Differential in vitro and in vivo responses of Akkermansia muciniphila to Odontosoria biflora (Kaulf.) C.Chr. [Lindsaeaceae] hexane extract in diet- and alloxan-induced BALB/c mice

Hexane, chloroform, ethyl acetate and methanol extracts of Hydnocarpus pentandra (Flacourtiaceae) seeds were tested for antifeedant, larvicidal, pupal mortality and adult deformations activities against Helicoverpa armigera. Crude extracts were screened at 0.5, 1.0, 1.5 and 2.0% concentrations. Bioassay-guided fractionation method was followed to isolate the active fraction from the crude extract. Active fraction was analysed by FT-IR, 1H NMR, 13C NMR and GC-MS. Hexane extract presented the highest antifeedant (87.89%), pupal mortality (41.67%) and adult malformation activities at 2% concentration. Seven different fractions were isolated from hexane extract, among which fraction-2 showed the highest antifeedant (81.43%) activity and recorded the lowest LC50 of 792.07 ppm. The fraction-2 contained two cyclopentenyl carboxylic acids, such as hydnocarpic acid (1) and chaulmoogric acid (2) in the ratio of 2:1. These compounds were major constituents in the active fraction of hexane extract of H. pentandra seeds. Fraction-2 can be used for agricultural pest management.

Diabetes mellitus is a metabolic disease characterized by hyperglycemia, with alterations in the metabolism of carbohydrates, lipids and proteins, where there is an absolute or relative deficiency in the secretion and/or action of insulin, or both. Currently, the complications of DM with oxidative stress have been related, causing oxidative damage to biomolecules that can not be counteracted by the defense antioxidant systems, since they are diminished. Such oxidative damage can be counteracted with the administration of antioxidants such as those present in the leaves of Justicia spicigera. Solvents of different polarity were used: Hexane, ethyl acetate and ethanol, since each solvent carries different types of compounds and were tested in vitro antioxidants of: Antilipoperoxidation, reducing power, reduction of phosphomolybdenum and reduction of DPPH radical. In the reduction of the radical DPPH, the hexane and ethanolic extracts equalized the reduction of ascorbic acid with 100% and the ethyl acetate extract 70%. For the reduction of phosphomolybdenum, the hexane extract reduced it by 28%, the extract of ethyl acetate by 94%, and the extract ethanolic by 100%. The reduction of Fe3+ in the hexane extract was 9%; 17% ethyl acetate extract and the ethanolic extract reduced it by 25%. In the antilipoperoxidation technique, the hexanic and ethanolic extract inhibited lipoperoxidation by 82 and 79% respectively, while the extract of ethyl acetate by 53%. For the in vivo tests, male Wistar rats were used to obtain the kidneys. The GSH/GSSG ratio of the kidney mitochondria, the diabetic control group and the hexane and ethyl acetate extracts was of 0.8, unlike the ethanolic extract that significantly improved it to 2.0. Since in DM there is alteration in lipid and protein metabolism, it was of interest to analyze cholesterol, triglycerides, urea and serum creatinine. In the serological tests, the hexanic extract presented only hypolipidemic effect in the diabetic group. Ethyl acetate and ethanolic extracts had hypoglycemic and hypotriglyceridemic effects in diabetic rats. In conclusión, the hexane extracts, ethyl acetate and ethanol from the leaves of Justicia, are of beneficial effects of a serological and antioxidant level.Support or Funding InformationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Antibacterial activity test and chemical component analysis of hexane, ethyl acetate and ethanol extract of Brotowali stem from Manokwari Regency against B. subtillis and E. coli have been carried out. The extraction of the T. crispa stems was carried out by the maceration method in stages based on the polarity of the solvent. Chemical component analysis was performed using GC-MS and phytochemical tests. Based on the identification of chemical compounds by phytochemical screening, it shows the presence of flavonoids in the hexane extract, while in the ethyl acetate and ethanol extracts there are alkaloids, flavonoids, saponins, and tannins. The results of the chemical component analysis of hexane, ethyl acetate and ethanol extracts with GC-MS showed that the hexane extract contained 55 chemical components with 11 main components, ethyl acetate extract consisted of 39 components with 8 main components and ethanol extract there were 57 components with 8 main components. The results of the antibacterial activity test using the well method showed that the hexane and ethyl acetate extracts showed better antibacterial activity than the ethanol extract against the two types of tested bacteria, namely E. coli and B. subtillis. The antibacterial activity of hexane and ethyl acetate extracts against E. coli was 11.00 and 14.00 mm, respectively and against B. subtillis were 7.00 and 7.50 mm, respectively.

