Chemical composition and anti-fungal properties of the essential oils and crude extracts of Orthosiphon stamineus Benth

The hydrodistilled essential leaves and stems oils of Orthosiphon stamineus Benth were analysed by GC–MS/MS. Sixty nine compounds representing 97.6 and 97.4% of the total leaves and stems oils, respectively were identified, of which β-caryophyllene (24.0 and 35.1%), α-humulene (14.2 and 18.4%), β-elemene (11.1 and 8.5%), 1-octen-3-ol (8.2 and 7.0%), β-bourbonene (3.4 and 3.0%), β-pinene (2.1 and 1.7%), caryophyllene oxide (1.6 and 2.2%), camphene (1.6 and 1.3%) and limonene (1.2 and 1.1%) were the major compounds. Thus, the monoterpenes and sesquiterpenes were the predominant portions of the oils. Essential oils and methanol extract of O. stamineus and the derived fractions of hexane, chloroform, and ethyl acetate were tested for anti-fungal activity, which was determined by disc diffusion and minimum inhibitory concentration (MIC) determination methods. The oils, methanol extract and derived fractions of methanol extract displayed great potential of anti-fungal activity as a mycelial growth inhibitor against the tested phytopathogenic fungi such as Botrytis cinerea, Rhizoctonia solani, Fusarium solani, Colletotricum capsici and Phytophthora capsici, in the range of 49.3–70.3% and minimum inhibitory concentration ranging from 500 to 1000 μg/ml.     

α-Conotoxin TxID and [S9K]TxID, α3β4 nAChR Antagonists,Attenuate Expression and Reinstatement of Nicotine-Induced Conditioned Place Preference in Mice

Tobacco smoking has become a prominent health problem faced around the world. The α3β4 nicotinic acetylcholine receptor (nAChR) is strongly associated with nicotine reward and withdrawal symptom. α-Conotoxin TxID, cloned from Conus textile, is a strong α3β4 nAChR antagonist, which has weak inhibition activity of α6/α3β4 nAChR. Meanwhile, its analogue [S9K]TxID only inhibits α3β4 nAChR (IC₅₀ = 6.9 nM), and has no inhibitory activity to other nAChRs. The present experiment investigates the effect of α3β4 nAChR antagonists (TxID and [S9K]TxID) on the expression and reinstatement of nicotine-induced conditioned place preference (CPP) and explores the behaviors of acute nicotine in mice. The animal experimental results showed that TxID and [S9K] TxID could inhibit the expression and reinstatement of CPP, respectively. Moreover, both had no effect in acute nicotine experiment and the locomotor activity in mice. Therefore, these findings reveal that the α3β4 nAChR may be a potential target for anti-nicotine addiction treatment. [S9K]TxID, α3β4 nAChR antagonist, exhibit a superior effect for anti-nicotine addiction, which is promising to develop a novel smoking cessation drug.     

Effects of Cultivar and Environment on β-(1,3)-(1,4)- -glucan Content and Acid Extract Viscosity of Spanish Barleys

The acid extract viscosities and β-glucan contents of ten two- and six-rowed barley cultivars grown at seven locations in three consecutive years in Spain were studied in the present work. The viscosities varied from 2·4 to 24·8 centistokes (cSt) and the mean value was 6·4 cSt. The average β-glucan content of barleys determined by HPLC was 3·5% with a range of 1·9–5·5%. Significant differences were found in both β-glucan content and acid extract viscosity between different cultivars, locations and years. The β-glucan contents and viscosities of winter cultivars were higher than those of spring. Cvs. Barbarrosa and Hatif de Grignon were the genotypes with the highest values for both parameters, while cv. Beka had the lowest viscosity and β-glucan content. Environmental factors influenced both parameters. The acid extract viscosities of barleys were correlated negatively with the amount of precipitation (r=−0·754;P<0·05). Barleys grown in wet and rainy areas (Girona and La Coruña) had lower viscosity values.     

