Methanol-soluble Fraction Research Articles

Bioassay Directed Fractionation of Petroleum Ether Extract of Aerial Parts of Ceriops tagal: Isolation of Lupeol as the Nematicidal Agent against Cyst Nematode Heterodera zeae.

Plant parasitic cyst nematode Heterodera zeae is a pest, causing substantial economic losses in agriculture. Organic pesticides, based on plant products have emerged as eco-benign nematicidal agents. Ceriops tagal is a well-known marine medicinal plant which has not been evaluated against any nematode. Petroleum ether extract of the aerial parts of the plant (CTP), exhibited promising activity against infective stage larvae of H. zeae. On subjecting to classical solvent-solvent separation, it afforded petroleum ether soluble (CTP-S), methanol soluble (CTPMS-1, CTPMS-2) and insoluble (CTPM-IN-2) fractions, which exhibited activity against the cyst nematode within 24 h exposure. GC, GC/MS and HR-ESI-MS analyses of CTPMS-1 and CTPMS-2 fractions resulted in the identification of a number of compounds, including pentacyclic triterpenoids, lupeol (1), betunal (2), betulin (3), lupenone (4), betulonaldehyde (5), betulonic acid (7), methyl 3-acetoxy-27-O-(3,4-dihydroxy-E-cinnamoyl)-20(29)-lupen-28-oate (8) and β-amyrin, along with phenylpropanoid esters, fatty acids and their derivatives, benzamide, and indole derivatives. CTPM-IN-2 which mainly contained lupeol (1) exhibited maximum nematicidal activity, with 91 % and 93 % mortality of the larvae of H. zeae, after exposure for 72 h at the concentration of 0.5 % and 1 %, respectively. Its fractionation and purification through column chromatography resulted in the isolation and identification of four lupane-type triterpenoids 1, 3, 4 and betulinic acid (6). One of its most abundant column fractions CC-9-18 (145 mg) which exhibited substantial activity, with 81 % mortality at the lowest concentration of 0.125 % after 48 h of incubation mainly contained lupeol. It seems lupeol, a wide spread bio-privileged triterpenoid is the nematicidal principle of the plant as its authentic sample showed LC50 value of 0.061 after 72 h exposure. It is for the first time that nematicidal activity is reported for any part of C. tagal and that of lupeol against H. zeae. Pentacyclic triterpenoids 1-8 are biosynthetically related. Of the twenty-four compounds isolated or identified in the present investigation only five constituents 1, 3, 6, 7 and palmitic acid have been isolated previously from C. tagal.

In vitro Analysis Provides New Insights into the Pharmacological Actions of Methanol Extract of Seeds of Tamarindus indica L. and its Kupchan Fractions

Tamarind or Tamarindus indica L. is a multipurpose plant distributed throughout the tropics including Bangladesh. The present study was conducted to establish the preliminary antioxidant, antimicrobial, anti-inflammatory and thrombolytic activities of methanol extract of T. indica seeds along with its Kupchan fractions. To evaluate the antioxidant properties, the total phenolic content of T. indica was determined and expressed in gallic acid equivalent (GAE). Alongside, DPPH free radical scavenging assay was performed to ensure the antioxidant properties of the seeds where the methanolic crude extract revealed the maximum activity having IC50 value of 9.43 μg/ml. In the antimicrobial assay by disk diffusion method, only non-polar fractions of the extract showed mild antimicrobial activity against the test organisms tested while the polar crude methanol extract exhibited the maximum inhibition (58.16%, p < 0.001) of hypotonic solution-induced erythrocyte rupture in anti-inflammatory investigation among all the partitionates. During evaluation of thrombolytic activity in terms of percent of clot lysis, the methanol soluble fraction exhibited the highest percent of thrombolysis (23.5%) as compared to the reference standard, streptokinase (64.25%). The findings of the current study rationalize some of the traditional uses of T. indica and preliminarily ascertain its bioactive potential, which may act as a base for phytochemical and mechanism-based pharmacological studies of the plant in future. Bangladesh Pharmaceutical Journal 25(1): 9-15, 2022 (January)

Antioxidant, Analgesic and CNS Depressant Activities of Commelina diffusa Burm. f.

