L1210 leukemia cells and chromatogram profi le of extracts and active fractions of the mangosteen fruit peel. Dry powder mangosteen rind irradiated using gamma irradiation dose of 5; 7.5; 10; and 15 kGy. Then each sample was successive macerated in n-hexane, ethyl acetate, and ethanol. Each extract was tested cytotoxicity against L1210 leukemia cells. The ethyl acetate extract was most active extract (IC50 = 4.17 mg/mL) compared with n-hexane extract (IC50 = 8.29 mg/mL) and ethanol extract (IC50 = 7.52 mg/mL). Fractionation of ethyl acetate extract by column chromatography were obtained 6 fractions. The result of cytotoxicity test showed that fraction 1 was the most active fraction (IC50 = 3.97 mg/mL), it was it was still categorized as potential anticancer (IC50 ≤ 20 mg/mL). Profi le chromatogram of fraction 1 with TLC-densitometry showed patches of discoloration on irradiation dose of 10 and 15 kGy. The results of analysis by HPLC fraction 1 showed a decrease of peak area at a dose of 10 kGy was signifi cantly different from the control. Based on the chromatogram profi le of fraction 1 and itscytotoxicity against L1210 leukemia cells, so the maximum dose of 7.5 kGy gamma irradiation can beap plied on irradiation of mangosteen fruit peel without changing its effi cacy as anti-cancer agent.

Hepatoprotective Activity of Ethyl Acetate Fraction of Senggugu’s Root Bark (Clerodendrum serratum L.Moon) on Rats Induced by CCl4

Senggugu is a plant that has long been used to treat syphilis, typhoid, cancer, jaundice, and hypertension. The pharmacological activity of senggugu in Indonesia that have been reported include antifertility activity in leaves, mucolytic activity, anti-inflammatory and tracheospasmolytic, also antioxidant in its root bark. This study aims to determine the hepatoprotective effect of ethyl acetate extract fraction of senggugu’s root bark in rats induced by CCl 4 . The powder of senggugu’s root bark was extracted by terraced maceration method starts from n -hexane, ethyl acetate, to methanol, thus obtained ethyl acetate extract fraction of senggugu’s root bark (FEAKAS). The ethyl acetate extract fractions were then tested for hepatoproctective activity using doses of 25, 50, and 100 mg/Kg.BW on rats induced by CCl 4 . FEAKAS hepatoprotective activity was determined from the analysis of blood biochemical and oxidative stress parameters. The blood biochemical parameters included SGOT (serum glutamic oxaloacetic transaminase), SGPT (serum glutamic pyruvic transaminase), ALP (alkaline phosphatase), bilirubin, and total protein were measured with test kit. The oxidative stress parameters were measured from homogenates of liver tissue that were prepared by adding 500 mL of 50 nM Tris buffer (pH 7.4) containing 1 mM EDTA and 10 µg/mL leupeptin. The homogenates were centrifuged to obtain supernatants for measurement of oxidative stress parameters using spectrophotometer method, including MDA (malondialdehyde), GPx (glutathione peroxidase) and CAT (catalase). The results showed that the effect of FEAKAS against CCl 4 induction for preventing lipid peroxidation, from both blood chemical and oxidative stress parameters, are shown at a dose of 100 mg/Kg.BW that significantly different compared to CCl 4 control (ρ <0.05) on all blood chemical and oxidative stress parameters.