Comparative Analysis of Glycoside Hydrolases Activities from Phylogenetically Diverse Marine Bacteria of the Genus Arenibacter

A total of 16 marine strains belonging to the genus Arenibacter, recovered from diverse microbial communities associated with various marine habitats and collected from different locations, were evaluated in degradation of natural polysaccharides and chromogenic glycosides. Most strains were affiliated with five recognized species, and some presented three new species within the genus Arenibacter. No strains contained enzymes depolymerizing polysaccharides, but synthesized a wide spectrum of glycosidases. Highly active β-N-acetylglucosaminidases and α-N-acetylgalactosaminidases were the main glycosidases for all Arenibacter. The genes, encoding two new members of glycoside hydrolyses (GH) families, 20 and 109, were isolated and characterized from the genomes of Arenibacter latericius. Molecular genetic analysis using glycosidase-specific primers shows the absence of GH27 and GH36 genes. A sequence comparison with functionally-characterized GH20 and GH109 enzymes shows that both sequences are closest to the enzymes of chitinolytic bacteria Vibrio furnissii and Cellulomonas fimi of marine and terrestrial origin, as well as human pathogen Elisabethkingia meningoseptica and simbionts Akkermansia muciniphila, gut and non-gut Bacteroides, respectively. These results revealed that the genus Arenibacter is a highly taxonomic diverse group of microorganisms, which can participate in degradation of natural polymers in marine environments depending on their niche and habitat adaptations. They are new prospective candidates for biotechnological applications due to their production of unique glycosidases.     

Variability in essential oil and bioactive chiral monoterpenoid compositions of Indian oregano (Origanum vulgare L.) populations from northwestern Himalaya and their chemotaxonomy

The chemical composition of essential oils isolated from aerial parts of native populations of Origanum vulgare L. from northwestern Himalaya was investigated by enantioselective gas chromatography (enantio-GC), capillary-gas chromatography (capillary-GC) and GC–MS. Seventy compounds, which accounted for 84.9–97% of the oil composition, have been identified. The oils were allotted to two main groups; the first group had a high percentage of components belonging to the thymol/carvacrol biosynthetic pathway and the second was characterized by a different sesquiterpene composition and a high linalool and bornyl acetate percentage. Essential oils of O. vulgare populations from Rilkot and Dhanachuli areas in northwestern Himalaya were characterized by high monoterpenoids proportions as compared to Nainital and Bhowali populations, which were dominated by sesquiterpenoids. High proportions of thymol (29.2–82%) were found only in two populations. Composition of the samples collected from Dhanachuli area differed from Nainital and Bhowali collections by a unique presence of carvacrol. Notably, there were significant differences in the concentration of rest of the terpenes in all the collections. On enantio-GC, enantiomeric excess for (3R) (−)- and (3S) (+)-linalool, (S) (+)- and (R) (−)-terpinen-4-ol, (S) (−)- and (R) (+)-α-terpineol, (1S) (−)-bornyl acetate and (1S) (−)-borneol were established in populations from Bhowali and Dhanachuli area. Moreover, linalool type oregano may be useful in promoting wider commercial utilization of the biodiversity of this species.     

Optimization of air classification for the production of β-glucan-enriched barley flours

Barley grains contain a relevant amount of soluble fibre (i.e. β-glucans) and can be used for production of foods with healthy functions. To produce barley flours enriched in β-glucans, grain micronization and air classification of the flour was used, and a method to predict the relationship between the yield of the enriched flour and its β-glucan content was developed. This method is based on a series of sortings of micronized flour, with a progressive increase in the yield of a selected fraction; a curve for β-glucan enrichment vs. yield can then be calculated. Thus, the most suitable combination of yield and β-glucan content can be chosen. We tested this approach on two hull-less barleys with different starch type, and obtained barley flour fractions with twice the β-glucan concentration that the grain had. Enriched flour fractions with 11.2 and 15.6% β-glucans were obtained from cultivars Priora (hull-less) and CDC Alamo (waxy, hull-less), respectively, with a good flour yield (about 30%). We suggest that our method can be adopted for finely tuning the enrichment process to meet the industry's needs.     

Cereal β-glucans in diet and health

The native mixed linkage β-glucan of cereals is classified as a soluble dietary fibre, with rheological properties generally similar to guar gum and other random coil polysaccharides. The ability of oat and barley products to attenuate postprandial glycemic and insulinemic response is related to content of (1→3)(1→4)-β-d-glucan (β-glucan) and viscosity. A role of viscosity of β-glucan has not been directly demonstrated for lowering of serum cholesterol levels, and not all studies report a statistically significant lowering. The wide range of effectiveness reported may partially be explained by the properties of the β-glucan in the diets used, as well as the dose.     