Commelina diffusa Burm. f. is a member of Commelinaceae family, which is widely grown in crop land and treated as a weed. This plant has several important medicinal properties which have not been studied extensively. In this study, the crude powder of C. diffusa whole plant was extracted with 95% ethanol and different solvent fractions (n-hexane, chloroform and methanol) were prepared from the crude extract by solvent-solvent partitioning. All these plant samples were subjected to bioassays for evaluating the antioxidant, central analgesic, peripheral analgesic and CNS depressant activities. The crude extract and its methanol soluble fraction showed mild free DPPH scavenging property with IC50 values of 98.49- and 84.77-μg/ml, respectively as compared to the standard ascorbic acid (IC50 = 2.67 μg/ml). In the analgesic activity test, the n-hexane fraction of C. diffusa at doses of 200- and 400-mg/kg body weight exhibited significant (p<0.05) central analgesic activity (tail flick test) in mice. Similarly, all the test samples showed statistically significant (p<0.05) reduction in abdominal writhing induced by acetic acid. C. diffusa showed significant CNS depressant activity which was measured by ‘open field test’ and ‘hole cross test’. Considering the potential bioactivities, the plant materials can be further studied elaborately to explore the activities of the purified compounds to aid in future drug development. Dhaka Univ. J. Pharm. Sci. 20(2): 159-166, 2021 (December)

ISOLATION AND IDENTIFICATION OF A NEW ANTIPROLIFERATIVE INDOLOCARBAZOLE ALKALOID DERIVATIVE EXTRACTED FROM FARMED SHRIMP (Litopenaeus vannamei) MUSCLE

Farmed shrimp Litopenaeus vannamei, one of the most consumed seafood in the planet, is a source of antiproliferative extracts still to be fully characterized. This study the isolation and identification of these antiproliferative compounds. From a chloroform-soluble extract from shrimp muscle, hexane- and methanol-soluble fractions were obtained and tested for antiproliferative activity (MTT), a bioassay that guided the fractionation and isolation of bioactive fractions using open column chromatography. MeOH-soluble fraction resulted bioactive and was subjected to further fractionation from which one subfraction outstand for being highly active against prostate cancer cell line. Antiproliferative effects were evaluated using colorimetric assays and cell morphology observations. Further chromatographic procedures resulted in sub-fractions from which one was effective in causing DNA damage and F-actin polymerization, which suggests cellular collapse and apoptosis. According to the structural chemical characterization carried out, dioctyl phthalate, eicosapentaenoic acid, and an indolocarbazole alkaloid type of compound were identified. This last compound, which resulted majorly responsible for the bioactivity, was not found reported in the available databases. Pure EPA control was used to compare it with the subfraction, observing greater activity in the subfraction than when EPA was used, suggesting that another compound different from EPA is providing the highest activity; but, more investigation is needed for a full chemical and structural characterization.

Isolation and Characterization of Antimicrobial Compound from Stem Bark of Traditional Medicinal Plant Sonneratia apetala: Evaluation of their antimicrobial, Cytotoxic and Antioxidant Properties

Sonneratia apetala (S. apetala) (Lythraceae) has been investigated for the isolation and characterization of antimicrobial compounds and evaluation of their biological activities. The chloroform extract of the stem bark and different partitionate of the chloroform extracts i.e. Petroleum ether soluble fraction (PESF), Ethyl acetate soluble fraction(EASF), Methanol soluble fraction(MSF) and aqueous soluble fractions (ASF) were subjected to different chromatographic techniques to isolate secondary metabolites. Successive chromatographic separation and purification yielded a total of two compounds identified and characterized as Taraxerone(1) and 5,8-dihydroxy- 6-methoxy-4,9-dioxo-1,3,4,9-tetrahydronaphthol[2,3-c]furan-1-yl acetate (2) by extensive proton NMR spectrum (1H-NMRspectrum) analysis. The different partitionate like PESF, EASF, MESF and ASF were subjected to screen their antimicrobial properties against some selected Gram positive and Gram negative bacteria and fungi, brine shrimp lethality and antioxidant activities. The maximum zone of inhibition of chloroform extract was found against Pseudomonas sp. (16mm). All fractions showed more activity against Gram negative bacteria then Gram positive bacteria. In the brine shrimp lethality bioassay, among all extracts, the petroleum ether and ethyl acetate soluble fraction showed significant lethality having the LC50 value of 7.72 μg/ml. The antioxidant activity was evaluated in terms of determination of free radical scavenging activity (DPPH assay). Among all the extracts of S. apetala the highest free radical scavenging activity showed by (Methanol soluble fraction) MESF with IC50 value 18.0 μg/ml.
 Bangladesh J Microbiol, Volume 38, Number 1, June 2021, pp 1-5

