bluuebunny/crossref_metadata_2025 huggingface dataset:
Essential Oil of Daucus glaber Forssk
Authors: Mansour El-Sayed S. , Maatooq Galal T. , Khalil Ashraf T. , Marwan El-Sayed M. , Sallam Amal A.
Published: February 2015

The composition of the essential oil of the fruits, leaves and stems of Daucus glaber Forssk has been studied by GC/MS. It was found that, the essential oil of the fruits consists of monoterpene hydrocarbons (limonene and sylvestrene are the majors) and phenylpropanoids (elemicin is the major). Sylvestrene has never been reported before in the essential oil of any Daucus species. The study of the essential oil of the leaves revealed the presence of monoterpene hydrocarbons; limonene and γ-terpinene are the majors and a small amount of sylvestrene. The essential oil of stems consists of monoterpene hydrocarbons (γ-terpinene is the major), terpene alcohols (mainly 4-terpineol) and phenylpropanoids (myristicin and elemicin are the majors). It is interesting that, the essential oil of the fruits is free from any oxygenated terpenes while that of the stems is free from limonene and sylvestrene which are present in the essential oil of the fruits and leaves in fairly large amounts. The essential oil of the fruits, leaves and stems shows broad antimicrobial activities against both gram positive and gram negative bacteria. In addition, the volatile oil of the stem, particularly, show activi­ties against Candida albicans (yeast). Also, the prepared oils have variable cytotoxic activities with LC50 21.52, 36.01 and 42.34 μrg/ml. respectively.
Complete Reduction of 2H-Pyran-2-one Moiety of Coumarin and 6-Methyl Coumarin by Colletotrichum capsici
Authors: Krishna Kumari Gadepalli Narasimha , Krishna Kumari Gadepalli Narasimha , Ganesh Munuswamy Ramanujam , Anitha Rajasekar , Aravind Sivasubramanian
Published: February 2015

The microbial transformation of coumarin (1) and 6-methyl coumarin (2) using Colletotrichum capsici gave 2-(3′-hydroxypropyl) phenol (3) and 2-(3′-hydroxypropyl)-4-methyl phe­nol (4). The phytopathogenic fungi effectively reduced the 2H-pyran-2-one moiety of both parent coumarins to respective alcohols.
Modulation of Sodium Pumps by Steroidal Saponins
Authors: de Souza Aloa Machado , Silva Lara Lucienne da , Previato Jose Osvaldo , Lopes Aníbal Gil , Caruso-Neves Celso , da Silva Bernadete Pereira , Paz Parente José
Published: February 2015

Costus spicatus, used in Brazilian traditional medicine to expel kidney stones, contains steroidal saponins with different chemical characteristics. In spite of its popular utilization as potent diuretic, no scientific reports correlate this activity with the chemical constituents of the extract. Therefore, two steroidal saponins (3β,22a,25R)-26-(β-ᴅ-glucopyranosyloxy)-2-methoxyfurost-5-en-3-yl O-ᴅ-apio-β-ᴅ-furanosyl-(1→2)-O-[6-deoxy-α-ʟ-mannopyranosyl- (1→4)]-β-ᴅ-glucopyranoside (1) and (3β,22α,25R)-spirostan-3-yl O-ᴅ-apio-β-ᴅ-furanosyl- (1→2)-O-[6-deoxy-α-ʟ-mannopyranosyl-(1→4)]-β-ᴅ-glucopyranoside (la), were isolated from the rhizomes of this plant and their effects on the Na+-ATPase and (Na++K+)-ATPase activities of the proximal tubule from pig kidney were evaluated. It was observed that 1 and la inhibit specifically the Na+-ATPase activity.
Comparison of the Volatile Composition of Stachys persica Gmel. and Stachys byzantina C. Koch. Oils Obtained by Hydrodistillation and Steam Distillation
Authors: Khanavi Mahnaz , Hadjiakhoondi Abbas , Amin Gholamreza , Amanzadeh Yaghoob , Rustaiyan Abdolhossein , Shafiee Abbas
Published: February 2015

The oils obtained by hydrodistillation and steam distillation of the aerial parts of Stachys persica Gmel. and Stachys byzantina C. Koch grown in Iran were analyzed by GC/MS. The essential oil obtained by hydrodistillation of the aerial parts of S. persica was characterized by a high amount of non-terpenoid components of which methyllinoleate (27.7%), hexadecanoic acid (9.8%) and 6,10,14-trimethyl-2-pentadecanone (9.2%) were the major constituents, whereas the steam distilled oil of the plant contained hexadecanoic acid (27.2%), carvacrol (9.4%) and eugenol (5.2%).

