Antifungal activity in vitro of Baccharis glutinosa and Ambrosia confertiflora extracts on Aspergillus flavus, Aspergillus parasiticus and Fusarium verticillioides

The aims of this study were to evaluate the antifungal properties of Baccharis glutinosa and Ambrosia confertiflora extracts against Aspergillus flavus, A. parasiticus and Fusarium verticillioides, and to isolate the group of compounds that are responsible for the antifungal activity. Samples of aerial parts from each plant were extracted with 70% methanol and sequentially partitioned with hexane, ethyl acetate, and n-butanol. The partitioned fractions were evaluated in their capacity to inhibit the radial growth of the three species of fungi. The active fraction was used for an assay-guided chromatography of antifungal extracts. The results showed that the extract from B. glutinosa partitioned in ethyl acetate (Bea) showed the highest antifungal activity against the three fungi. Bea completely inhibited the growth of F. verticillioides at 0.8 mg/ml, whereas the radial growth of A. flavus and A. parasiticus was inhibited 70% at 1.5 mg/ml. The purified antifungal fraction from Bea showed 72, 54, and 52% of antifungal activity, respectively.     

Biological activity of maize leaf extracts against the African armyworm,Spodoptera exempta

Maize (Zea mays L.) leaf tissue of cv Bastille and cv Michoacan 12 was extracted with n-hexane. The extracts were bioassayed against 5th instar African armyworm,Spodoptera exempta (Walker)(Lepidoptera: Noctuidae), by feeding the larvae on agar based media or sucrose impregnated glass fibre discs. The hexane extract of the ‘resistant’ cv Bastille exhibited feeding deterrency and toxicity which were not shown by the ‘susceptible’ cv Michoacan 12. The hexane extract of cv Bastille was adsorbed onto silica gel, the solution filtered off and the adsorbed component taken up into ethyl acetate. Bioassay of these fractions indicated that the toxic and deterrent action was retained in the ethyl acetate fraction. Preparative thin layer chromatography of the ethyl acetate fraction isolated two biologically active constituents. These were both growth inhibitors and lethal by ingestion to the 5th instar African armyworm. Implications for resistance in maize varieties to insect pests are discussed.     

Changes of amino acid composition and protein technical functionality of edible insects by extracting steps

Interest in edible insects is increasing with increase population and food demand, and research on their applications is underway. Quality changes of protein during extraction from edible insects are crucial for food application. Therefore, we investigated the amino acid composition and protein technical functionality extracted from three different edible insects (Tenebrio molitor, Allomyrina dichotoma, Protaetia brevitarsis). Three steps used for protein extraction were: grounding, de-fatting, and extraction. Essential amino acid index was increased undergone protein extraction steps. Allomyrina dichotoma had the best value at all steps and Protaetia brevitarsis was followed. The pH values of the extract increased with each step except for Tenebrio molitor. Further, lightness, redness, and yellowness decreased generally. Protein molecular weight differed according to species and Protaetia brevitarsis had the widest protein bands after extracting. Through extraction steps, foam capacity and stability were improved and emulsion capacity and stability did not improve dramatically. In conclusion, Protaetia brevitarsis was the most suitable edible insect to use for protein sources and protein extraction can be useful way to improve protein technical functionality of edible insects.     

Mortality of second-instar larvae of Tuta absoluta produced by the hexane extract of Lycopersicon hirsutum f. glabratum (PI 134417) leaves

1 The mortality resulting from exposure of second‐instar larvae of Tuta absoluta to a hexane extract of Lycopersicon hirsutum f. glabratum (PI 134417) leaves was evaluated in the laboratory. 2 A crude hexane extract was fractionated on a silica gel column to produce 17 fractions. The toxicity of the fractions was evaluated on T. absoluta larvae at 2 and/or 6 mg/10 larvae/Petri dish along with a control. 3 Only two fractions, 4.50 and 10.18% of the total hexane extract, were found to result in very high mortality of second‐instar T. absoluta. 4 All the 17 fractions were analysed by capillary gas chromatography/mass spectrometry (GC/MS). The two most toxic fractions were virtually identical and contained mainly undeca‐2‐one (2‐UD) and trideca‐2‐one (2‐TD). 5 A third major sub‐fraction 13 (50.73% of the total hexane extract) was further fractionated on a silica gel column to obtain five additional sub‐fractions. The toxicity of all the five sub‐fractions was evaluated using second‐instar T. absoluta at 2 mg/10 larvae/Petri dish and also analysed by GC/MS. The most toxic sub‐fractions contained mostly 2‐UD and 2‐TD. 6 The results suggest that 2‐UD and 2‐TD could be related to resistance of L. hirsutum to T. absoluta.     