Apigenin-7-O-Β-D-Glucuronide Methyl Ester Isolated from Manilkara zapota Leaves

In nature there are numerous plants available with medicinal properties. Around 70 % of medicinal plants are found in tropical areas of India. The search for natural products and compounds derived from natural sources has played a vital role in drug discovery due to their pharmacological importance. Compounds isolated from botanical sources remain an important source of several clinically useful anti-inflammatory agents. Manilkara zapota is a large, evergreen forest tree belongs to family sapotaceae. It is commonly known as chiku (Hindi), sofeda (Bengali) sapodilla and sapoti (French), chickle tree, hase berry, tree potato (English). Manilkara zapota is a species of lowland rain forest. Manilkara zapota and its different parts have been traditionally used for alleviating inflammation related diseases such as arthritis, cancer and skin infections. The present study aims to isolate, structurally characterize and analyze the bioactive compound from Manilkara zapota by using chromatographic and spectrophotometric techniques on the basis of inhibitory effects on sPLA2 by activity guided fractionation of ethyl acetate extract of Manilkara zapota leaves. Among the six fractions (F1-F6) tested fraction-5 showed significant inhibitory effects on sPLA2 activity hence fraction -5 was further subjected to structural analysis for identification of bioactive compounds by using analytical techniques such as TLC, HPLC, FT-IR, LC-MS and 1H ,13C NMR studies. The isolated compound identified as apigenin-7-O-β-D-glucuronide methyl ester.

Penelusuran aktivitas antibakteri, antioksidan, dan sitotoksik fungi endofit genus fusarium sp. diisolasi dari daun meniran (Phyllantus niruri Linn.)

Endophytic fungi used as a new resource to produce bioactive compounds such as antibacterial, antioxidant and anticancer. This study aimed to analyze the antibacterial, antioxidant, cytotoxic fraction of the separation ethyl acetate extract of mycelium of endophytic fungi genus Fusarium sp. Diethyl ether fraction showed high antibacterial activity in E.coli (20,75 µg/mL), S.typhi (35,08 µg/mL), and S.aureus (51,96 µg/mL). Test showed that the antioxidant activity of ethanol 96% fraction has highest antioxidant activity and total phenolic content (75,85 ± 0.87 mg GAE). Cytotoxic test on T47D breast cancer cells showed that the fraction of diethyl ether have highest activity with IC 50 of 10,16 ± 0,88 µg /mL.

In vitro antioxidant assessment and a rapid HPTLC bioautographic method for the detection of anticholinesterase inhibitory activity of Geophila repens

ObjectiveGeophila repens (L.) I. M. Johnst. (Rubiaceae), a small, creeping, perennial herb, is claimed to have memory-enhancing property. The goal of this study was to assess its antioxidant and anticholinesterase activity and conduct a rapid bioautographic enzyme assay for screening acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition of G. repens extracts. MethodsAntioxidant activity of G. repens extracts was assessed by performing 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide (NO), superoxide (SOD), hydroxyl (OH) and total antioxidant capacity (TAC) assays. Anticholinesterase activity was investigated by quantifying the AChE and BChE inhibitory activities of chloroform (CGR), ethyl acetate (EGR) and methanol (MGR) extract fractions from G. repens leaves. A rapid high-performance thin-layer chromatography (HPTLC) bioautographic method for the detection of AChE and BChE inhibition was performed. ResultsAmong all extract fractions, EGR exhibited the highest half maximal inhibitory concentration (IC50) in DPPH, SOD, NO, OH and TAC assays, with IC50 of (38.33 ± 3.21), (45.14 ± 1.78), (59.81 ± 1.32), (39.45 ± 0.79) and (43.76 ± 0.81) µg/mL respectively. EGR displayed competitive, reversible inhibition of AChE and BChE activities with IC50 of (68.63 ± 0.45) and (59.45 ± 0.45) µg/mL, respectively. Total phenolic and flavonoids contents of EGR were found to be 360.42 mg gallic acid equivalents and 257.31 mg quercetin equivalents per gram of extract. Phytoconstituents of the EGR extract that were inhibitors of cholinesterase produced white spots on the yellow background of HPTLC plates in the bioautographic test. ConclusionThe results of this study revealed that phenols and flavonoids could be responsible for the antioxidant, anticholinesterase activities of G. repens.

Advancing HPLC-PDA-HRMS-SPE-NMR Analysis of Coumarins in Coleonema album by Use of Orthogonal Reversed-Phase C18 and Pentafluorophenyl Separations.