Both hydrodistilled and steam distilled essential oils of the aerial parts of S. byzantina were rich in sesquiterpenes such as α-copaene (16.6% and 10.4%), spathulenol (16.1% and 18.5%) and β-caryophyllene (14.3% and 13.5%), respectively.
Antioxidant and Immunomodulatory Constituents of Henna Leaves
Authors: Mikhaeil Botros R. , Badria Farid A. , Maatooq Galal T. , Amer Mohamed M. A.
Published: February 2015

The immunomodulatory bioassay-guided fractionation of the methanolic extract of henna (Lawsonia inermis L.; syn. Lawsonia alba L.) leaves resulted in the isolation of seven compounds; three have been isolated for the first time from the genus, namely p-coumaric acid, 2-methoxy-3-methyl-1,4-naphthoquinone and apiin, along with the previously isolated compounds: lawsone, apigenin, luteolin, and cosmosiin. Structural elucidation of the isolated compounds was based upon their physical, chemical as well as spectroscopic characters. Their immuomodulatory profile was studied using an in vitro immunoassay, the lymphocyte transformation assay. The ABTS [2,2′-azino-bis (3-ethyl benzthiazoline-6-sulfonic acid)], free radical scavenging assay depicted that all isolated compounds exhibited antioxidant activity comparable to that of ascorbic acid.
Bioactivity of Lignans from Taxus baccata
Authors: Erdemoglu Nurgun , Sener Bilge , Choudhary M. Iqbal
Published: February 2015

Three lignan derivatives, (-)-taxiresinol (1), (-)-3′-demethylisolariciresinol-9′-hydroxyisopropylether (2) and (-)-3-demethylisolariciresinol (3), previously isolated from the heartwood of Taxus baccata L. (Taxaceae) were investigated for cytotoxicity against the Oncology Cell Line Panel (breast, colon, ovary, prostate, lung and a normal adult bovine aortic endothelial cell line) as well as for antimicrobial activities. Besides, a chloroform-soluble portion of the ethanol extract from the heartwood of T. baccata was also tested for antimicrobial activities. Compounds 1 - 3 did not demonstrate much cytotoxic potency according to the reference drug etoposide. With the exception of compound 2, the other two lignans (1 and 3) and the chloroform extract were shown to possess antifungal activity, whereas only the chloroform extract exhibited antibacterial activity against Pseudomonas aeruginosa
Binary Mixtures of (N-phosphonomethyl)-glycine with New Aminophosphonates
Authors: Bielecki Krzysztof , Sarapuk Janusz , Kleszczynáska Halina
Published: February 2015

The potential biological activity of binary mixtures of some new organophosphorous compounds, aminoalkane- and aminofluorenephosphonates, with (N-phosphonomethyl)-glycine (glyphosate, PMG) was studied. The inhibition of growth of wheat (Triticum aestivum) induced by individual compounds and their equimolar mixtures with PMG was a measure of that activity. The experiments were expected to show if the new compounds exhibited good biological activity to be used for agrochemical applications and if this activity can be improved when they are used in mixtures with glyphosate which is the active component of the well-known herbicide Roundup.

The results obtained show that aminofluorenephosphonates inhibited wheat growth when used in micromolar concentrations. Thus, their efficiency can be compared to that of PMG. The efficiency of aminoalkanephosphonates was one order of magnitude weaker. The measure of the efficiency was the effective concentration inhibiting wheat growth by 50% (EC50). The most demanded interaction, i.e., a synergistic was observed for only one of binary mixtures of the compounds studied with PMG. Mostly they showed antagonistic or strong antagonistic interactions. Some of them were of the additive type. Such results exclude the possibility of potential use of all the compounds studied in binary mixtures with phosphonomethylglycine, especially as the mentioned synergistic interaction found was rather weak. The influence of structural features of aminophosphonates on the results obtained is discussed.
Green Paramecia as an Evolutionary Winner of Oxidative Symbiosis: A Hypothesis and Supportive Data
Authors: Kawano Tomonori , Kadono Takashi , Kosaka Toshikazu , Hosoya Hiroshi
Published: February 2015

