Cuticular lipids for species recognition of mole crickets (Orthoptera: Gryllotalpidae): II. Scapteriscus abbreviatus,S. acletus,S. vicinus,S. sp., and Neocurtilla hexadactyla

Qualitative analyses were made from whole-body cuticular extracts of Scapteriscus abbreviatus, Scapteriscus acletus, Scapteriscus vicinus, and Neocurtilla hexadactyla by isothermal and temperature-programmed gas chromatography. Adults of both sexes and nymphs of each species were collected in Florida. The chromatographic profiles of peaks were distinct and easily recognizable for each species, regardless of sex or developmental stage. Distinct sexual differences were found in S. acletus and S. abbreviatus. Specimens of S. abbreviatus from Puerto Rico and S. vicinus from Bolivia produced gas chromatography (GC) traces very similar to those of conspecifics collected in Florida. Evidence is presented to illustrate the potential importance of volatile cuticular lipid analysis as a tool for mole cricket identification. Cuticular extracts of an undescribed short-winged species of Scapteriscus from Bolivia were also examined and produced GC traces unlike those of any other species analyzed to date.     

Hydrocarbons in green and blue-green algae

Liquid column chromatography and thin-layer chromatography were used to determine the total content of hydrocarbons and gas chromatography was used to evaluate composition of hydrocarbons in green algae (Chlorella kessleri, C. vulgaris, Chlorella sp.,Scenedesmus acutus, S. acuminatus, S. obliquus) and the blue-green alga (Spirulina platensis) cultivated under autotrophic or heterotrophic conditions. InC. kessleri cultivated under heterotrophic conditions the content of hydrocarbons was found to be about 10^-2 % (per dry mass), whereas under autotrophic conditions it was about 10^-3 % (per dry mass). The highest content of hydrocarbons was detected in species of the genusScenedesmus cultivated autotrophically (10^-1 %). Heptadecane and hexacosane were found as major alkanes, 1-heptadecene was detected among alkenes.     

Comparative study of the yield of T-2 toxic produced by Fusarium poae,F. sporotrichioides and F. sporotrichioides var. tricinctum strains from different sources

Eleven strains of Fusarium poae and F. sporotrichioides from the U.S.S.R. and 7 strains of these species and one of F. sporotrichioides var. tricinctum from U.S.A. and France have been compared as to their capacity to yield T-2 toxin. The presence of this toxin was confirmed by thin layer chromatography, gas liquid chromatography and mass spectroscopy. The strains which came from the U.S.S.R. and were originally involved in causing Alimentary Toxic Aleukia (ATA), produced several times more T-2 toxin than the others. We assume that the ability of the U.S.S.R. strains to produce much more T-toxin is due to the different ecological conditions under which they were first isolated. The remarkable maintenance of so high a level of toxin production over more than 30 years of culturing is worthy of note.This work was partly supported by the Joint Research Fund of the Hebrew University and Hadassah.     

Isolation and characterization of gas vesicles from Microcyclus aquaticus

Intact gas vesicles of Microcyclus aquaticus S1 were isolated by using centrifugally accelerated flotation of vesicles and molecular sieve chromatography. Isolated gas vesicles were cylindrical organelles with biconical ends and measured 250×100 nm. The gas vesicle membrane was composed almost entirely of protein; neither lipid nor carbohydrate was detected, although one mole of phosphate per mole of protein was found. Amino acid analysis indicated that the protein contained 54.6% hydrophobic amino acid residues, lacked sulfur-containing amino acids, and had a low aromatic amino acid content. The protein subunit composition of the vesicles was determined by gel electrophoresis in (i) 0.1% sodium dodecyl sulfate at pH 9.0 and (ii) 5 M urea at pH 2.0. The membrane appeared to consist of one protein subunit of MW 50 000 daltons. Charge isomers of this subunit were not detected on urea gels. Antiserum prepared against purified gas vesicles of M. aquaticus S1 cross-reacted with the gas vesicles of all other gas vacuolate strains of M. aquaticus, as well as those of Prosthecomicrobium pneumaticum, Nostoc muscorum, and Anabaena flos-aquae, indicating that the gas vesicles of these widely divergent organisms have some antigenic determinants in common.Abbreviations SDS sodium dodecyl sulfate - MW molecular weight - Tris tris(hydroxymethyl)aminomethane - EDTA disodium ethylenediaminetetraacetic acid - BSA bovine serum albumin - TCA trichloroacetic acid - P _c pressure necessary to collapse gas vesicles     