Gelation of Waxy Barley Starch Hydrolysates

Thealpha-amylolysis of gelatinized waxy barley starch and the gelation of the hydrolysates was studied. The progress of the hydrolysis was monitored with gel permeation and ion exchange chromatography. Initial attack occurred mainly between the clusters, without significant hydrolysis of the external chains. The stiffness of 20% gels prepared from the polymeric products decreased with increasing hydrolysis of the amylopectin. A linear relationship between the shear modulus and the original high-molecular weight amylopectin content was observed. Although the branched degradation products of amylopectin (average DP <6×10⁴) were not involved in network formation, they retained their ability to recrystallize as analysed by differential scanning colorimetry. For more extensively hydrolysed products (average DP <5000) no recrystallization or gel formation was observed under the conditions tested. Information on the gel structure was obtained with acid hydrolysis and ion exchange chromatography. The structure of the acid-resistant gel residues was complicated, showing a five-peak distribution when analysed with ion exchange chromatography. No differences between the original and hydrolysate gels was observed. The gel residues were resistant to isoamylase.     

Alpha‐tocopherol and β‐carotene in legume–grass mixtures as influenced by wilting,ensiling and type of silage additive

Effects of wilting, ensiling and type of additive on α‐tocopherol and β‐carotene contents in legume–grass mixtures were examined. Swards of birdsfoot trefoil + timothy (Bft + Ti), red clover + timothy (Rc + Ti) and red clover + meadow fescue (Rc + Mf) were harvested as a first regrowth in August 2005. Forage was wilted to a dry‐matter (DM) content of 273 g kg^?1 and ensiled without additive or with an inoculant or acid. Wilting decreased α‐tocopherol concentration by 30% in the Bft + Ti mixture (P = 0·015). Untreated Bft + Ti silage had higher α‐tocopherol content than red clover silages (56·9 vs. 34·2 mg kg^?1 DM; P = 0·015). The α‐tocopherol concentration of Bft + Ti forages increased during ensiling from 41·1 mg kg^?1 DM in wilted herbage to 56·9, 65·2 and 56·8 mg kg^?1 DM in untreated, inoculated and acid‐treated silage respectively (P = 0·015). The inoculant increased α‐tocopherol content in the red clover silages (50·1 vs. 34·2 mg kg^?1 DM; P = 0·015) compared with untreated red clover silages. Red clover mixtures had lower β‐carotene content than Bft + Ti (32·3 vs. 46·2 mg kg^?1 DM; P = 0·016), averaged over treatments. In conclusion, wilting had small effects but the use of bacterial inoculant as an additive and a Bft + Ti mixture increased α‐tocopherol concentration in the silage.     

Changes in essential oil and fatty acid composition in coriander (Coriandrum sativum L.) leaves under saline conditions

The influence of salinity on essential oil and fatty acid composition of Tunisian coriander (Coriandrum sativum L.) leaves grown in hydroponic culture was investigated. Volatile constituents and fatty acid of leaves were analyzed. The essential oil yield increased significantly up to 18 and 43% with 25 and 50 mM NaCl, respectively and decreased significantly under high salinity. The major volatile compound in leaves was (E)-2-decenal with 52% of total essential oil constituents, followed by decanal, dodecanal, (E)-2-tridecenal and (E)-2-dodecenal. Further, the content of these compounds was affected differently by the treatment level.Salinity decreased significantly the total fatty acid content of the upper and basal leaves. α-Linolenic (C18:3n3) was the main compound of both organs, followed by linoleic (C18:2n6), heptadecenoïc (C17:1n7) and palmitic (C16:0) acids. Raising NaCl concentrations lead to an important decrease in the unsaturated to saturated fatty acid ratio stimulating the formation of more rigid membrane.     