A hyphenated procedure involving high-performance liquid chromatography, photodiode array detection, high-resolution mass spectrometry, solid-phase extraction, and nuclear magnetic resonance spectroscopy, i.e., HPLC-PDA-HRMS-SPE-NMR, has proven an effective technique for the identification of compounds in complex matrices. Most HPLC-PDA-HRMS-SPE-NMR investigations reported so far have relied on analytical-scale reversed-phase C18 columns for separation. Herein is reported the use of an analytical-scale pentafluorophenyl column as an orthogonal separation method following fractionation of a crude ethyl acetate extract of leaves of Coleonema album on a preparative-scale C18 column. This setup allowed the HPLC-PDA-HRMS-SPE-NMR analysis of 23 coumarins, including six new compounds, 8-O-β-d-glucopyranosyloxy-6-(2,3-dihydroxy-3-methylbut-1-yl)-7-methoxycoumarin (4), (Z)-6-(4-β-d-glucopyranosyloxy-3-methylbut-2-en-1-yl)-7-hydroxycoumarin (6), 6-(4-β-d-glucopyranosyloxy-3-methylbut-1-yl)-7-hydroxycoumarin (8), (Z)-7-(4-β-d-glucopyranosyloxy-3-methylbut-2-en-1-yloxy)coumarin (13), (S)-8-(3-chloro-2-hydroxy-3-methylbut-1-yloxy)-7-methoxycoumarin (19), and 7-(3-chloro-2-hydroxy-3-methylbut-1-yloxy)coumarin (20). The use of the pentafluorophenyl column even allowed separation of several regioisomers that are usually difficult to separate using reversed-phase C18 columns. The phytochemical investigation described for C.album in this report demonstrates the potential and wide applicability of HPLC-PDA-HRMS-SPE-NMR for accelerated structural identification of natural products in complex mixtures.

Ultrasound-assisted enzyme catalyzed hydrolysis of olive waste and recovery of antioxidant phenolic compounds

Abstract The ultrasound-assisted enzyme hydrolysis (UAEH) procedure for extraction of phenolics from olive waste (OW) was established. The optimized conditions obtained with response surface methodology (RSM) were pH 5.75, treatment 40 min and temperature 55 °C, the yield obtained reached to 4.0%. The extraction kinetics model was fitted well to the ln ( C t / C 0 ) = kt , and the rate constants ( k ) increased with the increased in the extraction temperature. Moreover, the values of k were higher and the activation energy was lower than ultrasound treatment alone. The thermodynamic parameters indicated the extraction process was easier under the UAEH. Compared with the petroleum ether and n-butyl alcohol, ethyl acetate extract fraction had high phenolic content (64.4 mg/g), especially hydroxytyrosol and tyrosol (17.2 mg/g and 14.1 mg/g), and exhibited excellent antioxidant activities in DPPH·, ABTS + ·and FRAP assay. These finding revealed that OW could be used for a enrich source of natural phenolic compounds for development of food industry. Industrial relevance Olive waste (OW) has been widely paid attention as sources of high-added value compounds, such as phenolic which can be obtained by the several methods. And phenolic extract as a food additive can not only improve the sensory quality of food, but also can enhance antioxidant capacity in fatty food matrices. Moreover, the use of this extract has better economic benefits than synthetic additives usually used in food industry. In this study, a novel green, simple and efficient technology was employed to extract the phenolic compounds, which was suitable for large-scale industrial use. The results indicated that ultrasound-assisted enzyme hydrolysis (UAEH) has significant potential in the extraction process of phenolics from OW exhibiting higher phenolic yield in shorter extraction times.

English

Azadirachta indica A. Juss, known as neem (Meliacea family), has insecticide and pesticide properties, and many studies have shown their efficacy as antifungal, anti-inflammatory, among others. Studies for the development of drugs from plants are rising due to several factors such as bacterial resistance, indiscriminate use and the adverse reactions of antibiotics. In this study, phytochemical triage and thin layer chromatography analysis were performed, with similar results as the presence of flavonoids, tannins and terpenes. The antimicrobial activity showed that the ethyl acetate extract and butanol fraction presented greater activity against Streptococcus mutans and Streptococcus mitis presenting a MIC = 50 µg/ml for these strains, and the strain Enteroccocus faecalis, the hydroethanolic extract and aqueous fraction were most promising samples with a MIC = 50 µg/ml and MIC = 25 µg/ml, respectively. Therefore, it encourages the continuation of studies, aiming at the development of cosmetics or toothpaste. Key words: Antimicrobial activity, Azadirachta indica, minimum inhibitory concentration, oral strains, phytochemistry.