Characterization of Schinus lentiscifolius Marchand (Anacardiaceae) Bark Extract and Its Effects on Lymphocyte Oxidative Stress and Heat Shock Response

Schinus lentiscifolius Marchand has been used in folk medicine to treat immunoinflammatory related diseases, which are marked by OS and altered HSR. Our study aimed to evaluate OS and HSR in lymphocytes treated with S. lentiscifolius bark extracts. S. lentiscifolius barks were partitioned with solvents to obtain hexane (SL‐HEX), ethyl acetate (SL‐ACOET) and methanol (SL‐MEOH) extracts, and the presence of bioactive compounds was evaluated by thin layer chromatography. Total phenols were measured by the Folin–Ciocalteu method and flavonoids were identified by HPLC‐DAD‐ESI‐MS/MS. Antioxidant capacity was verified by DPPH method, cell viability by Trypan Blue method, lipid peroxidation by TBARS and HSP70 by immunoblotting. The SL‐ACOET extract presented higher content of phenolic compounds and antioxidant activity in vitro. It was able to reduce lipid peroxidation levels in lymphocytes induced by H₂O₂ and improved cell viability. The SL‐ACOET extract inhibited HSR by a decrease in both intracellular content and release of 70 kDa heat shock proteins (HSP70) and also by decrease extra‐to‐intracellular HSP70 ratio in lymphocytes submitted to heat shock (2 h, 41 °C). S. lentiscifolius bark extract has antioxidant activity and inhibitory effect on HSR probably due to the presence of polyphenols as the flavonoids quercetin and kaempferol.     

Costus spiralis (Jacq.) Roscoe: A Novel Source of Flavones with α‐Glycosidase Inhibitory Activity

Costus spiralis, a plant used in traditional Brazilian medicine for the treatment of complications in diabetes, was investigated. Assay of hexane, ethyl acetate, methanol, and aqueous fractions obtained by partition of a crude methanol extract of dried leaves of C. spiralis revealed that AGI activity was confined to the ethyl acetate fraction. Purification of this fraction yielded schaftoside and isoschaftoside. The AGI activities of the two flavones were lower than, but comparable with, that of the anti‐diabetic drug acarbose. In contrast, the IC₅₀ value of the ethyl acetate fraction was 1.95‐, 2.34‐, and 2.22‐fold higher than those of acarbose, schaftoside, and isoschaftoside, respectively. The results demonstrate for the first time that schaftoside and isoschaftoside are responsible, in part, for the AGI activity of C. spiralis. Our study suggests that further investigations into C. spiralis may lead to the discovery of additional compounds with antihyperglycemic activity.     

Bioefficacy of Ceasalpinea bonduc (L.) Roxb. against Spodoptera litura Fab. (Lepidoptera: Noctuidae)

Bioefficacy of hexane, chloroform and ethyl acetate extracts of the seeds of Ceasalpinea bonduc (L.) Roxb. was studied against third instar larvae of Spodoptera litura at 0.25, 0.5, 1, 2.5 and 5.0% concentrations. Significant antifeedant, larvicidal and pupicidal activities and least LC₅₀ values were observed in chloroform extract. The chloroform extract was subjected to fractionation using silica gel column chromatography. Six fractions were obtained; among these, the third fraction showed high antifeedant, larvicidal and pupicidal activities at 1000 ppm concentration. Abnormalities in adults were also observed. All the activities were concentration dependent. C. bonduc could be useful in integrated pest management programme.     