Simulation of the factors affecting β-glucan levels during the cultivation of oats

A Monte Carlo simulation technique was employed to simulate the factors influencing the level of β-glucan content in both hulled (HO) and naked (NO) oat cultivars during cultivation of the crop. Probability density functions were used to model the uncertainty and variability in the input factors. A scenario analysis was subsequently developed to look at the impact of different model assumptions and input parameters. The simulated mean β-glucan level in harvested oats grain was 3.50 and 4.25 g/100 g for hulled (HO) and naked (NO), respectively. A sensitivity analysis highlighted that cultivar selection was the most important input parameter compared to other inputs in determining the final β-glucan level (correlation coefficients of 0.64 and 0.79 for HO and NO, respectively). The analysis also indicated the positive effect of delayed sowing on β-glucan content (correlation coefficients of 0.32 and 0.25 for HO and NO, respectively). Germination and storage factors showed a negative impact on the final β-glucan levels. The scenario analysis shows the applicability of the proposed model for various agronomic practices. This approach establishes a quantitative scientific ranking of factors influencing β-glucan levels during the cultivation of oats.     

Influence of growth temperature on content, viscosity and relative molecular weight of water-soluble β-glucans in barley (Hordeum vulgare L.)

β-Glucan content and viscosity of water-soluble β-glucans have a considerable impact on the digestion of barley. Eight different 2-row barley (Hordeum vulgare L.) cultivars were grown in controlled environment chambers at five different temperatures until maturity. The samples were pearled, milled and analysed for their content of total, insoluble and soluble β-glucan. The water-soluble fraction was extracted at 37 °C, subjected to amylase treatment and freeze-dried. Water-soluble crude β-glucans were solubilised in distilled water, analysed for viscosity and their purity was checked by nuclear magnetic resonance. Molecular weights were profiled by size exclusion chromatography with RI detection. The content of total β-glucan varied from 4.0% to 7.4%, and was significantly affected by the growth temperature. An interaction between cultivar and growth temperature was observed for the total β-glucan content. The extractability was significantly affected by growth temperature, as there was recorded an increasing amount of water-soluble β-glucan with increasing growth temperature. Both the viscosity and the molecular weight of the water-soluble β-glucans increased with the growth temperature.     

Cultivar and Environmental Effects on (1→3,1→4)-β-D-Glucan and Protein Content in Malting Barley

A low β-glucan and protein content is desirable for malting barley. In China high β-glucan content has been considered a major factor contributing to the inferior malting quality of local barleys. In this study 10 barley cultivars were planted in multi-location trials in the southern winter-barley zone to determine genotypic and environmental effects on β-glucan and protein content. There were highly significant differences in both β-glucan and protein content between the 10 barleys, the eight locations and the 2 planting years. The average β-glucan content over the 2 years for the 10 cultivars in the eight locations ranged from 3·91% for Gangpi 1 to 4·95% for Xiumei 3, and from 3·76% for Hangzhou to 4·75% for Taian in eight locations. Correspondingly, the protein contents of the 10 cultivars ranged from 11·37% for Yanyin 1 to 12·52% for ZAU, and of the eight locations from 9·5% for Hangzhou to 14·69% for Taian. Environmental factors contributed the largest component of the variation in both β-glucan and protein content. Five climatic factors, namely the total accumulated temperature, accumulated temperature to 25 °C and to 30 °C, precipitation and days with rain during seed development were included in a regression equation relating climatic factors and β-glucan content. In the regression equation relating climatic factors and protein content, two other factors, days from heading to maturity and accumulated temperature to 20 °C were included, in addition to the five climatic factors that significantly affected β-glucan content.     

The combined use of hull-less barley flour and xylanase as a strategy for wheat/hull-less barley flour breads with increased arabinoxylan and (1→3,1→4)-β-D-glucan levels