Measurement report: Long-emission-wavelength chromophores dominate the light absorption of brown carbon in aerosols over Bangkok: impact from biomass burning

Abstract. Chromophores represent an important portion of light-absorbing species, i.e., brown carbon. Yet knowledge of what and how chromophores contribute to aerosol light absorption is still sparse. To address this problem, we examined soluble independent chromophores in a set of year-round aerosol samples from Bangkok. The water-soluble fluorescent chromophores identified via excitation–emission matrix (EEM) spectroscopy and follow-up parallel factor analysis could be mainly assigned as humic-like substances and protein-like substances, which differed in their EEM pattern from that of the methanol-soluble fraction. The emission wavelength of fluorescent chromophores in environmental samples tended to increase compared with that of the primary combustion emission, which could be attributed to secondary formation or the aging process. Fluorescent indices inferred that these light-absorbing chromophores were not significantly humified and comprised a mixture of organic matter of terrestrial and microbial origin, which exhibited a different characteristic from primary biomass burning and coal-combustion results. A multiple linear regression analysis revealed that larger fluorescent chromophores that were oxygen-rich and highly aromatic with high molecular weights were the key contributors of light absorption, preferably at longer emission wavelengths (λmax⁡ > 500 nm). Positive matrix factorization analysis further suggested that up to 50 % of these responsible chromophores originated from biomass burning emissions.

Hepatoprotective activity against acetaminophen-induced liver dysfunction and GC-MS profiling of a brown algae Sargassum ilicifolium

BackgroundDrug-induced hepatotoxicity is one of the most important causes of liver dysfunction. Acetaminophen (paracetamol) an analgesic-antipyretic drug is generally considered safe but its overdose may cause liver toxicity. Marine macro-algae (seaweeds) especially brown seaweeds possess unique biological activities including hepatoprotective potential. The current study focused on the hepatoprotective effect of different solvent fractions of Sargassum ilicifolium and characterization of its n-hexane soluble fraction.MethodsThe ethanol extract (20 g) of S. ilicifolium was mixed with solvents of increasing polarity, starting with n-hexane followed by chloroform and methanol. All three (n-hexane, chloroform and methanol) soluble fractions were administered to the rats at dose of 150 mg/kg, b.w. Intraperitoneal administration of acetaminophen (600 mg/kg b.w.) to rats was used to cause liver injury. The hepatic damage was evaluated by liver markers enzymes; aspartate aminotransferases (AST), alanine aminotransferases (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), bilirubin along with other metabolites i.e., triglycerides, cholesterol, urea, glucose and creatinine. Lipid peroxidation and glutathione and were estimated in liver tissue. n-Hexane fraction was subjected to GC-MS analysis in order to identify potent compounds.ResultsThe oral administration of n-hexane and methanol soluble fractions reduced the acetaminophen-augmented liver marker enzymes ALT, AST, ALP, LDH, along with bilirubin, urea, creatinine, glucose and triglycerides. The n-hexane and methanol soluble fractions also improved hepatic antioxidant level via enhancing hepatic glutathione and reversing lipid peroxidation. GC-MS spectroscopy of n-hexane fraction of S. ilicifolium revealed the presence of some new compounds. Among them, fatty acids were found to be in highest concentration followed by halogenated hydrocarbons, benzene derivatives, and sterols. Fatty acid in seaweed may be one of the factors for hepatoprotection from drug-induced hepatotoxicity.ConclusionFrom the results, it is evident that n-hexane and methanol soluble fractions of S. ilicifolium have the ability to protect the liver against toxicity, which is comparable with silymarin used as a standard drug. Sargassum ilicifolium contains bioactive compounds with pharmaceutical importance.