Stereoselective formation of a K-region dihydrodiol from phenanthrene by Streptomyces flavovirens

The metabolism of phenanthrene, a polycyclic aromatic hydrocarbon (PAH), by Streptomyces flavovirens was investigated. When grown for 72 h in tryptone yeast extract broth saturated with phenanthrene, the actinomycete oxidized 21.3% of the hydrocarbon at the K-region to form trans-9,10-dihydroxy-9,10-dihydrophenanthrene (phenanthrene trans-9,10-dihydrodiol). A trace of 9-phenanthrol was also detected. Metabolites isolated by thin-layer and high performance liquid chromatography were identified by comparing chromatographic, mass spectral, and nuclear magnetic resonance properties with those of authentic compounds. Experiments using [9-¹⁴C]phenanthrene showed that the trans-9,10-dihydrodiol had 62.8% of the radioactivity found in the metabolites. Circular dichroism spectra of the phenanthrene trans-9,10-dihydrodiol indicated that the absolute configuration of the predominant enantiomer was (–)-9S,10S, the same as that of the principal enantiomer produced by mammalian enzymes. Incubation of S. flavovirens with phenanthrene is an atmosphere of ¹⁸O₂, followed by gas chromatographic/mass spectral analysis of the metabolites, indicated that one atom from molecular oxygen was incorporated into each molecule of the phenanthrene trans-9,10-dihydrodiol. Cytochrome P-450 was detected in 105,000×g supernatants prepared from cell extracts of S. flavovirens. The results show that the oxidation of phenanthrene by S. flavovirens was both regio- and stereospecific.Abbreviations CD circular dichroism - DMF N,N-dimethyl-formamide - GC/MS gas chromatography/mass spectrometry - HPLC high performance liquid chromatography - NMR nuclear magnetic resonance - ODS octadecylsilane - PAH polycyclic aromatic hydrocarbon - TLC thin-layer chromatography - TMS tetramethylsilane - UV ultraviolet     

A study of Salvia fruticosa Mill subsp. thomasii (Lacaita) Brullo,Guglielmo, Pavone & Terrasi,an endemic Sage of Southern Italy

The aim of this study was to investigate the essential oil profile of Salvia thomasii (Southern Italy) and to assess the relationship of this taxonomic unit to S. fruticosa. The essential oil, obtained by hydrodistillation, was analysed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The main components are β-pinene, viridiflorol, 1,8-cineole, camphor, α-pinene and trans-caryophyllene. The oil was tested for its potential antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Ability Power (FRAP) and β-carotene bleaching test. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity assays were used to screen the neuroprotective effect of the essential oil. A clear distinction of the two taxonomic units emerge by the comparison of S. thomasii and S. fruticosa s.l. essential oil composition. Considering the weak morphological differences and the clear-cut geographic separation we propose to treat the Italian unit at subspecies level as Salvia fruticosa Mill subsp. thomasii (Lacaita) Brullo, Guglielmo, Pavone & Terrasi.     

Epidermal structures and composition of epicuticular waxes of Sedum album sensu lato (Crassulaceae) in Balkan Peninsula

Sedum album is a succulent plant, with phenotypic variability resulting in delimitation of a number of taxa with diverse taxonomic recognition. We analyzed the taxa from Sedum album in broad sense (S. album, S. micranthum, S. athoum and S. serpentini) from Balkan Peninsula, to check whether the variability of their epidermal structures and epicuticular wax composition corresponds to their taxonomic reliability. Epidermal structures and epicuticular waxes were analyzed using light and scanning electron microscopy, gas chromatography-mass spectrometry and gas chromatography-flame ionization detector chromatography and multivariate statistical techniques. Analyses of studied taxa have shown that significant systematic characters include adaxial epidermal and guard cell length, stomatal frequency and subsidiary cells area on abaxial epidermis. The content of n-alkanes C₂₉, C₃₀, C₂₇, C₃₂ and C₃₃ was shown to be valuable criterion for their phytochemical delimitation. The comparison of epicuticular wax microstructures has shown no significant differences. Results indicate presence of two (micromorphology) or three (phytochemistry) well-defined groups of populations, with S. serpentini manifesting a considerable level of separation toward the others. Sedum athoum have shown weak to moderate degree of differentiation, while S. micranthum has shown general resemblance to S. album.     