Feeding deterrence,larvicidal and haemolymph protein profiles of an Indian traditional medicinal plant Alangium salviifolium (L.F.) Wangerin on cluster caterpillar,Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae)

Different organic solvent crude extracts from the leaves of Alangium salviifolium (L.F.) Wangerin, were tested for their feeding deterrence, larvicidal activity and protein concentrations on the fourth instar larvae of Spodoptera litura L. under laboratory conditions. Treatments were given through two different host plant leaves such as brinjal and castor. The bioassay was carried out at different concentrations viz., 0.625, 1.25, 2.5 and 5%. The maximum antifeedant activity in brinjal (52.0%) and castor (29.97%) leaf discs was recorded in ethyl acetate extract as compared with hexane and chloroform extracts at 5% concentration. The result of larvicidal activity revealed that the maximum larval mortality was registered in brinjal leaves (65.81%) and castor with (57.48%) in ethyl acetate extract as compared to hexane and chloroform extracts at 5% concentration. The ethyl acetate extract of A. salviifolium contained alkaloids, diterpenoids, and saponins. The treatment also reduced haemolymph protein concentration after 48 h.     

Purification and Properties of Ferritin from the Last Instar Larval Hemolymph of Protaetia brevitarsis (Coleoptera)

Ferritin, an iron storage protein, has been purified from the last larval hemolymph of Protaetia brevitarsis (coleoptera) by KBr density gradient ultracentrifugation and resource Q (anion exchange chromatography) using fast performance liquid chromatography (FPLC) system. The iron content of ferritin was determined by atomic emission spectroscopy and FereneS stain. Ferritin of P. brevitarsis is shown to have two different subunits presented on a SDSPAGE in normal (N) and ironinjected (I) hemolymph. SDS PAGE revealed that the ferritin consists of two major polypeptides of 27 and 28 kDa in normal hemolymph. Interestingly, however, 30 kDa subunit was substituted for 28 kDa when iron was injected into the hemolymph. Apporximate isoelectric points of 27 kDa, 28 kDa, and 30 kDa ferritin subunits were 6.7, 6.75, and 6.8, respectively. Ferritin of P. brevitarsis was detected by FereneS stain and confirmed by Western blotting using its polyclonal antibody. Other characteristics such as amino acid composition and Nterminal amino acid sequence were investigated. Amino acid composition of ferritin (N and I) was rich in alanine, glycine, glutamine or glutamic acid and serine, but poor in histidine, arginine, methionine and phenylalanine.     

Determination of the Phytochemical Composition and Antioxidant Potential of Eugenia copacabanensis and Myrciaria tenella Leaves (Myrtaceae) Using a Saccharomyces cerevisiae Model

Eugenia copacabanensis and Myrciaria tenella are present in restingas of the Atlantic Forest, but little information is available about their chemical and biological potential. In this context, the hexane, dichloromethane, ethyl acetate and butanol fractions from the leaves of methanolic extract were analyzed by GC/MS and HPLC-DAD and the antioxidant potential was determined by DPPH and ABTS assays and using a Saccharomyces cerevisiae model. Dereplication allowed the identification of 68 compounds, 42 and 41 of which, respectively, are first reported here for E. copacabanensis and M. tenella. In vivo results revealed that the ethyl acetate and butanol fractions showed expressive antioxidant protection in the BY4741 and Δgsh1 strains, with greater impact on glutathione-deficient cells. With a high diversity of phenolic compounds, these polar fractions of E. copacabanensis and M. tenella leaves are potential protectors against intracellular oxidative stress.     

Phagodeterrence and insecticidal activity of Hyptis suaveolens (Poit.) against four important lepidopteran pests

This study was carried out to evaluate the antifeedant and larvicidal activities of Hyptis suaveolens Poit. (Lamiaceae) leaves crude extracts and their fractions against four lepidopteran pests namely Helicoverpa armigera (Hbn.), Spodoptera litura (Fab.), Earias vittella (Fab.) and Leucinodes orbonalis (G.). Hexane, chloroform and ethyl acetate extracts were tested by leaf disc and fruit disc no-choice methods at 1% and 1,000?ppm concentrations for crude and fractions, respectively. Ethyl acetate extract of H. suaveolens exhibited the maximum antifeedant and insecticidal activity at 1% concentration against all the tested insects. It was subjected to fractionation using silica column chromatography with different combinations of hexane and ethyl acetate used as mobile phase. Among the 15 fractions obtained, fraction 2 showed the maximum antifeedant and insecticidal activity against all tested insects at 1000?ppm concentration. Preliminary phytochemical analysis of fraction 2 showed the presence of terpenoids and alkaloids. H. suaveolens could be considered as a safe and eco-friendly insecticide for lepidopteron pest management.     