Bioassay-guided fractionation of ethyl acetate extract from Armillaria mellea attenuates inflammatory response in lipopolysaccharide (LPS) stimulated BV-2 microglia

BackgroundArmillaria mellea (A. mellea) is a traditional Chinese medicinal and edible mushroom, which is proved to possess a lot of biological activities, including anti-oxidation, immunopotentiation, anti-vertigo and anti-aging activities. However, little information is available in regard to its neuroprotection activity in inflammation-mediated neurodegenerative diseases. PurposeWe have found that A. mellea has an anti-inflammatory activity in LPS-induced RAW264.7 cells in our previous study. The objective of this study is to investigate the anti-neuroinflammatory mechanism of a bioassay-guided fractionation (Fr.2) and its active components/compounds. MethodsCompounds were isolated by preparative high performance liquid chromatography (pre-HPLC) and their structures were established by mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopic analyses. The anti-neuroinflammatory effect of Fr.2 and each compounds were investigated in lipopolysaccharide (LPS)-stimulated murine microglia cell lineBV-2. ResultsWe demonstrated that Fr.2 significantly decreased the production of inflammation mediator nitric oxide (NO) and inflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1beta (IL-1β) in a dose-dependent manner (10, 30, 100µg/ml). In addition, Fr.2 markedly down-regulated the phosphorylation levels of nuclear factor kappa B p65 (NF-κB p65), inhibitory κB-α (IκB-α) and c-Jun N-terminal kinases (JNKs) pathways. Sevens compounds were isolated from Fr.2, among them, three compounds, 5-hydroxymethylfurfural (CP1), vanillic acid (CP4) and syringate (CP5) were reported for the first time in A. mellea. NO and inflammatory cytokines (TNF-α, IL-6, IL-1β) secretion indicated that daidzein (CP6) and genistein (CP7) showed a more outstanding anti-inflammation potential at non-toxic concentrations (10, 30, 100µM) than the other five compounds. ConclusionsIn conclusion, Fr.2 may have therapeutic potential for neurodegenerative diseases by inhibiting inflammatory mediators and suppress inflammation pathway in activated microglia. Daidzein and genistein may serve as the effective anti-inflammation compounds of Fr.2.

Photoprotective effects of cranberry juice and its various fractions against blue light-induced impairment in human retinal pigment epithelial cells

Context: Cranberry has numerous biological activities, including antioxidation, anticancer, cardioprotection, as well as treatment of urinary tract infection (UTI), attributed to abundant phenolic contents.Objective: The current study focused on the effect of cranberry juice (CJ) on blue light exposed human retinal pigment epithelial (ARPE-19) cells which mimic age-related macular degeneration (AMD).Materials and methods: Preliminary phytochemical and HPLC analysis, as well as total antioxidant capacity and scavenging activity of cranberry ethyl acetate extract and different CJ fractions (condensed tannins containing fraction), were evaluated. In cell line model, ARPE-19 were irradiated with blue light at 450 nm wavelength for 10 h (mimic AMD) and treated with different fractions of CJ extract at different doses (5–50 μg/mL) by assessing the cell viability or proliferation rate using MTT assay (repairing efficacy).Results: Phytochemical and HPLC analysis reveals the presence of several phenolic compounds (flavonoids, proanthocyanidin, quercetin) in ethyl acetate extract and different fractions of CJ. However, the condensed tannin containing fraction of ethyl acetate extract of CJ displayed the greater (p < 0.05) scavenging activity especially at the dose of 1 mg/mL. Similarly, the condensed tannin containing fraction at 50 μg/mL presented better (p < 0.05) repairing ability (increased cell viability). Furthermore, the oligomeric condensed tannin containing fraction display the best (p < 0.05) repairing efficiency at 50 μg/mL.Discussion and conclusion: In conclusion, this study distinctly proved that condensed tannin containing fraction of CJ probably exhibits better free radicals scavenging activity and thereby effectively protected the ARPE-19 cells and thus, hampers the progress of AMD.