Bread-making with a composite flour (CF) consisting of 60% wheat flour (WF) and 40% hull-less barley flour, increased the total and soluble (1→3,1→4)-β-D-glucan and total arabinoxylan (AX) contents of dough and bread samples, but decreased the specific bread loaf volume. A xylanase insensitive to inhibition by Triticum aestivum L. xylanase inhibitor (TAXI) and xylanase inhibiting protein (XIP), increased loaf volume by 8.8 and 20.1% for WF and CF breads, respectively. Xylanase addition not only markedly improved loaf volume of CF bread, but also increased the soluble AX content of the WF and CF dough and bread samples because of conversion of water-unextractable AX into soluble AX. The xylanase had no impact on the extractability and molecular weight of (1→3,1→4)-β-D-glucan, but (1→3,1→4)-β-D-glucan was degraded during bread-making probably because of endogenous β-glucanase activity. Taken together, the results clearly show that the combined use of hull-less barley flour and a xylanase active during bread making, lead to palatable breads with high total and soluble AX and (1→3,1→4)-β-D-glucan contents. The sum of total AX and (1→3,1→4)-β-D-glucan was 1.70% for WF bread and 3.06% for CF bread, while the sum of soluble AX and (1→3,1→4)-β-D-glucan was 0.49 and 1.41% for control WF and CF xylanase supplemented breads, respectively.     

The Anticancer Effect of Fucoidan in PC-3 Prostate Cancer Cells

Fucoidan, a sulfated polysaccharide, has a variety of biological activities, such as anti-cancer, anti-angiogenic and anti-inflammatory. However, the mechanisms of action of fucoidan as an anti-cancer agent have not been fully elucidated. The present study examined the anti-cancer effect of fucoidan obtained from Undaria pinnatifida in PC-3 cells, human prostate cancer cells. Fucoidan induced the apoptosis of PC-3 cells by activating both intrinsic and extrinsic pathways. The induction of apoptosis was accompanied by the activation of extracellular signal-regulated kinase mitogen-activated protein kinase (ERK1/2 MAPK) and the inactivation of p38 MAPK and phosphatidylinositol 3-kinase (PI3K)/Akt. In addition, fucoidan also induced the up-regulation of p21^Cip1/Waf and down-regulation of E2F-1 cell-cycle-related proteins. Furthermore, in the Wnt/β-catenin pathway, fucoidan activated GSK-3β that resulted in the decrease of β-catenin level, followed by the decrease of c-myc and cyclin D1 expressions, target genes of β-catenin in PC-3 cells. These results suggested that fucoidan treatment could induce intrinsic and extrinsic apoptosis pathways via the activation of ERK1/2 MAPK, the inactivation of p38 MAPK and PI3K/Akt signaling pathway, and the down-regulation of Wnt/β-catenin signaling pathway in PC-3 prostate cancer cells. These data support that fucoidan might have potential for the treatment of prostate cancer.     

The Promoting Effect of Ishige sinicola on Hair Growth

This study was conducted to evaluate the promoting effect of Ishige sinicola, an alga native to Jeju Island, Korea, on hair growth. When vibrissa follicles were cultured in the presence of I. sinicola extract for 21 days, I. sinicola extract increased hair-fiber length. After topical application of I. sinicola extract onto the back of C57BL/6 mice, anagen progression of the hair shaft was induced. The I. sinicola extract significantly inhibited the activity of 5α-reductase. Treatment of immortalized vibrissa dermal papilla cells (DPCs) with I. sinicola extract resulted in increase of cell proliferation, which was accompanied by the increase of phospho-GSK3β level, β-catenin, Cyclin E and CDK2, whereas p27^kip1 was down-regulated. In particular, octaphlorethol A, an isolated component from the I. sinicola extract, inhibited the activity of 5α-reductase and increased the proliferation of DPCs. These results suggest that I. sinicola extract and octaphlorethol A, a principal of I. sinicola, have the potential to treat alopecia via the proliferation of DPCs followed by the activation of β-catenin pathway, and the 5α-reductase inhibition.     

Essential oil composition of Hypericum perfoliatum L. and Hypericum tomentosum L. growing wild in Tunisia

The essential oils obtained by hydrodistillation from the aerial parts of Tunisian native Hypericum perfoliatum L. (sect. Drosocarpium Spach.) and Hypericum tomentosum (sect. Adenosepalum Spach.) were analyzed by GC and GC–MS. Thirty-two compounds were identified in the essential oils of H. perfoliatum with α-pinene (13.1%), allo-aromadendrene (11.4%), germacrene-D (10.6%), n-octane (7.3%), α-selinene (6.5%) and β-selinene (5.5%) as main constituents. Sixty-seven components were identified in the oil of H. tomentosum with menthone (17.0%), n-octane (9.9%), β-caryophyllene (5.3%), α-pinene (5.2%), lauric acid (4.1%) and β-pinene (3.7%) as the most abundant components. Both oils were characterized by the presence of many components which could have numerous applications in food, pharmaceutical and perfume industries.     