Phytochemical and Biological Activities of Pseudocalymma elegans: A False Garlic

Evaluation of phytochemical constituents and antioxidant and antimicrobial activities of hexane (PELH), dichloromethane (PELDCM), ethyl acetate (PELEA), and MeOH (PELM) extracts of young leaves of Pseudocalymma elegans have been carried out. Moreover, extracts have also been explored for the presence of sulphur containing compounds, 1,2‐dithiolane ( 33 ), diallyl disulfide ( 35 ), 3‐vinyl‐1,2‐dithiacyclohex‐5‐ene ( 37 ), and diallyl trisulfide ( 38 ) responsible for the garlic like smell of P. elegans. All the extracts were found to be antioxidant and showed potent inhibition with IC₅₀ values of 0.168 ± 0.001, 0.128 ± 0.002, 0.221 ± 0.011, and 0.054 ± 0.001, respectively, as compared to standard drugs ascorbic acid (AA) and butylated hydroxytoluene (BHT). The ethyl acetate extract (PELE) showed excellent activities against few Gram‐positive and Gram‐negative bacteria and some fungi as compared with standard drug ceftriaxone (3rd generation cephalosporin) and nystatin, respectively. Chemical constituents of hexane, dichloromethane, and ethyl acetate extracts were identified by gas chromatography‐mass spectrometry and mass spectral library search. Over all 55 chemical constituents were first time identified from the leaves which included branched and n‐hydrocarbons, fatty acids, fatty acid methyl esters, fatty alcohols, terpenes, alkaloid, vitamins, glycosides, aromatic compounds, and sulfur containing compounds. Two known chemical constituents, ursolic acid ( 1 ) and β‐amyrin ( 2 ), were also purified for the first time from the MeOH extract. To elucidate the structures of these compounds, UV, IR, EI‐MS, ¹H‐ and ¹³C‐NMR spectroscopy were used.     

Description of the Larva of <Emphasis Type="Italic">Protaetia famelica</Emphasis> (Coleoptera,Scarabaeoidea) from the Russian Far East

A previously unknown larva of Protaetia famelica is described and illustrated. The larva of P. famelica is similar to that of P. brevitarsis in the number of posterior epicranial setae and in the presence of anterior frontal setae but differs in the chaetotaxy of the head capsule, in the epipharynx structure, and in the number and position of spinules on the anal sternite. An original key to the known larvae of the genus Protaetia in the fauna of the Russian Far East is given.     

Enhancement of pheochromocytoma nerve cell growth by consecutive fractionization of <Emphasis Type="Italic">Angelica gigas</Emphasis> Nakai extracts

This work was to investigate the effect of flavonoids from Angelica gigas Nakai on the proliferation and differentiation of PC12 cells. Several solvents including hexane, chloroform, ethyl acetate, butanol and water consecutively partitioned. We determined the ethanol crude extract of Angelica gigas Nakai. The hexane fraction was shown to contain the highest number of flavonoids as follows; 21.48 mg/g and the composition of the flavonoids was as follows: 12.24 mg/g of quercetin, 4.39 mg/g of myricetin and 2.58 mg/g of catechin. In addition, this hexane fraction greatly increased both cell growth and outgrowth of the neurite, and whose effects were three times higher than those of the other fractions. The length of the neurites was measured as ca. 110 μm in adding 50 μg/mL of the hexane fraction, which was about the same as the case of adding 50 ng/mL of NGF as a positive control. This result indicates that the differentiation of PC12 cells by the addition of the hexane fraction was comparable to the case of adding NGF. The hexane fraction was also determined to prevent apoptosis of PC12 cells by suppressing DNA fragmentation. It is interesting that the mixture of three major flavonoids, quercetin, myricetin and catechin showed stronger activity on, both PC12 cell growth and neuritis outgrowth, than when adding each flavonoid alone. We believe this was due to the synergistic effects of the three flavonoids. The activities of these flavonoids from Angelica gigas Nakai are reported for the first time in this study.     