Antimicrobial Activity of Fractions of Ethyl Acetate Extract of Cladosporium oxysporum, An Endophytic Fungus Derived from Alyxia reinwardtii

The endophytic fungi Cladosporium oxysporum was isolated from medicinal plants Alyxia reinwardtii. The antimicrobial activity of fractions of ethyl acetate extract and also antioxidant and antimicrobial activities of fraction 7 of ethyl acetate extract of (AR-7) were determined in this study. Antimicrobial activity was determined by TLC bioautography and disc diffusion methods. Antioxidant activity was determined by TLC-DPPH autography assay. The fractions of ethyl acetate extract of C. oxysporum at dose of 100 mg/ml were active as antimicrobial against S. aureus, E. coli and C. albicans in disc diffusion method test. Fraction AR-7 was active as antioxidant in TLC-DPPH autography assay. Fraction AR-7 was active as antimicrobial against S. aureus, but was not active against another tested microoorganisms in TLC-bioautography

6′-O-Caffeoyldihydrosyringin isolated from Aster glehni suppresses lipopolysaccharide-induced iNOS, COX-2, TNF-α, IL-1β and IL-6 expression via NF-κB and AP-1 inactivation in RAW 264.7 macrophages

Previously, we found that ethyl acetate extract fraction of Aster glehni exhibited anti-hyperuricemic effects in animal models and also five new caffeoylglucoside derivatives were isolated from this fraction. In this work, we evaluated the anti-inflammatory effects of these caffeoylglucoside derivatives and found that 6′-O-caffeoyldihydrosyringin (2, CDS) most potently inhibited the LPS-induced production of nitric oxide (NO) and prostaglandin E2 (PGE2) in RAW 264.7 macrophages. In addition, CDS was found to concentration-dependently reduce the production of NO, PGE2, and the pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β) induced by LPS in macrophages. Consistent with these observations, CDS concentration-dependently inhibited LPS-induced inducible nitric oxide synthase (iNOS) and cyclooxidase-2 (COX-2) expression at the protein level and also iNOS, COX-2, TNF-α, and IL-6, IL-1β expression at the mRNA level. Furthermore, CDS suppressed the LPS-induced transcriptional activities of nuclear factor-κB (NF-κB) and activator protein-1 (AP-1) as well as the phosphorylation of p65 and c-Fos. Taken together, these results suggest that the anti-inflammatory effect of CDS is associated with the downregulation of iNOS, COX-2, TNF-α, IL-1β, and IL-6 expression via the negative regulation of NF-κB and AP-1 activation in LPS-induced RAW 264.7 macrophages.

Anti-oxidative and anti-neuroinflammatory effects of ethyl acetate extract fractions of &lt;i&gt;Suaeda asparagoides&lt;/i&gt; MIq

Purpose : To evaluate the in vitro antioxidant and anti-neuroinflammatory effects of Suaeda asparagoides ethylacetate extract (SAE) in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Methods : The antioxidative activity of SAE was evaluated by measuring 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity spectrometrometrically. Cell viability was evaluated by 3-(4, 5- dimethylthiazol-2-yl)-2, 5- diphenyl-tetrazolium bromide (MTT) assay, while LPS-stimulated BV-2 microglia were used to study the expression and production of inflammatory mediators, including nitric oxide (NO), inducible NO synthase (iNOS) and tumor necrosis alpha (TNF-α). Results : Pretreatment with SAE prior to LPS treatment significantly inhibited excessive production of NO (p < 0.001 at 20, 40, 80 and 100 μg/mL) in a dose-dependent manner, and was associated with down-regulation of expression of inducible nitric oxide synthase (iNOS). SAE also suppressed the LPSinduced increase in TNF-α level (p < 0.01at concentrations of 40 and 80 μg/mL) in BV-2 cells. Furthermore, DPPH-generated free radicals were inhibited by SAE in a concentration-dependent manner. Conclusion : These results indicate that SAE possesses strong anti-oxidant properties, and inhibits excessive production of pro-inflammatory mediators, including NO, iNOS and TNF-α, in LPS-stimulated BV-2 cells Keywords : Suaeda asparagoides, DPPH, Anti-inflammatory activity, Microglial cells, iNOS, TNF-α