The current research was designed to examine the antidiabetic and anti-microbial activities in the extracts of methanol, ethyl acetate and hexane from Flesh of Matoa (Pometia pinnata J. R. Forst. & G. Forst) to find out the best solvent to produce antidiabetic and anti-microbial active compounds. In the phytochemical screening, hexane extracted alkaloids and terpenoids, while ethyl acetate extracted saponin and methanol extracted tannin, phenol and terpenoids. In the antidiabetic activity by a-glucosidase inhibition methods, ethyl acetate showed the highest percentage of inhibition and the smallest value of IC50, followed by hexane and methanol compared to the Quercetin as the control group. The maximum percentage inhibition was 36.74 ± 0.07 for 100 µg/mL of concentration. The minimum value of IC50 was observed at 159.74 ± 0.65 µg/ml compared to the control group 6.04 ± 0.14 µg/ml. The smaller the IC50 value, the stronger the antidiabetic activity. Anti-microbial inhibition activities assessed using anti-microbial diffusion test method. The hexane and ethyl acetate extracts provide an inhibition of fungi and bacteria compared to Ketoconazole and Amoxicillin as the standard. The maximum fungi inhibition zone was observed at 4 mm for A. flavus and 9 mm for A. niger by Hexane extracts compared to the standard 14 and 24 mm. The maximum bacteria inhibition zone was observed at 7 mm for B. cereus by both of hexane and ethyl acetate extracts compared to the standard 5 mm. Compared to the anti-microbe inhibitory power categories, hexane and ethyl acetate was on the range of weak and moderate. The methanol extract could not inhibit all types of microbe. This study concluded that various extract of flesh of Matoa showed antidiabetic and anti-microbial activities at different value with varying potencies. The ethyl acetate extract was found to be more potent among all, followed by hexane and methanol.
 HIGHLIGHTS
 
 Phytochemical screening of Flesh of Matoa (Pometia pinnata R. Forst. & G. Forst) produced alkaloids, terpenoids, saponin, tannin, phenol and terpenoids
 In the antidiabetic activity by a-glucosidase inhibition methods, ethyl acetate showed the highest percentage of inhibition and the smallest value of IC50, followed by hexane and methanol
 The hexane and ethyl acetate extracts provide an inhibition of fungi and bacteria compared to Ketoconazole and Amoxicillin as the standard
 
 GRAPHICAL ABSTRACT

This study describes the effects of extracts and fractions of Persea willdenovii leaves against goat gastrointestinal nematodes and their cytotoxicity on Vero cells. The in vitro ovicidal and larvicidal activities of the crude ethanolic, hexane, ethyl acetate (EAE), butanolic and residual hydroethanolic extracts were assessed through the inhibition of egg hatching and larval motility assays. The most active extract (EAE) was then fractionated by chromatography in an open column containing silica gel, to furnish six fractions (Fr1-Fr6), which were also tested. The cytotoxicity of active extracts and fractions was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and trypan blue exclusion assay. The EAE and two fractions (Fr1 and Fr2) showed inhibitory activity in the egg hatching of gastrointestinal nematodes of goats in a concentration-dependent manner. The effective concentrations for 50% inhibition (EC50) of egg hatching were 2.3, 0.12 and 2.94 mg/ml for EAE, Fr1 and Fr2, respectively. All extracts and fractions were not effective in inhibiting 50% of motility of infective larvae. EAE and Fr2 had IC50 values (50% inhibitory concentration) of 4.95 and 2.66 mg/ml, respectively. Fr1 showed a slight cytotoxic effect (cellular inviability <30%) only after 48h of treatment (MTT test). Gas chromatography-mass spectrometry (GC-MS) analysis showed the presence of six fatty acid ethyl esters, a fatty acid methyl ester and a long-chain ketone in the most active fraction. These constituents identified in P. willdenovii can be related to the high ovicidal activity and relatively non-toxic effect of the extracts.