A single cell of the green paramecia (Paramecium bursaria) harbors several hundreds of endo-symbiotic Chlorella-like algae in its cytoplasm. Removal of algae from the host organism and re-association of ex-symbiotic host paramecia with ex-symbiotic algae can be experimentally demonstrated in the laboratory. However, the mechanism precisely governing the alga-protozoan association is not fully understood, and the origin of symbiosis in the evolutionary view has not been given. Here, we propose the possible biochemical models (models 1 and 2) explaining the co-evolution between Paramecium species and algal symbionts by pointing out that algal photosynthesis in the host paramecia plays a dual role providing the energy source and the risk of oxidative damage to the host. Model 1 lays stress on the correlation between the (re)greening ability of the paramecia and the tolerance to oxidative stress whereas model 2 emphasizes the cause of evolutionary selection leading to the emergence of Paramecium species tolerant against reactive oxygen species.
Rosmarinic Acid Synthesis in Transformed Callus Culture of Coleus blumei Benth
Authors: Bauer Nataša , Leljak-Levanic Dunja , Jelaska Sibila
Published: February 2015

Agrobacteria mediated Coleus blumei tumour tissues were cultured in vitro on MS medium. Sixteen diversified transformed callus cultures were maintained for several years in the absence of plant growth regulators and antibiotics without affecting the growth rate. Rosmarinic acid was detected spectrophotometrically in all tissue lines but in different quantities. The highest rosmarinic acid accumulation detected was 11% of dry tissue mass. The relation between culture growth and rosmarinic acid production was investigated in three callus lines. The lines showed different rosmarinic acid accumulation in relation to their growth rate; it was either parallel or inversely related to the tissue growth. The effects of certain medium constituents on the callus growth and rosmarinic acid accumulation were examined in four tumour cell lines. Addition of 4% or 5% sucrose stimulated rosmarinic acid synthesis and decreased callus growth. Nitrogen reduction to one half or one quarter of initial concentration did not affect rosmarinic acid synthesis and decreased callus growth in three lines, while it increased rosmarinic acid accumulation and callus growth in one line. Addition of 0.1 mg/l Phe stimulated rosmarinic acid production in two lines but had little effect on the rosmarinic acid level in others. Rosmarinic acid production was significantly improved on modified macronutrients, where the Ac2 line produced 16.5 mg of rosmarinic acid per tube (0.2 g of dry wt) after being in culture for 35 days.
Cloning and Sequencing of Hydroxylase Genes Involved in Taxol Biosynthesis
Authors: Tu Jun , Zhu Ping , Cheng Ke-di , Meng Chao
Published: February 2015

Two full-length cDNAs (TCH1 and TCH2) were obtained from a cDNA library of Taxus chinensis mainly by the single specific-primer PCR (SSP-PCR) method. Compared with other reported enzymes from Taxus species, the deduced amino acid sequences of TCH1 and TCH2 exhibit significant homologies to hydroxylases that are involved in taxol biosynthesis. These findings imply that the two new genes are closely related to the biosynthesis of taxol/taxoids.
A Serine Protease from a Detergent-soluble Extract of Leishmania (Leishmania) amazonensis
Authors: Silva Lopez Raquel Elisa da , De Simone Salvatore Giovanni
Published: February 2015

Proteases mediate important crucial functions in parasitic diseases, and their characterization contributes to the understanding of host-parasite interaction. A serine protease was purified about 43-fold with a total recovery of 60% from a detergent-soluble extract of promastigotes of Leishmania amazonensis. The purification procedures included aprotinin-agarose affinity chromatography and gel filtration high performance liquid chromatography. The molecular mass of active enzyme was 110 kDa by native gel filtration HPLC and by SDSPAGE gelatin under non-reducing conditions. Under conditions of reduction using SDSPAGE gelatin analyses the activity of enzyme was observed in two proteins of 60 and 45 kDa, suggesting that the enzyme may be considered as a dimer. The Leishmania protease was not glycosylated, and its isoelectric point (pI) was around 4.8. The maximal protease activity was at pH 7.0 and 28 °C, using α-N-ᵨ-tosyl-ʟ-arginyl-methyl ester (ʟ-TAME) as substrate. Assays of thermal stability indicated that this enzyme was totally denatured after pre-treatment at 42 °C for 12 min and preserved only 20% of its activity after pre-treatment at 37 °C for 24 h, in the absence of substrate. Hemoglobin, bovine serum albumin (BSA), ovalbumin and gelatin were hydrolyzed by Leishmania protease. Inhibition studies indicated that the enzyme belonged to a serine protease class because of a significant impediment by serine protease inhibitors such as benzamidine, aprotinin, and antipain. The activity of the present serine protease is negatively modulated by calcium and zinc and positively modulated by manganese ions. This is the first study that reports the purification of a protease from a detergent-soluble extract of Leishmania species.
Total, Free and Conjugated Sterolic Forms in Three Microalgae Used in Mariculture
Authors: Mohammady Nagwa Gamal
Published: February 2015