Microbial conversion of tamoxifen

Summary Screening studies of tamoxifen (TAM) have shown that Streptomyces rimosus ATCC 2234 was able to metabolize TAM to 4-hydroxytamoxifen, which was obtained in a 10% yield and identified by two-dimensional nuclear magnetic resonance (NMR) spectroscopy. Several other microorganisms were capable of biotransforming TAM to tamoxifen-N-oxide and desmethyltamoxifen. The isolated metabolites were identified by gas chromatography/mass spectrometry and by two-dimensional NMR spectroscopic techniques. This is the first report of microbial hydroxylation of TAM. Offprint requests to: S. H. El-Sharkawy     

Mycotoxin formation by different geographic isolates of Fusarium crookwellense

Eighteen Fusarium crookwellense isolates from the continents of Australia, Europe, and North America were compared for their ability to produce mycotoxins on corn at 25 °C after 2 weeks. Extracts from corn fermented with each Fusarium isolate were analyzed by thin-layer chromatography (TLC) and gas chromatography/mass spectroscopy (GS/MS) for mycotoxins. Toxins detected were zearalenone (13 isolates), fusarin C (11 isolates), nivalenol (4 isolates), and diacetoxyscirpenol (2 isolates). Zearalenone and fusarin C were produced by isolates from each continent, while nivalenol was detected in the Fusarium isolates originating from Australia and one isolate from the United States.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned     

Metabolism of O-Ethyl S,S-Diphenyl Phosphorodithioate (Hinosan®) by Mycelial Cells of Pyricularia oryzae

Metabolism of organophosphorus fungicide Hinosan^® (O-ethyl S, S-diphenyl phosphorodithioate) by mycelia of P. oryzae, rice blast fungus, was studied using ³²P–, ³⁵S– and non-labeled compounds, by ion exchange chromatography, paper chromatography, thin-layer chromatography and gas chromatography, and identifying the metabolites and their derivatives with authentic compounds.

The main metabolic pathway is hydrolysis of one P-S linkage followed by the other P-S linkage or ethyl ester linkage and finally yielding phosphoric acid. A part of the fungicide metabolizes to hydroxylated intermediate metabolite, O-ethyl S-p-hydroxyphenyl S-phenyl phosphorodithioate. No significant difference in rate and mode of metabolism was found in this experiment between susceptible and resistant clones against the fungicide.     

(S)-雌马酚对肠道菌群的体外调控作用及其可代谢性研究

[目的]研究(S)-雌马酚对人体肠道菌群的体外调控作用和人体肠道菌群对(S)-雌马酚的代谢衍生作用。[方法]采用人体肠道菌群体外批量发酵、细菌16S rRNA基因高通量测序、气相色谱、液相色谱和质谱等检测(S)-雌马酚与人体肠道菌群体外相互作用。[结果]体外添加(S)-雌马酚对总体人肠道菌群结构和短链脂肪酸产量影响不明显。添加0.45 mmol/L (S)-雌马酚组与对照组相比,未检测到相对丰度发生显著变化的细菌;添加0.90 mmol/L (S)-雌马酚组与对照组相比,显著增加了肠杆菌科(Enterobacteriaceae)等条件致病菌的相对丰度,减少了潜在益生菌粪球菌属(Coprococcus)的比例。代谢分析发现,发酵培养液中(S)-雌马酚的浓度降低了约15%−30%,推测可能被微生物进一步降解或衍生修饰。[结论]从体外调控肠道菌群的角度判断,0.45 mmol/L (S)-雌马酚相对较安全,而0.90 mmol/L (S)-雌马酚可能会破坏肠道菌群平衡。(S)-雌马酚可以被人体肠道菌群进一步代谢,其特定代谢产物的结构与功能及其体内生物安全性有待进一步研究。     

Antibacterial Potential of 2,4-Di-tert-Butylphenol and Calixarene-Based Prodrugs from Thermophilic Bacillus licheniformis Isolated in Algerian Hot Spring

The present investigation reports the isolation, molecular identification and structure elucidation of antibacterial produced by two thermophilic spore-forming bacteria from hot spring (98?°C) of Guelma (Algeria). Morphological, biochemical and physiological characteristics were carried out. The molecular identification by 16S rRNA and 16-23S rRNA ITS-PCR sequencing identified the thermophilic strains as Bacillus licheniformis with 99% of similarity with GenBank accession numbers KX100031 and KX100032. Phenotypic characterization has mentioned several differences between thermophilic isolates and Bacillus licheniformis ATCC 14580. The ability of the thermophilic spore- forming bacteria to produce antibacterial compounds against two multidrug resistance bacteria Pseudomonas aeruginosa (NR_0754828.1) and Staphylococcus aureus (NR_075000.1) in pure and mixed culture was investigated by Radial Diffusion Assay at 55?°C. Structural elucidation of actives compounds was carried out using gas chromatography–mass spectrometry analyses. Antibacterial potency of the thermophilic isolates might be due to the association between two phenolic compounds: 2,4-Di-tert-butyl-phenol as principal active compound and p-tert-butylcalix[4]arene as prodrugs comparing between gas chromatography–mass spectrometry analysis of pure and mixed extract. To the best of our knowledge, this is the first report showing production of p-tert-butylcalix[4]arene and 2,4-Di-tert-butyl-phenol as extremolytes compounds from thermophilic Bacillus licheniformis at 55?°C.     