Polyherbal preparations have gained much attention as a potential source for discovering new drug therapeutics for the treatment of diabetes mellitus. The present investigation aims to determine in vitro and in vivo antidiabetic activity and the antioxidant potential of hexane, ethyl acetate and methanol extracts of a polyherbal mixture prepared from equal amounts of garlic cloves (Allium sativum L.), curry leaves (Murraya koenigii L. Sprengel), black pepper seeds (Piper nigrum L.) and rath goraka fruits (Garcinia quaesita Pierre). The standardization and in vitro antioxidant activity and antidiabetic activities were determined using standard methods. The in vivo acute antihyperglycemic activity of the hexane, ethyl acetate and methanol extracts was determined using an oral glucose tolerance test in streptozotocin-induced diabetic Wistar rats. Highest significant in vitro antioxidant capacity in terms of DPPH free radical scavenging (24.71 ± 0.01 ppm, p = 0.02) and reducing antioxidant power of ferric ion (23.29 ± 0.78 mol/dm3, p = 0.01) and in vitro antidiabetic properties in terms of α-amylase (25.74 ± 0.60 ppm, p = 0.01) and α-glucosidase (22.37 ± 0.06 ppm, p = 0.02) inhibition activities were observed in the hexane extract when compared with the respective standard compounds, ascorbic for antioxidant (DPPH 6.78 ± 0.03 ppm; FRAP 25.02 ± 0.21 mol/dm3) and acarbose for antidiabetic activity (α-amylase 5.68 ± 0.35 ppm; α-glucosidase 17.11 ± 0.62 ppm). In the glucose tolerance test, a significant improvement of glucose tolerance was found in the hexane (17.38%) and ethyl acetate (15.81%) extracts-treated groups at the therapeutic dose against the diabetic control group (p &lt; 0.05). The results obtained from the present evaluation showed that the hexane and ethyl acetate extracts of the polyherbal mixture could be considered as a potential source for developing antidiabetic agents targeting the management of diabetes mellitus.

Isolation, structure characterization and prediction of antioxidant activity of two new compounds from the leaves of Dodonaea viscosa native to the Sultanate of Oman

Numerous attempts have been made to control leptospirosis by using chemoprophylaxis, but with limited success. The present study was done to investigate the antileptospiral potential of hexane, ethyl acetate and methanol extracts of Zingiber zerumbet rhizomes. The extracts were assayed for antileptospiral activity using broth microdilution method against Leptospira interrogans (serovar Batavie, Canicola, Australis) and Leptospira biflexa serovar Patoc. The Z. zerumbet hexane extract exhibited antileptospiral activity, with IC50 values of 248 μg/mL against L. interrogans serovar Canicola, IC50 of 125 μg/mL against L. interrogans serovar Australis, IC50 of 15.63 μg/mL against L. interrogans serovar Batavie and IC50 of 109 μg/mL against L. biflexa serovar Patoc. However, both ethyl acetate and methanol extracts did not show any distinct antileptospiral activity. Since the hexane extract of Z. zerumbet showed antileptospiral activity, the DNA-damaging properties of this extract were tested according to their IC50 and IC25 values that were specific to each serovars. The DNA-damaging properties were determined by treating the selected Leptospira spp. with the hexane extract and subjecting its DNA to electrophoresis and analysis on agarose gels. The results demonstrated that the hexane extract had DNA-damaging properties towards L. biflexa serovar Patoc and L. interrogans serovar Australis, as proven by the appearance of fragmented DNA on the gels. We conclude that the Z. zerumbet hexane extract could inhibit the growth of Leptospira spp. serovar Patoc and Australis through DNA-damaging activity and thus, could be a potential antileptospiral agent. Further studies are needed to investigate the potential of this hexane extract as an antileptospiral agent using in vivo rat models of leptospirosis.

Leishmania amazonensis and Leishmania chagasi: In vitro leishmanicide activity of Virola surinamensis (rol.) warb

Diabetes mellitus is a major health problem in developed and developing countries. Costus speciosus (C. speciosus) is widely used in Indian medicine to treat various diseases including diabetes. Hexane, Ethyl acetate, Methanol, and aqueous crude extracts of C. speciosus administered to streptozotocin (STZ)-induced diabetic rats at the dose of 250, 400, 400, and 600 mg/kg, respectively, for 60 days. It was found that plasma glucose concentration was significantly (P < 0.05) decreased in all the extracts compared to control. Also hexane crude extract restored the altered tissue protein and pancreatic DNA, plasma insulin, and plasma C-peptide levels to near normal. Based on the results of the bio-assays, hexane extract fractionated on silica gel, and active principles have been isolated from the most active fractions. Thus, this study shows that C. speciosus hexane extract and its compounds have an antihyperglycemic action, and are able to ameliorate the diabetic state, and probably they can be used as an alternate therapy for diabetes.