Total, free and conjugated forms (steryl esters, steryl glycosides and acyl steryl glycosides) of sterols from three microalgae that are extensively used in mariculture (Tetraselmis chuii, Nannochloropsis salina and Skeletonema costatum) were examined. The results revealed that cholesterol is the only common fraction detected in all investigated species and distributed in free and all conjugated forms. However, the total sterol content of T. chuii was about 325 μg/g dry wt, most of it was concentrated amongst 24-methylcholesta-5,24-diene-3β-ol and 24-methylcholest-5-en-3β-ol. On the other hand, the majority of the fractions were distributed in the free form. The total sterol content of N. salina was about 180 μg/g dry wt, cholesterol was the major fraction that was detected. Nevertheless, the dominant distribution forms were esterified. While in S. costatum, the total sterol content was 76 μg/g dry wt, approximately most fractions are quantitatively alike and dominated in the free form. Furthermore, our study shows clearly that most sterols are not distributed regularly within each form, a result that encouraged us to suggest a distribution of specific sterol fraction as a free or conjugated can be used as a serving tool in chemotaxonomic studies.
Separation of Flavonoids from Aleurites moluccana Leaves Using Chitosan Modified with Heptaldehyde
Authors: Morsch Michele , Girardi Leury G. J. , Cechinel-Filho Valdir , Meyre-Silva Christiane , Rodrigues Clóvis Antonio
Published: February 2015

Heptaldehyde-modified chitosan (heptyl-chitosan, CH-Hp) was investigated as adsorbent for chromatograhic separation of the flavonoids from A. moluccana. The amount of 2″-Orhamnosylswertisin isolated (30.0 mg) was approx. twice as high as swertisin (17.5 mg). The improved surface hydrophobicity effected by the heptyl groups promoted the separation of flavonoids. From the results obtained, CH-Hp seems to be more suitable for separation of glycosylated flavonoids than other flavonoids. Thus, modified chitosan described here can be used for hydrophobic interaction chromatography as sucessfully illustrated with flavonoids.
The Influence of Growth Regulators on Membrane Permeability in Cultures of Winter Wheat Cells
Authors: Filek Maria , Biesaga-Kościelniak Jolanta , Marcińsk Izabela , Machačková Ivana , Krekule Jan
Published: February 2015

The effect of plant growth substances (IAA, 2,4-D, zeatin, kinetin, zearalenone) were studied on membrane properties of the cells of embryogenic (E) and non-embryogenic (NE) calli derived from immature inflorescences (inf) or embryos (emb) of winter wheat. Calli initiated from inflorescences show higher permeability. The ion leakage from cells of E calli was higher than from cells of NE calli. Growth regulators were used in concentrations of 2-30 mg/l (about 10-140 μm). All tested growth substances increased ion leakage from NE emb cells, IAA, zeatin and kinetin being most effective. In NE inf cells the effect of growth substances was similar as in NE emb, but much weaker. In E cells of both types (inf and emb) growth substances decreased ion leakage. Changes in the leakage of potassium and calcium ions were similar to those in total ion leakage. The uptake of labelled auxins (IAA and 2,4-D) was higher in NE cells (especially in NE inf) than in E cells. The endogenous level of IAA was higher in E cells than in NE cells and in inf cells than in emb cells. The importance of auxin in determining permeability of cell membranes is discussed.
Effect of Photon Flux Density and Temperature on the Production of Halogenated Monoterpenes by Plocamium cartilagineum (Plocamiaceae, Rhodophyta)
Authors: Palma Rodrigo , Edding Mario , Rovirosa Juana , San-Martín Aurelio , Argandoña Victor H.
Published: February 2015