Identification of ononitol and O-methyl-scyllo-inositol in pea root nodules

Ononitol (4-O-methyl-myo-inositol) and O-methyl-scyllo-inositol were identified in pea (Pisum sativum L.) root nodules formed by twoRhizobium leguminosarum strains. Ononitol was the major soluble carbohydrate in nodules formed by strain 1045 while O-methyl-scyllo-inositol and two unidentified components were dominant in the carbohydrate pattern of the nodules formed by strain 1 a. The cyclitols were also present in the denodulated roots, but to a much smaller extent; in the above-ground plant parts only traces were found. The identification of ononitol and O-methyl-scyllo-inositol was established by gas chromatography and gas chromatography-mass spectrometry utilizing trimethylsilyl- and acetyl-derivatives.Abbreviations GC-MS gas chromatography-mass spectroscopy - TLC thin-layer chromatography     

Separation and Qualitative Study of Mentha asiatica Boriss. Essential Oil Components by prep-GC,GC-QTOF-MS,NMR and Prediction of Bioactive Compounds

In order to understand the material basis of wild Mentha asiatica Boris. in Xinjiang, the chemical constituents of essential oil extracted from aerial parts of this plant were studied. A total 52 components were detected and 45 compounds were identified. First of all, the essential oil was separated by silica gel column chromatography, and divided into several parts according to the results of thin layer chromatography. Eight fractions were obtained, and then each fragment was preliminarily screened for antibacterial activity. It was found that all eight fragments had certain antibacterial activity in different level. Then the fractions were subjected to preparative gas chromatography (prep-GC) for further isolation. Ten compounds were identified by ¹³C-NMR, ¹H-NMR and gas chromatography-quadrupole time of flight-Mass spectrometry (GC-QTOF-MS). They are sabinene, limonene and β-caryophyllene, (1R*,3S*,5R*)-sabinyl acetate, piperitone oxide, rotundifolone, thymol, piperitone, 4-hydroxypiperiditone, cedrol. After screened by bioautography, 4-hydroxypiperone and thymol were showed best antibacterial activity. The inhibitory effects of the two isolated compounds on Candida albicans and their related mechanisms were studied. The results showed that, 4-hydroxypiperone and thymol significantly reduced ergosterol content on the surface of Candida albicans cell membrane in a dose-dependent manner. This work has accumulated experience for the development and utilization of Xinjiang characteristic medicinal plant resources and new drug research and development, and provided scientific basis and support for the later research and development of Mentha asiatica Boris.     

Abscisic Acid in Prunus Trees: Isolation and the Effect on Growth of Excised Shoot Tissue

The abscisic acid content of the mature shoot tissue of vigorous Prunus avium, medium sized Prunus cerasus and dwarfing Prunus cerasus self (S) was estimated by spectropolarimetry and gas chromatography. There is no correlation between the levels of ABA found in the tissue and the dwarfing potential of the trees investigated. However, Prunus arium stem tissue shows the lowest sensitivity to ABA, whereas the explants of normal and dwarfing Prunus cerasus exhibit a stronger inhibition.     

Antifungal effects of volatile organic compounds from the endophytic fungus Cryptosporiopsis ericae Cc-HG-7 isolated from Coptis chinensis Franch

In this study, 136 strains of endophytic fungi were isolated from the Chinese traditional medicinal plant Coptis chinensis Franch. Of these, 129 strains were classified into 12 different genera according to morphological traits and internal transcribed spacer (ITS) gene sequence analyses. Their antifungal activities were assessed against the following fungi: Magnaporthe oryzae, Pythium graminicola, Cylindrocarpon destructans, Fusarium oxysporum, Cercospora zeae-maydis, Sclerotinia sclerotiorum, Setosphaeria turcica and Botrytis cinerea. Fourteen endophytic strains were active against at least one of the selected fungi. The most active strain Cc-HG-7 identified as Cryptosporiopsis ericae displayed inhibition rates of 81.42% and 72.00%, respectively, against S. sclerotiorum and S. turcica in dual culture technique. The volatile antifungal compounds were identified using headspace solid-phase microextraction, followed by gas chromatography mass spectrometry to investigate the potential biocontrol mechanisms of strain Cc-HG-7. The results suggested that the strain Cc-HG-7 could be a potential agent for the biological control of S. sclerotiorum and S. turcica.     