Study of Antibacterial Activity of Ethylacetate Total Extract and the Alkaloid Fraction from Flowering Aerial Parts of Glaucium vitellinum Boiss. et Buhse against Clinical Isolates of Staphylococcus Spp.

Background: The purpose of this study was evaluation of the antimicrobial effect of ethyl acetate extract and alkaloid fraction of Glaucium vitellinum (G. vitellinum) against clinical Staphylococcus spp. isolates from patients of Sina hospital of Tehran. M aterials and Methods: The plant of G. vitellinum was collected from Khonsar, Isfahan province, during May 2014. It’s flowering and aerial parts were washed, dried, powdered and extracted with methanol and ethyl acetate by using percolator apparatus, separately. In continuation, the alkaloid fraction was separated from metanolic total extract. 100 clinical isolates of Staphylococcus spp . were collected randomly from different clinical samples of patients who referred to Sina hospital of Tehran during 2013-2014. Also, their resistant to common antibiotics were evaluated by disk diffusion method based on the CLSI 2014 protocol. Continuously, the antibacterial effect of ethyl acetate total extract and the alkaloid fraction against clinical isolates of Staphylococcus spp . were evaluated by determining the minimal inhibitory concentration (MIC) by microdilution method based on the CLSI 2014. Standard Staphylococcus aureus (PTCC1431) and Staphylococcus epidermidis (PTCC 1435) were evaluated, simultaneously. R es ults: Based on the results, 93% of isolates were coagulase positive and 7% were coagulase negative Staphylococci spp. (CoNS). All coagulase positive cocci were identified as Staphylococcus aureus and among 7% CoNS, 3% were identified as S. epidermidis , so, in the follow they are named only CoNS. The MIC of alkaloid fraction of G.vitellinum was: 17.87 mg/ml and 23.21 mg/ml against coagulase positive S.aureus and CoNS isolates , respectively . Also, MIC of ethyl acetate total extract of G.vitellinum was: 73.25 mg/ml and 98.21mg/ml on coagulase positive S. aureus and CoNS isolates, respectively . 61.29% of clinical S. aureus isolates were sensitive to ethyl acetate total extract and 100 % were sensitive to alkaloid fraction while 100% were penicillin resistant while only 60% of them were Trimethoprim/sulfamethoxazole (SXT) sensitive. Similarly, among CoNS isolates, 42.85% and 100% were sensitive to ethyl acetate total extract and alkaloid fraction, respectively. While 100% were penicillin resistant and only 42%were ciprofloxacin and doxycycline sensitivity. C onclusion: based on the existence of good antibacterial effect for alkaloid portion of G. vitellinum against clinical isolates of Staphylococcus spp. doing other in vitro and in vivo complementation tests are recommended for the further studies. Keyword: Glaucium vitellinum , Anti bacterial, Alkaloids, Papaveraceae

Anti-inflammatory activity of bartogenic acid containing fraction of fruits of Barringtonia racemosa Roxb. in acute and chronic animal models of inflammation