The effect of different photon flux densities (PFD) and temperatures on the relative growth rate (RGR) and the concentration of three halogenated monoterpenes in samples of Plocamium cartilagineum L.( Dixon), a marine alga (Rhodophyceae), were studied. The highest RGR (22.8 ± 0.04 d-1) was obtained at 15 °C and 41 μmol m-2 s-1 of PFD and the lowest (18.0 ± 0.2 d-1) was obtained at 18 °C and 120 μmol m-2 s-1. The different temperatures and light used in assays did not affect significantly the production of organic compounds. The production of mertensene and violacene was not affected significantly. However, compound 1 reached the highest concentration at 15 °C and 65 μmol m-2 s-1. The relationship between growth and production of monoterpenes of P. cartilagineum and the effect of temperature and the PFD were analyzed.
Microsomal Detoxification Enzymes in Yam Bean [Pachyrhizus erosus (L.) Urban]
Authors: Belford Ebenezer J. D. , Dörfler Ulrike , Stampf Andreas , Schröder Peter
Published: February 2015

Cytochrome P450s and glutathione-S-transferases (GSTs) constitute two of the largest groups of enzyme families that are responsible for detoxification of exogenous molecules in plants. Their activities differ from plant to plant with respect to metabolism and substrate specificity which is one of the reasons for herbicide selectivity. In the tuber forming yam bean, the legume Pachyrhizus erosus, their activities at the microsomal level were investigated to determine the detoxification status of the plant. The breakdown of the herbicide isoproturon (IPU) to two distinct metabolites, 1-OH-IPU and monodesmethyl-IPU, was demonstrated. GST activity was determined with model substrates, but also by the catalysed formation of the fluorescent glutathione bimane conjugate. This study demonstrates for the first time microsomal detoxification activity in Pachyrhizus and the fluorescence image description of microsomal GST catalysed reaction in a legume.
Interactions of Calf Spleen Purine Nucleoside Phosphorylase with 8-Azaguanine, and a Bisubstrate Analogue Inhibitor: Implications for the Reaction Mechanism
Authors: Wierzchowski Jacek , Bzowska Agnieszka , Stępniak Katarzyna , Shugar David
Published: February 2015

Interactions of calf spleen purine nucleoside phosphorylase (PNP) with a non-typical substrate, 8-azaguanine (8-azaG), and a bisubstrate analogue inhibitor, 9-(2-phosphonylmethoxyethyl)- 8-azaguanine (PME-azaG), were investigated by means of steady-state fluorescence spectroscopy. Both 8-azaG and PME-azaG form fluorescent complexes with the enzyme, and dissociation constants are comparable to the appropriate parameters (Km or Ki) obtained from kinetic measurements. PME-azaG inhibits both the phosphorolytic and synthetic pathway of the reaction in a competitive mode. The complex of 8-azaG with PNP is much weaker than the previously reported Gua-PNP complex, and its dissociation constant increases at pH > 7, where 8-azaG exists predominantly as the monoanion (pKa ≈ 6.5). The fluorescence difference spectrum of the PNP/8-azaG complex points to participation of the N(7)H or/and N(8)H tautomers of the neutral substrate, and the 9-(2-phosphonylmethoxyethyl) derivative also exists as a neutral species in the complex with PNP. The latter conclusion is based on spectral characteristics of the PNP/PME-azaG complex, confirmed by fluorimetric determination of dissociation constants, which are virtually pH-independent in the range 6-7. These findings testify to involvement of the neutral purine molecule, and not its monoanion, as the substrate in the reverse, synthetic reaction. It is proposed that, in the reverse reaction pathway, the natural purine substrate is bound to the enzyme as the neutral N(7)H tautomer, which is responsible for the reported strong fluorescence of the guanine-PNP complex.
6-Substituted Indanoyl Isoleucine Conjugate Induces Tobacco Plant Responses in Secondary Metabolites
Authors: Hu Qun , Boland Wilhelm , Liu Ji-Kai
Published: February 2015

To characterize the role of the phytotoxin mimic 6-substituted indanoyl isoleucine conjugate 1 in plant secondary metabolism, tobacco (Nicotiana tabacum L. K326) was treated with compound 1. The volatile compounds of tobacco leaves were analyzed by GC-MS. In contrast to the control, three compounds, farnesene (2), santalol (3) and tetradecanal (4), were induced by treatment with 1 mm of compound 1. Concurrently other volatile compounds were also regulated.
Bioactive Constituents from Dracocephalum subcapitatum (O. Kuntze) Lipsky
Authors: Saeidnia Soodabeh , Gohari Ahmad Reza , Ito Michiho , Kiuchi Fumiyuki , Honda Gisho
Published: February 2015