Increased Reproductive Capacity of Sprague Dawley Male Rats Assessed from the Number of Leydig Cells, Sertoli Cells, Primary Spermatocytes, and the Diameter of The Seminiferous Tubules through the Effect of Methanol Extract, Soluble and Insoluble Fractions Of N-Hexan Parijoto Fruit (Medinilla Speciosa Blume)

Increased Reproductive Capacity of Sprague Dawley Male Rats Assessed from the Number of Leydig Cells, Sertoli Cells, Primary Spermatocytes, and the Diameter of The Seminiferous Tubules through the Effect of Methanol Extract, Soluble and Insoluble Fractions Of N-Hexan Parijoto Fruit (Medinilla Speciosa Blume)

Lemongrass (Cymbopogon citratus) Ethanolic Extract Exhibited Activities That Inhibit -glucosidase Enzymes and Postprandial Blood Glucose Elevation

Lemongrass is a common ingredient in Indonesian traditional herbal medicine that potently inhibits carbohydrate hydrolysis. In this study, the in vitro α-glucosidase inhibitory (AGI) activity of lemongrass was compared with its in vivo activity to retard postprandial blood glucose elevation, and the bioactive compounds responsible for these activities were observed. Both water and ethanolic extracts of lemongrass (WLG and ELG, respectively) were tested in vitro for its inhibition of the sucrose and maltose hydrolyzing activities of rat intestinal glucosidase. ELG was observed to exert higher inhibitory activities (Sucrase IC50 = 8.74 mg/mL; Maltase IC50 = 18.93 mg/mL) than WLG. ELG was evaluated for its in vivo activity to retard blood glucose elevation in mice after sucrose, maltose, and glucose consumption. ELG was also fractionated using activity-guided chromatography, followed by liquid chromatography–mass spectrometry and high-performance liquid chromatography (HPLC). In vivo sugar tolerance test confirmed the AGI activity in a non-dose-dependent manner and showed potential additional mechanisms that may prevent postprandial hyperglycemia. The active principles were acquired in methanol-soluble fraction and purified using preparative thin-layer chromatography. HPLC analysis with commercial standards identified caffeic acid and kaempferol as the compounds responsible for the bioactivity of ELG. Results showed that lemongrass has the potential as herbal medicine ingredient in the management of diabetes.

Antioxidant, Thrombolytic, Antimicrobial and Cytotoxic Activities of Flavonoids Isolated from the Root Bark of Pongamia pinnata

The crude methanolic extract of the root bark of Pongamia pinnata was taken into consideration to isolate secondary metabolites. A total six known natural compounds were separated and purified by various chromatographic techniques and five isolates were identified as flavonoid derivatives such as pongachromene (1), kanugin (2), karanjin (3), demethoxykanugin (4), dimethoxypongapine (5) and the other is a triterpenoid (6). The pure compounds as well as petroleum ether, dichloromethane and ethyl acetate soluble fractions of crude methanolic extract were evaluated for bioactivities using established methods. In vitro antioxidant activity was studied by DPPH radical scavenging method using butylated hydroxyl anisole as standard. Among the pure compounds, kanugin and pongachromene showed significant antioxidant activity with the IC50 values of 27.20 ± 0.39 μg/mL and 43.53 ± 0.63 μg/ml, respectively as compared to the standard (23.87 ± 0.09 μg/ml), whereas karanjin, demethoxykanugin and dimethoxypongapine demonstrated moderate antioxidant activity. Mild thrombolytic activity was observed by different fractions with clot lysis ranging from 18.49 to 29.35% as compared to standard streptokinase (79.12%). The different solvent fractions and pure isolates showed very mild antimicrobial activity with zone of inhibition of 7.5 - 10.0 mm against the tested microorganisms using azithromycin and ketoconazole as standards. In the brine shrimp lethality bioassay, the dichloromethane, ethyl acetate, and methanol soluble fractions revealed significant lethality with LC50 values of 0.67 ± 0.05, 0.61 ± 0.13 and 0.56 ± 0.10 μg/ml, respectively as compared to standard tamoxifen (LC50 value 0.34 ± 0.09 μg/ml).
 Dhaka Univ. J. Pharm. Sci. 19(1): 1-8, 2020 (June)