An ethyl acetate sensor utilizing cataluminescence on Y2O3 nanoparticles

A cataluminescence (CTL) sensor using Y₂O₃ nanoparticles as the sensing materials was proposed for the determination of ethyl acetate. This ethyl acetate sensor showed high sensitivity and specificity at the optimal temperature of 264°C. Quantitative analysis was performed at a wavelength of 425 nm. The linear ranges of CTL intensity vs ethyl acetate concentrations were 2.0–250 ppm (r = 0.9965) and 250–6500 ppm (r = 0.9997) with a detection limit (3σ) of 0.5 ppm. There was no response or weak response when foreign substances such as formic acid, n‐hexane, toluene, acetic acid, benzene, and formaldehyde passing through the surface of Y₂O₃ nanoparticles. The sensor had a long lifetime more than 80 h with 3600 ppm ethyl acetate. It had been applied successfully to determine ethyl acetate in artificial air samples. Copyright © 2009 John Wiley & Sons, Ltd.     

Caesalpinia palmeri: First Report on the Phenolic Compounds Profile,Antioxidant and Cytotoxicity Effect

This study aimed to determine the phenolic compounds profile, antioxidant potential and cytotoxicity of extracts and fractions of Caesalpinia palmeri. Methanolic extracts were generated from C. palmeri berries, stems and flowers. The latter was subjected to liquid-liquid partition, obtaining hexane, ethyl acetate and residues fractions. Results showed that the flower extract and ethyl acetate fraction had a larger concentration of phenolic compounds (148.9 and 307.9 mg GAE/g, respectively), being ellagic acid (6233.57 and 19550.08 μg/g, respectively), quercetin-3-β-glycoside (3023.85 and 8952.55 μg/g, respectively) and gallic acid (2212.98 and 8422.34 μg/g, respectively) the most abundant compounds. Flower extract and ethyl acetate fraction also presented the highest antioxidant capacity on all tested methods (DPPH, ABTS, ORAC and FRAP) and low cytotoxicity against ARPE-19 cells (IC₅₀ >170 μg/mL). C. palmeri possessed high antioxidant potential, associated with the presence of phenolic compounds and low cytotoxicity, suggesting that they could represent an option to counter oxidative stress.     

The social wasp Polybia fastidiosuscula Saussure (Hymenoptera: Vespidae) uses herbivore‐induced maize plant volatiles to locate its prey

Social wasps in the Polybia genus are important for use as pest‐control agents in agricultural systems. The objective of this study was to investigate the behavioural responses of Polybia fastidiosuscula Saussure (Hymenoptera: Vespidae) to volatiles from maize, both constitutive volatiles and those induced by the herbivory of Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae). To assess the behavioural response of P. fastidiosuscula to S. frugiperda larvae, undamaged plants, S. frugiperda‐damaged plants, mechanically damaged plants, mechanically damaged plants plus regurgitant from larvae and extracts from various treatments, bioassays were conducted in a Y‐olfactometer. In addition, the volatiles from plants subjected to different treatments were collected via aeration, and they were quantified and identified. The wasps showed a greater preference for plants with damage induced either by larval feeding or by being mechanically damaged plus regurgitant than for undamaged plants or either larvae alone or mechanically damaged plants. Wasps were more attracted to extracts from plants + S. frugiperda larvae and to an extract from mechanically damaged plants + the regurgitant of larvae compared to hexane. The primary compounds induced by herbivory for 5–6 h after the beginning of the damage or regurgitant treatment were identified as α‐pinene, β‐myrcene, (Z)‐3‐hexenyl acetate, limonene, (E)‐ocimene, linalool, DMNT, (E)‐β‐farnesene, TMTT and indole. The results presented here show that the social wasp P. fastidiosuscula uses herbivore‐induced plant volatiles from maize to locate its prey.     