Antinociceptive effect of 7-hydroxy-3,4-dihydrocadalin isolated from Heterotheca inuloides: Role of peripheral 5-HT 1 serotonergic receptors

The phytochemical composition and antimicrobial potential of extracts of fruit pulp of Landolphia owariensis was investigated. L. owariensis mesocarp flour was obtained by oven drying at 500C and pulverized while microwave assisted extraction was used to obtain hexane, chloroform, ethyl acetate and methanol extracts, which were concentrated in vacuo using microwave and thereafter air dried at ambient temperature. Antimicrobial sensitivity tests on fourteen (14) human pathogenic microorganisms were performed using diffusion method while minimum inhibitory concentration (MIC) and minimum bacterial/fungal concentration (MBC/MFC) tests were performed using dilution method. The oven dried L. owariensis fruit pulp flour contained 12.45±0.03 mg/100 g tannins and 6.39±2.78 mg/100 g total flavonoids but low levels of anthraquinones, alkaloids, cyanogenic glycosides, saponins, steroids and terpenes. The phytosterols consisted of cholesterol, cholestanol, campesterol, stigmasterol, beta sitosterol and tocopherol with stigmasterol being the most abundant (1.06±0.21 mg/100 g). The flavonoids constituents included orientin, isovitexin and keampferol. Microbiological analysis of hexane, chloroform, ethyl acetate and methanol extracts of L. owariensis fruit pulp showed antimicrobial activities against S. aureus, S. pyogenes, S. typhi, S. dysenteriae, K.pneumoniea, C. albicans, C. krusei, C. stellatoidea and M. rubrum. Chloroform extract exhibited the highest zones of inhibition, followed by hexane and ethyl acetate extracts while methanol extract had least diameter of zones of inhibition. Also ethyl acetate extract was more potent (2.5-5 mg/mL), followed by chloroform and methanol extracts while hexane extract had the least potency (5-10 mg/mL) level. Similarly, ethyl acetate extract was most effective (5-10 mg/mL) in inhibiting bacterial/fungal growth. L. owariensis fruit pulp flour and extracts contained important secondary metabolites that may account for its antimicrobial activities.Keywords: Landolphia owariensis, phytochemicals, anti-microbial activities, flavonoids, phytosterols

Differential Inhibition of Seed Germination by Sweetpotato (<i>Ipomoea batatas</i>) Root Periderm Extracts

Mosquito larvicidal and ovicidal properties of Eclipta alba (L.) Hassk (Asteraceae) against chikungunya vector, Aedes aegypti (Linn.) (Diptera: Culicidae)

The feeding deterrent properties of hexane, ethyl acetate, and butanol lipophilic extracts of 16 species of Bermudian sponges presented to the sergeant major,Abudefduf saxatilus, and the Bermuda bream,Diplodus bermudensis, were investigated in the laboratory. The sponges investigated spanned a variety of different color morphs and were representative of patch reef, mangrove, rock wall, soft bottom, seagrass, and cave habitats. Six of the 16 sponges (36%) had at least one extract that deterred feeding byA. saxatilus, while only one of 16 sponges (6%) had an extract that deterred feeding byD. bermudensis. Among those sponges that deterred feeding in one or both fish, deterrence was most frequently associated with compounds extracted in ethyl acetate (moderately polar). Butanol extracts (most polar) were active in only half as many instances, while in only one case was a hexane extract (least polar) active. Both brighly colored (yellow, orange–yellow, gray–blue, purple–brown) and weakly colored (dull brown, pinkish white) sponges had hexane, ethyl acetate, or butanol extracts that were deterrent. Generally the numbers of sponges with bioactive lipophilic extracts that caused feeding deterrence in fish was low. This may be related to Bermuda being located just within or beyond the extreme northern range of many tropical fish and invertebrates, and therefore sponges are exposed to reduced levels of predation.