Antidiabetic activity of Neolamarckia cadamba (Roxb.) Bosser flower extract in alloxan-induced diabetic rats

BackgroundVarious parts i.e., stem bark, leaf, root and flower of Neolamarckia cadamba (Roxb.) Bosser has been extensively used by traditional practitioners to treat all types of diabetes. Our present study investigated the antidiabetic property of the extract from its flower and evaluated its traditional use for diabetes.ResultsPhenolic contents were found to be higher in the methanol soluble fraction (MESF) (228.16 ± 0.32 GAE/g of sample) while flavonoids were greater in quantity in the chloroform soluble fraction (CSF) (291 ± 5.99 QE/g of sample). The MESF showed the highest free radical scavenging and α-amylase inhibitory potential with IC50 value of 30.52 ± 2.40 and 6.26 ± 0.82 μg/mL respectively and both the values were promising when compared to the individual standards. Highest reduction of blood glucose level were observed when the N. cadamba flower extract (NCFE) were given (not in combination with metformin) orally for 2 weeks. NMCE at 500 mg/kg resulted 60.2% (p < 0.05) reduction of blood glucose in the diabetic group, comparable with the effect of standard metformin (68.4%). Gain in body weight were also observed in the test groups indicating an overall improvement of health in the diabetic rats.ConclusionsThe extract of Neolamarckia cadamba (Roxb.) flower is rich in phenolic and flavonoid compounds attributing to its potent in vitro antioxidant and α-amylase inhibitory potential. Oral administration of NCFE also resulted in a significant (p < 0.05) dose dependent reduction of blood glucose level in diabetic rats, while no synergistic activity was observed when it was given in combination with metformin. These findings imply the possible pharmacological mechanisms behind the antidiabetic property of NCFE could be: inhibition of α-amylase and delaying the absorption of glucose from the intestine, secondly possessing ‘metformin like’ activity such as increasing cellular uptake and glycolysis of glucose, reducing gluconeogenesis in the liver etc. This study also for the first time provides scientific evidence in favor of the traditional use N. cadamba flower against diabetes and demands future study in order to isolate and characterize the bioactive compounds responsible for such bioactivities.

Methanol soluble fraction of fruits of Annona muricata possesses significant antidiarrheal activities

Medicinal plants are the major sources of traditional treatment of disease in Indian subcontinent due to abundant presence of plants and vast side effects of synthetic drug. The present study was subjected to observe in vitro thrombolytic, antibacterial, and in vivo antidiarrheal activities of methanol soluble fraction of fruits of Annona muricata. In thrombolytic activity assay, various concentrations (2 ─ 10 mg/ml) of methanol soluble fraction was used and dose dependently less potent activity was found. The maximum clot lysis 18.33% (p* < 0.05) was achieved at 10 mg/ml of methanolic fruit extract, whereas standard drug streptokinase showed 55.50% (p*** < 0.001) clot lysis. In antibacterial assay, disc diffusion method was used comprising two gram positive (S. aureus and Micrococcus luteus) and two gram negative (E. coli and P. aeruginosa) bacteria. None of four (0.25, 0.5, 1, and 5 mg/disc) concentration of fruit extract showed antibacterial potentiality, whereas standard amikacin (3 mg/disc) revealed strong antibacterial activities (=~ 23 ─ 24 mm of MIC). To evaluate antidiarrheal activity, castor oil induced diarrhea was created in Swiss albino mice and different doses (100, 200, and 400 mg/kgbw) of fruit extract was introduced post orally. All of three different doses of fruit extract showed significant (p < 0.05 ─ 0.001) antidiarrheal activities. Notably, the percent inhibition of diarrhea by methanolic extract of fruits of Annona muricata was found to be 58.38% at a dose of 400 mg/kgbw. The effect of vehicle saline (10 ml/kgbw) was considered as control and loperamide (5 mg/kgbw) as standard that provided 67.01% inhibition of diarrhea. The results suggest that, the fruits of Annona muricata possess potent antidiarrheal properties, providing scientific basis of using the plant parts in the treatment of diarrheal disease.