First Identification of 12β-Deoxygonyautoxin 5 (12α-Gonyautoxinol 5) in the Cyanobacterium Dolichospermum circinale (TA04) and 12β-Deoxysaxitoxin (12α-Saxitoxinol) in D. circinale (TA04) and the Dinoflagellate Alexandrium pacificum (Group IV) (120518KureAC)

Saxitoxin and its analogues, paralytic shellfish toxins (PSTs), are potent and specific voltage-gated sodium channel blockers. These toxins are produced by some species of freshwater cyanobacteria and marine dinoflagellates. We previously identified several biosynthetic intermediates of PSTs, as well as new analogues, from such organisms and proposed the biosynthetic and metabolic pathways of PSTs. In this study, 12β-deoxygonyautoxin 5 (12α-gonyautoxinol 5 = gonyautoxin 5-12(R)-ol) was identified in the freshwater cyanobacterium, Dolichospermum circinale (TA04), and 12β-deoxysaxitoxin (12α-saxitoxinol = saxitoxin-12(R)-ol) was identified in the same cyanobacterium and in the marine dinoflagellate Alexandrium pacificum (Group IV) (120518KureAC) for the first time from natural sources. The authentic standards of these compounds and 12α-deoxygonyautoxin 5 (12β-gonyautoxinol 5 = gonyautoxin 5-12(S)-ol) were prepared by chemical derivatization from the major PSTs, C1/C2, produced in D. circinale (TA04). These standards were used to identify the deoxy analogues by comparing the retention times and MS/MS spectra using high-resolution LC-MS/MS. Biosynthetic or metabolic pathways for these analogues have also been proposed based on their structures. The identification of these compounds supports the α-oriented stereoselective oxidation at C12 in the biosynthetic pathway towards PSTs.     

Role of the Y134F mutation in cyp51 and overexpression of cyp51 in the sensitivity response of Puccinia triticina to epoxiconazole

A collection of 110 isolates of Puccinia triticina in 2007 from different European wheat-growing regions were examined for their sensitivity to the demethylation inhibiting (DMI) fungicide epoxiconazole and the expression and mutations of the target gene, cyp51. The cyp51 gene was highly conserved across Europe. A mutation at codon 134 (Y134F) was identified in only five isolates. This mutation had a limited impact, if any, on the sensitivity of P. triticina towards epoxiconazole. Over the entire collection of isolates, cyp51 expression and in vitro sensitivity towards epoxiconazole was moderately correlated. A constitutive up-regulation of the cyp51 gene was detected in some of the isolates with higher ED₅₀ values. Such up-regulation was not inducible by exposure to different concentrations of epoxiconazole. Isolates of P. triticina with the highest ED₅₀ values and with the Y134F mutation in the cyp51 gene or with cyp51 overexpression were equally well controlled in vivo by registered field rates of epoxiconazole as compared to an isolate with the lowest ED₅₀ value.     

Influence of environment on growing period and yield, protein, oil and α-linolenic content of three chia (Salvia hispanica L.) selections

Chia (Salvia hispanica L.) is an annual summer herb, and a member of the Labiatae family. Three selections, named Tzotzol, Iztac 1, and Tliltic, were planted at different elevations in three Inter-Andean valleys of Ecuador (Patate—2042 m, Guayllabamba—2200 m and Salinas—1621 m) to assess performance and seed quality. Length of growing period was influenced by location, with Tzotzol and Iztac 1 increasing with elevation. Iztac 1 had the highest seed yield at all three locations, with the difference between Iztac1 and Tzotzol being significant (P < 0.05). Similar protein contents were found among selections, within a site, except for Patate where Tliltic had a higher content. Seed at the Salinas site contained significantly (P < 0.05) higher α-linolenic and lower linoleic and oleic fatty acid contents than seed produced at the other two sites. In general seed yield, protein content, and fatty acid composition of the three selections were similar, and in some cases higher, than those reported for other ecosystems in which chia has been grown.