The fruits of Barringtonia racemosa are traditionally used in Indian medicine for the treatment of pain and inflammatory conditions. In this study, a fraction of ethyl acetate extract of fruits of B. racemosa (BREAF) was investigated for anti-inflammatory activity in experimental models of acute and chronic inflammation. Activity against acute inflammation was evaluated in inflammogens induced rat paw edema models. Whereas, effect in chronic inflammation was evaluated in cotton pellet granuloma and oxazolone induced delayed type hypersensitivity (DTH) model in mice. The BREAF exhibited dose dependent anti-inflammatory activity in both acute and chronic models at oral doses of 5, 10 and 20 mg/kg. BREAF inhibited both phases of carrageenan induced rat paw inflammation. The reduction in paw inflammation by BREAF was also evident in histamine and serotonin induced inflammation in rats. Effect of BREAF on DTH indicates inhibition of immune mediated inflammation. The reduction in cotton pellet granuloma by BREAF treatment shows inhibition of proliferative changes associated with chronic inflammation. Analysis of BREAF after chromatographic separations showed presence of bartogenic acid as a major constituent. Hence, it is proposed that anti-inflammatory effects of BREAF can be partially attributed to its bartogenic acid content. The minute doses at which this fraction shows anti-inflammatory effects emphasizes the need for further investigations on its efficacy in the immuno-inflammatory conditions.

Antibacterial and Antioxidant Activities of Acetogenins from Streptomyces sp. VE2; An Endophyte in Vernonia cinerea (L.) Less.

Strain VE2 was isolated from the stem tissue of Vernonia cinerea (L.) Less. and identified as Streptomyces sp. on the basis of morphology, chemotaxonomy and 16SrDNA sequencing. The fractionation of the crude ethyl acetate (CEA) extract from VE2 cultures led to the isolation of two acetogenins; squamocin and rollidecin B; these compounds and CEA extract had potential in antibacterial and antioxidant activities. The crude extract showed the highest activity against Salmonella Typhi ATCC19430 and Bacillus cereus ATCC7064, with MIC values of 32 µg/ml. Squamocin also showed the lowest MIC (32 µg/ml) and Minimum Bactericidal Concentration (MBC) (128 µg/ml) against S. Typhi and B. cereus with corresponding large diameter of the zone of inhibitions (27.5 and 28.2 mm, respectively). Rollidecin B showed the highest DPPH antioxidant activity with SC50 value of 58.92 µg/ml.

Cytotoxic and antibacterial naphthoquinones from an endophytic fungus, Cladosporium sp.

ObjectiveEndophytes have the potential to synthesize various bioactive secondary metabolites. The aim of the study was to find new cytotoxic and antibacterial metabolites from endophytic fungus, Cladosporium sp. isolated from the leaves of Rauwolfia serpentina (L.) Benth. ex Kurz. (Fam: Apocyanaceae).Materials and methodsThe endophytic fungus was grown on potato dextrose agar medium and extracted using ethyl acetate. Secondary metabolites were isolated by chromatographic separation and re-crystallization, and structures were confirmed by 1H NMR, 13C NMR and mass spectroscopic data. The cytotoxicity was determined by WST-1 assay and brine shrimp lethality bioassay, while antibacterial activity was assessed by disc diffusion method.ResultsTwo naphthoquinones, namely anhydrofusarubin (1) and methyl ether of fusarubin (2), were isolated from Cladosporium sp. The isolated compounds 1 and 2, by WST-1 assay against human leukemia cells (K-562) showed potential cytotoxicity with IC50 values of 3.97 μg/mL and 3.58 μg/mL, respectively. Initial screening of crude ethyl acetate extract and column fractions F-8 and F-10 exhibited noticeable cytotoxicity to brine shimp nauplii with LC50 values of 42.8, 1.2 and 2.1 μg/mL, respectively. Moreover, the isolated compound 2 (40 μg/disc) showed prominent activities against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Bacillus megaterium with an average zone of inhibition of 27 mm, 25 mm, 24 mm and 22 mm, respectively and the activities were compared with kanamycin (30 μg/disc).ConclusionOur findings indicate that anhydrofusarubin (1) and methyl ether of fusarubin (2) might be useful lead compounds to develop potential cytotoxic and antimicrobial drugs.