Dithiothreitol (DTT) activity of different fractions of fresh and ozonised soot and quantitative contributions of ozonised products of phenanthrene

The contributions of soluble and insoluble fractions of soot particles, as well as their ozonised products, to oxidative potential (OP), warrant investigation. In our study, soot particles generated under two air/fuel ratio (A/F) conditions were aged by ozone (O3) at an atmospherically relevant level in a chamber. The OP values of total soot particles and the methanol- and water-soluble fractions were measured by the dithiothreitol (DTT) assay (OPDTT). Results showed that both A/F and O3 aging influenced the OPDTT of different fractions. Lower A/F favoured increases in OPDTT for both total soot and its soluble fractions. The highest OPDTT increments after O3 aging were found in the methanol-soluble fraction of high-A/F soot and the insoluble fraction of low-A/F soot. Phenanthrene (PHE), one of the dominant polycyclic aromatic hydrocarbons (PAHs) in soot particles, was selected as a model PAH to investigate O3-PAH aging using in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), with phenanthrenequinone (PQ) and 1,1′-biphenyl-2,2′-dicarboxaldehyde (BD) identified as two major ozonised products. Furthermore, the quantitative contributions of PQ and BD to the OPDTT increases in ozonised soot were extrapolated. In particular, for ozonised low-A/F soot, PQ alone contributed 12.5% to the OPDTT increase. This work highlights the vital contributions of PQ and other key ozonised products of PAHs adsorbed on soot surfaces to the enhanced OPDTT values and potential toxicity of aged soot, which enhances current understanding regarding the health effects of aged soot in the atmosphere.

Antisickling and Antibacterial Activities of Anthocleista schweinfurthii Gilg. (Gentianaceae) from Non-human Primates Pharmacopoeia in Democratic Republic of the Congo

The aim of the present study was to evaluate the Chemical composition and bioactivity of Anthocleista schweinfurthii Gilg (Gentianaceae) fruits, leaves, root bark and stem bark extracts against Sickle cell disease (SCD) and associated bacteria. The antisickling and antibacterial activities were carried out using Emmel and micro-dilution methods respectively. The results revealed that the fruits, leaves, root bark and stem bark extracts of A. schweinfurthii contains various secondary metabolites such as the alkaloids, saponins, total polyphenols, flavonoids, tannins, anthocyanis, leuco-anthocyanins and quinones. The ethyl acetate and methanol extracts displayed antisickling activity. While, the antibacterial activity of different plant extracts tested was weak toward tested bacterial strains (CMI ≥125μg/mL). The antibacterial activity can be improved by bio-guided fractionation of the n-hexane, ethyl acetate or methanol soluble fraction. This study provides for the first time a scientific basis for the in vitro antisickling activity of A. schweinfurthii.

Antidiarrheal and Antimotility Activities of Stem Bark Extracts of Annona reticulata Linn. in Mice Model

The study was aimed to evaluate the phytochemical screening, in vivo evaluation of antidiarrheal activity, and GI motility of methanolic extract as well as different organic solvent soluble fractions of bark of Annona reticulata Linn. The powdered bark of the plant was extracted with methanol using cold extraction method and fractionated with solvent-solvent partitioning using organic solvents including n-hexane, chloroform and ethyl acetate. Phytochemical screening revealed the presence of alkaloids, flavonoids, phenolic compounds, diterpenes, carbohydrate, saponins, phenols, tannins and glycosides. The different organic solvent soluble fractions of bark were evaluated at a concentration of 200 mg/kgbw in castor oil induced diarrheal mice model. The aqueous soluble fractions of bark Annona reticulata showed highest percentage of inhibition of diarrhea (64.91 ± 1.37%), whereas methanol, n-hexane, chloroform and ethyl acetate soluble fraction showed 26.99 ± 1.79%, 34.85 ± 1.66%, 52.71 ± 1.42% and 45.45 ± 1.54% of diarrheal inhibition, respectively. At the same time, the reference standard Loperamide (5 mg/kg) exhibited 73.21 ± 2.06% inhibition of diarrhea. In GI motility test by charcoal plug method, the 200 mg/kgbw of aqueous soluble fraction showed highest antimotility activity (68.71 ± 3.98%), whereas methanol, n-hexane, chloroform and ethyl acetate soluble fractions showed 66.84 ± 3.38%, 52.01 ± 1.25%, 59.75 ± 3.56% and 54.70 ± 2.12% antimotility activity, respectively. The standard Loperamide (5 mg/kg) revealed 72.41 ± 1.33% inhibition of GI motility, whereas distilled water as control demonstrated 34.06 ± 1.09% of inhibition. This result indicates that the plant extracts have a significant inhibition of GI motility.