Enantiomeric 9‐mer peptide analogs of protaetiamycine with bacterial cell selectivities and anti‐inflammatory activities

Protaetiamycine is an insect defensin, derived from the larvae of the beetle Protaetia brevitarsis. In our previous work, we designed 9‐mer peptide analogs of protaetiamycine, including 9Pbw2 (RLWLAIKRR‐NH₂), 9Pbw3 (RLWLAIWRR‐NH₂), and 9Pbw4 (RLWLAWKRR‐NH₂). 9Pbw2 and 9Pbw4 showed high antimicrobial activity without cytotoxicity, while 9Pbw3 with higher hydrophobicity compared to 9Pbw2 and 9Pbw4 showed high cytotoxicity as well as high antimicrobial activity (Shin et al., J. Pept. Sci. 2009; 15: 559–568). In this study, we investigated the anti‐inflammatory activities of 9Pbw2, 9Pbw3, and 9Pbw4 by quantitation of NO production in LPS‐stimulated RAW264.7 cells. The results showed that only 9Pbw3 has strong inhibition of NO production, implying that Trp⁷ as well as optimum level of hydrophobicity may play key roles in the anti‐inflammatory activity of 9Pbw3. In order to design potent anti‐inflammatory peptide with lower cytotoxicity as well as high stability from cleavage by protease compared to 9Pbw3, we synthesized 9Pbw3‐D , the all‐D ‐amino acid analog of 9Pbw3. 9Pbw3‐D showed less cytotoxicity against RAW264.7 cells as well as considerably stronger inhibition of NO production and inflammation‐induced cytokine production in LPS‐stimulated RAW264.7 cells than 9Pbw3. 9Pbw3‐D inhibited the gene expression of inflammatory‐induced cytokine significantly more than 9Pbw3 and showed high resistance to proteolytic digestion. Binding of 9Pbw3‐D with LPS caused higher enhancement of the FITC fluorescence as a result of its stronger interaction with LPS compared to that of 9Pbw3 and this result is in good agreement with their anti‐inflammatory activities. 9Pbw3‐D with higher anti‐inflammatory activity as well as lower cytotoxicity against mammalian cell compared to 9Pbw3 can be a potent noncytotoxic antibiotic candidates. Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd.     

Lentinula edodes (shiitake mushroom): An assessment of in vitro anti-atherosclerotic bio-functionality

Mushrooms have been highly regarded as possessing enormous nutritive and medicinal values. In the present study, we evaluated the anti-oxidative and anti-atherosclerotic potential of shiitake mushroom (Lentinula edodes) using its solvent–solvent partitioned fractions that consisted of methanol:dichloromethane (M:DCM), hexane (HEX), dichloromethane (DCM), ethyl acetate (EA) and aqueous residue (AQ). The hexane fraction (1 mg/mL) mostly scavenged (67.38%, IC₅₀ 0.55 mg/mL) the 2,2-diphenyl-1-picryl hydrazyl (DPPH) free radical, contained the highest reducing capacity (60.16 mg gallic acid equivalents/g fraction), and most potently inhibited lipid peroxidation (67.07%), low density lipo-protein oxidation and the activity of 3-hydroxy 3-methyl glutaryl co-enzyme A reductase (HMGR). GC–MS analyses of the hexane fraction identified α-tocopherol (vitamin E), oleic acid, linoleic acid, ergosterol and butyric acid as the bio-functional components present in L. edodes. Our findings suggest that L. edodes possesses anti-atherosclerotic bio-functionality that can be applied as functional food-based therapeutics against cardiovascular diseases.     

Effects of solvent fractions of Allomyrina dichotoma larvae through the inhibition of in vitro BACE1 and β‐amyloid(25–35)‐induced toxicity in rat pheochromocytoma PC12 cells

Amyloid‐β peptide (Aβ) generation initiated by β‐site amyloid precursor protein cleaving enzyme 1 BACE1 is a critical cause of Alzheimer's disease. In the course of our ongoing investigation of natural anti‐dementia resources, the ethyl acetate (EtOAc) fraction exerted strong BACE1‐specific inhibition with the half maximal inhibitory concentration (IC₅₀) value of 9.2 × 10^?5 μg/mL. Furthermore, Aβ(25–35)‐induced cell death was predominantly prevented by the EtOAc fraction of Allomyrina dichotoma larvae through diminishing of cellular oxidative stress and attenuating apoptosis by inhibiting caspase‐3 activity. Taken together, the present study demonstrated that A. dichotoma larvae possess novel neuroprotective properties not only via the selective and specific inhibition of BACE1 activity but also through the alleviation of Aβ(25–35)‐induced toxicity, which may raise the possibility of therapeutic application of A. dichotoma larvae for preventing and/or treating dementia.