Micromorphological and chemical characterisation of Stachys recta L. subsp. serpentini (Fiori) Arrigoni in comparison to Stachys recta L. subsp. recta (Lamiaceae)

Stachys recta L. is a very polymorphous species in which numerous subspecies were recognised. S. recta L. subsp. serpentini (Fiori) Arrigoni is a typical endemism growing on serpentine soils in northern Apennines and particularly in Tuscany (Italy). In order to contribute to a better knowledge of this plant and to its differentiation with respect to S. recta L. subsp. recta, the micromorphological characters (non-glandular and glandular trichomes) and the essential oil composition of the two subspecies were investigated. Micromorphological characters were studied using scanning and transmission electron microscopy, while light microscopy was used for histochemical observations. Essential oil analysis was carried out by gas chromatography and mass spectrometry.In the two examined taxa, the morphology and distribution of glandular and non-glandular trichomes, and the different essential oil composition, may be considered distinctive characters at subspecies level. This is consistent with the taxonomic classification considering S. recta subsp. serpentini a subordinate taxon of S. recta.     

Salvia officinalis L. Essential Oils from Spain: Determination of Composition,Antioxidant Capacity,Antienzymatic, and Antimicrobial Bioactivities

Four essential oils (EO s) from Salvia officinalis L. cultivated in Spain (Murcia Province) were analyzed by gas chromatography coupled with mass spectrometry (GC /MS ) to determine their relative and absolute compositions. The main components were α ‐thujone (22.8 – 41.7%), camphor (10.7 – 19.8%), 1,8‐cineole (4.7 – 15.6%), and β ‐thujone (6.1 – 15.6%). Enantioselective gas chromatography identified (?)‐α ‐thujone and (+)‐camphor as the main enantiomers in all the analyzed EO s. Furthermore, when the EO s were tested to determine their antioxidant activity against free radicals and as ferric reducing and ferrous chelating agents, all were seen to have moderate activity due to the compounds they contained, such as linalool or terpinene. Because of their known relation with inflammatory illnesses and Alzheimer 's disease, respectively, the inhibition of lipoxygenase and acetylcholinesterase was studied using the EO s. Some individual compounds also inhibited these enzymes. In addition, the studied EO s were able to inhibit the growth of Escherichia coli , Staphylococcus aureus , and Candida albicans . The characterization carried out increases our awareness of the possible uses of S . officinalis EO as natural additives in food, cosmetics, and pharmaceuticals.     

Bactericidal and antioxidant effects of essential oils from Satureja montana L., Myristica fragrans H. and Cymbopogon flexuosus

The extraction and characterization of the essential oils (EO) from Satureja montana L., Myristica fragrans H. and Cymbopogon flexuosus and the determination of their antibacterial and antioxidant activities were achieved. The EO were identified by gas chromatography/mass spectrometry and quantified by gas chromatography using a flame ionization detector. The antibacterial potential against Escherichia coli and Staphylococcus aureus was evaluated by cell susceptibility assays and by scanning electron microscopy. The antioxidant activity was evaluated by the 2,2-diphenyl-1-picrylhydrazyl assay, by β-carotene bleaching and by determining the reducing power. Borneol (36·18%), γ-terpineol (12·66%) and carvacrol (11·07%) were the principal components in the EO from S. montana, and sabinene (49·23%) and α-pinene (13·81%) were found in the EO from M. fragrans. Geranial (59·66%) and neral (38·98%) isomers were the only major components in the EO from C. flexuosus. The EO from S. montana was effective against E. coli, with minimum inhibitory and bactericidal concentrations (MIC and MBC) of 6·25 µl ml⁻¹, whereas bactericidal potential against both was observed for the EO from M. fragrans; MIC = 6·25 µl ml⁻¹ for S. aureus and MBC = 12·5 µl ml⁻¹ for E. coli. A significant protective role on lipid substrates in the β-carotene bleaching assay was seen for the EO from S. montana and M. fragrans. Overall, such EO can be promising agents against pathogenic bacteria and for protecting biomolecules during oxidative stress.