GPR31-dependent dendrite protrusion of intestinal CX3CR1+ cells by bacterial metabolites.

Small intestinal mononuclear cells that express CX3CR1 (CX3CR1+ cells) regulate immune responses1-5. CX3CR1+ cells take up luminal antigens by protruding their dendrites into the lumen1-4,6. However, it remains unclear how dendrite protrusion by CX3CR1+ cells is induced in the intestine. Here we show in mice that the bacterial metabolites pyruvic acid and lactic acid induce dendrite protrusion via GPR31 in CX3CR1+ cells. Mice that lack GPR31, which was highly and selectively expressed in intestinal CX3CR1+ cells, showed defective dendrite protrusions of CX3CR1+ cells in the small intestine. A methanol-soluble fraction of the small intestinal contents of specific-pathogen-free mice, but not germ-free mice, induced dendrite extension of intestinal CX3CR1+ cells in vitro. We purified a GPR31-activating fraction, and identified lactic acid. Both lactic acid and pyruvic acid induced dendrite extension of CX3CR1+ cells of wild-type mice, but not of Gpr31b-/- mice. Oral administration of lactate and pyruvate enhanced dendrite protrusion of CX3CR1+ cells in the small intestine of wild-type mice, but not in that of Gpr31b-/- mice. Furthermore, wild-type mice treated with lactate or pyruvate showed an enhanced immune response and high resistance to intestinal Salmonella infection. These findings demonstrate that lactate and pyruvate, which are produced in the intestinal lumen in a bacteria-dependent manner, contribute to enhanced immune responses by inducing GPR31-mediated dendrite protrusion of intestinal CX3CR1+ cells.

Aktivitas Antiplasmodium dan Sitotoksisitas Isolat Akar Pasak Bumi (Eurycoma longifolia Jack) secara In Vitro

Telah diketahui bahwa fraksi metanol akar pasak bumi mempunyai aktivitas antiplasmodium in vitro yang kuat. Namun belum diketahui isolat yang manakah yang aktif sebagai antiplasmodium. Penelitian ini bertujuan untuk menguji aktivitas antiplasmodium dan sitotoksisitas 5 isolat yang diisolasi dari fraksi metanol akar pasak bumi. Uji aktivitas antiplasmodium in vitro dilakukan pada kultur P. falciparum strain FCR-3 dan D10, sedangkan uji sitotoksisitas in vitro dilakukan pada kultur sel normal Vero. Aktivitas antiplasmodium dan sitotoksisitas in vitro dinyatakan sebagai IC50 (Inhibitory Concentration 50%) yaitu konsentrasi yang dapat menghambat pertumbuhan Plasmodium atau sel Vero sebesar 50%. Nilai IC50 ditentukan dengan analisis probit. Selektivitas isolat uji dinyatakan sebagai indeks selektivitas yang merupakan rasio sitotoksisitas (IC50 pada sel normal Vero) dengan aktivitas antiplasmodium in vitro (IC50 pada Plasmodium). Hasil penelitian menunjukkan bahwa isolat 4 dan 5 menunjukkan aktivitas antiplasmodium yang kuat dengan nilai IC50 berturut turut 2,21-19,02 dan 13,54-64,73 μg/mL Rasio sitotoksisitas/aktivitas antiplasmodium menunjukkan bahwa isolat 4 dan 5 juga menunjukkan nilai selektivitas yang tinggi yaitu 11.74-68.75 untuk isolat 4, dan 66.01-316.51 untuk isolat 5. Kesimpulannya, isolat 4 dan 5 mempunyai aktivitas aniplasmodium yang tinggi dengan seletivitas yang tinggi pula.