Physiological Active Substance Changes of Soybean Curd Residue Fermented by &lt;i&gt;Preussia aemulans&lt;/i&gt; and its Bioactivity

A new species of fungus Preussia aemulans isolated from the Cordyceps sinensis fruiting body was cultured by soybean curd residue using solid-state fermentation method. The content of polysaccharide, amino acid, protein, phenolics and nucleoside were massively improved under the optimum conditions, including 10% adding dosage of glucose, 3% adding dosage of beef extract, 15 days of fermentation time. The crude Polysaccharide (CPS) fraction indicated significant immunomodulatory activity on macrophage RAW 264.7 cells and the Ethyl Acetate Extract (EAE) fraction revealed the moderate anti-proliferation ability on human colon cancer cell DLD-1 at high concentration. Both of CPS and EAE showed excellent antioxidant activities properties. The results were demonstrated that the fermented Soybean Curd Residue (SCR) could be utilized as nontoxic, safe and functional food, food additive and feed.

Evaluation of Cytotoxicity, Carbohydrate, and Lipid Inhibitory Activity of Codonopsis lanceolata using Different Solvent Fractions

ABSTRACT This study was conducted to evaluate the cyto-toxicity and α-amylase, α-glucosidase, pancreatic lipase inhibition in vitro by different solvent fractions from the roots of Codonopsis lanceolata. The values of IC 50 against Calu-6 cell showed a high effect in n-hexane fraction (10.13 µg/mL) whereas DW fraction exhibited the weakest inhibition on cell viability, having an IC 50 value of over 1,000 µg/mL. The values of IC 50 against HCT-116 cell showed the highest activity in n-BuOH fraction (102.01 µg/mL), followed by n-hexane fraction (145.85 µg/mL), methylene chloride fraction (332.02 µg/mL), ethyl acetate fraction (462.93 µg/mL) and DW fracion (>1,000 µg/mL). α-Amylase inhibitory activity in methylene chloride fraction and ethyl acetate fraction was found to have a higher inhibitory effect with 24.5% and 25.6% than the other fractions. The highest α-glucosidase inhibitory activity was observed from the ethyl acetate fraction extract, while the extract of DW fraction showed the lowest level of inhibitory activity at given experiment concentration. The pancreatic lipase inhibitory activity of C. lanceolata was found to have a higher the effect in ethyl acetate fraction. Inhibition of lipase activity of the ethyl acetate fraction and n-hexane fraction showed a relatively high, while the extract of DW fraction showed the lowest level at given experiment concentration. These results suggested that the roots of C. lanceolata may assist in the potential biological activity on carbohydrate, lipid Inhibitory activity and anticancer activity.Keywords : Cytotoxicity, α-Amylase, α-Glucosidase, Pancreatic lipase, Codonopsis lanceolata

Avaliação do potencial antioxidante frente à oxidação lipídica e da toxicidade preliminar do extrato e frações obtidas das frondes de Dicksonia sellowiana (Presl.) Hook

RESUMOA espécie Dicksonia sellowiana, conhecida popularmente como xaxim, é uma samambaia natural do continente americano e encontrada no Brasil na Mata Atlântica. Em 2001 foi inserida na lista do IBAMA como espécie ameaçada de extinção em decorrência da exploração para a confecção de vasos para a jardinagem. O presente trabalho descreve o potencial antioxidante lipídico (TBARS e Sistema β-caroteno/ácido linoleico) do extrato bruto e frações obtidos através de aparato de Soxhlet de frondes de Dicksonia sellowiana, além da atividade citotóxica e hemolítica in vitro. Pelo método TBARS, todas as amostras testadas apresentaram atividade, destacando a fração acetato de etila e extrato bruto cuja atividade foi comparável ao padrão ácido ascórbico. No sistema β-caroteno/ácido linoleico, a fração acetato de etila e extrato bruto apresentaram inibição da oxidação do ácido linoleico, destaque para a fração acetato de etila que não se diferenciou estatisticamente do padrão BHT. Na avaliação da toxicidade preliminar, não fora observado atividade citotóxica e hemolítica do extrato bruto e frações nos modelos testados. Os resultados demonstram o potencial antioxidante da espécie vegetal nos modelos de inibição da oxidação lipídica sem apresentar toxicidade.