In Vitro Antimicrobial and Antioxidant Activities of Organic and Aqueous Extracts of Bacillus clausii KP10

Humans and plants are experiencing different kinds of diseases. Many of these diseases are mainly caused by microorganisms like fungi, bacteria and viruses that adversely affect their hosts. Bioactive compounds produced by bacteria can be used to treat the infections caused by these microorganisms. Free radicals that can cause serious damage to the cell can be repressed by antioxidants that are produced by bacteria as secondary compounds. The current case study was therefore planned to investigate the antimicrobial and antioxidant potentials of the organic and aqueous extracts of Bacillus clausii KP10 isolated recently by our group from hot springs of Tatta Pani, Poonch, Azad Kashmir. The antimicrobial activities were assessed using well diffusion method against two Gram positive bacteria; B. subtilis, and S. aureus, and two Gram negative bacteria; E. coli and P. multocida and three fungal strains; A. niger, A. parasiticus and A. ustus. Antioxidant activities of the extracts were done by DPPH free radical scavenging assay. In our experiments the maximum zones of inhibition were shown by ethanolic extract against P. multocida (21 mm), S. aureus (13 mm) and methanol soluble fraction showed zone of inhibition against A. ustus (21 mm) as compared to the standard ciprofloxacin for bacteria and as antifungal agents with terbinafine. In our studies n-hexane fraction of B. clausii KP10 showed maximum 88.67% antioxidant activity. Finally, results were analyzed statistically through analysis of variance (ANOVA). Hence it is proved that the bioactive fractions obtained from B. clausii KP10 are good source of antimicrobials and antioxidants and they can be used as medicines in future for the respective treatments.

Determination of Essential Oil Composition and Investigation of Phenolic Substance and Antioxidant Effect of “Helichrysum stoechas” from Hatay

This study was carried out to determine the essential oil composition, antioxidant capacity and totalphenolic substance of Helichrysum stoechas which was gathered from Hatay region. H. stoechas had a moisturecontent of 13.55 weight%, while the essential oil content of 0.3 wt% and ten volatile compounds were determinedand identified by GC. Predominantly volatile components are rosifoliol 4.89%, 3-methoxy-γ-asarone 34.69%,elemicin 3.24%, myristicin 23.35%, apiol 27.38%. Methanol soluble fraction was found to be 15.3%. Total phenolconcentration was found 256.55 ± 3.7 mg gallic acid equivalent/g of extract powder. DPPH free radical-scavengingactivity IC50 was found as 2.76 ± 44x10-2 mg mL-1. The inhibition value of methanolic extract was compared withBHA and BHT. The volatile oil is rich in hallucinogenic elemicin and myristicin. Obtained results showed that themethanolic extract was rich in phenolic content and has a close antioxidant activity to the synthetic antioxidantsBHT and BHA.

Cytotoxic activity and apoptosis induction of avocado Persea americana Mill. seed extract on MCF-7 cancer cell line

Avocado Persea Americana Mill. is a commercially important crop and studies have shown that the pulp may have benefits to cardiovascular health, dermatological health and possibly anti-cancer activity. Avocado seeds have several medicinal properties such as anti-hyperglycemic, antimicrobial, antioxidant and anti-inflammation. This study aim to evaluate the effect of avocado seed extract on viability and apoptosis of breast cancer cell line MCF-7. The anticancer effect was evaluated by cytotoxic test using MTT assay and the effect on apoptosis and cell cycle was examined by flow cytometry method. The cytotoxic test showed that chloroform extract had strong cytotoxic activity against MCF-7 cell lines with IC50 value of 94.87 µg/mL. Furthermore, the chloroform extract was partitioned with methanol and yield of soluble methanol fraction (FLM) and non soluble methanol fraction (FTLM). The cytotoxic activity of the methanol soluble fraction (FLM) and non soluble methanol fraction (FTLM) against MCF-7 cell lines was increased with IC50 of 34.52 and 66.03 µg/mL, respectively. Flow cytometry analysis using annexin-V and propidium iodide staining revealed that methanol soluble fraction could induce apoptosis and modulating the cell cycle arrest in MCF-7 cell. This research indicated that avocado seed has a potency to induce apoptosis and as anti-proliferative to MCF-7 cells lines.