From an EtOAc extract of Dracocephalum subcapitatum, five flavonoids, calycopterin, xanthomicrol, isokaempferide, luteolin and apigenin, together with five terpenoids, oleanolic acid, ursolic acid, geranial, neral and limonene-10-al, were isolated. Among them, citral and limonene-10-al were the most effective components against epimastigotes of Trypanosoma cruzi, the parasitic agent of Chagas disease.
Antibacterial Activity and Chemical Composition of the Essential Oil of Grammosciadium platycarpum Boiss. from Iran
Authors: Sonboli Ali , Eftekhar Fereshteh , Yousefzadi Morteza , Kanani Mohammad Reza
Published: February 2015

The chemical composition of the essential oils obtained from two samples (GP1 and GP2) of Grammosciadium platycarpum Boiss. was analyzed by GC and GC-MS. The analysis of the oils resulted in the identification of twenty-two constituents. Linalool (79.0% - GP1, 81.8% - GP2) and limonene (10.0%, 5.8%) were found to be the major components, respectively. The in vitro antibacterial activities of these oils and their main compounds against seven Gram-positive and Gram-negative bacteria were investigated. The results exhibited that the total oils and their major components possess strong to moderate activities against all the tested bacteria except for Pseudomonas aeruginosa.
Topical Anti-inflammatory Activity of Flavonoids and a New Xanthone from Santolina insularis
Authors: Cottiglia Filippo , Casu Laura , Bonsignore Leonardo , Casu Mariano , Floris Costantino , Sosa Silvio , Altinier Gianmario , Loggia Roberto Della
Published: February 2015

Bioactivity-guided fractionation of the methanol extract from the leaves of Santolina insularis led to the isolation of one new xanthone, (E)-3-{6-[(E)-3-hydroxy-3-oxo-1-propenyl]-9- oxo-9H-xanthen-2-yl}-2-propenoic acid, together with six known flavonoids: hispidulin, nepetin, cirsimaritin, rhamnocitrin, luteolin and luteolin 7-O-β-ᴅ-glucopyranoside. The structures were elucidated by means of 1D-, 2D-NMR spectroscopy and mass spectrometry. The topical anti-inflammatory activity of all isolated compounds and extracts was investigated employing the croton oil-induced dermatitis in mouse ear. The most active compound, luteolin, showed an ID50 of 0.3 μmol/cm2 and prevented ear oedema more effectively than an equimolar dose of indomethacin within 24 h.
Effects of Kaurane Diterpene Derivatives on Germination and Growth of Lactuca sativa Seedlings
Authors: Vieira Henriete S. , Takahashi Jacqueline A. , Pimenta Lúcia P. S. , Boaventura Maria Amélia D.
Published: February 2015

Kaurenoic and grandiflorenic acid, isolated from Wedelia paludosa (Asteraceae), some derivatives from these acids (alcohols, esters, amides, lactones, oximes) and other naturally occurring kaurane diterpenes were tested for their action on the growth of radical and shoot of Lactuca sativa. Gibberellic acid, GA3, a commercially available phytohormone, belonging to the same class of diterpenes, was also tested. Some of the tested substances showed a remarkable activity either in the inhibition or in stimulation of L. sativa growth. The activity, in some cases, was even higher than that of GA3.
Morning Glory Systemically Accumulates Scopoletin and Scopolin after Interaction with Fusarium oxysporum
Authors: Shimizu Bun-ichi , Miyagawa Hisashi , Ueno Tamio , Sakata Kanzo , Watanabe Ken , Ogawa Kei
Published: February 2015

An isolate of non-pathogenic Fusarium, Fusarium oxysporum 101-2 (NPF), induces resistance in the cuttings of morning glory against Fusarium wilt caused by F. oxysporum f. sp. batatas O-17 (PF). The effect of NPF on phenylpropanoid metabolism in morning glory cuttings was studied. It was found that morning glory tissues responded to treatment with NPF bud-cell suspension (108 bud-cells/ml) with the activation of phenylalanine ammonialyase (PAL). PAL activity was induced faster and greater in the NPF-treated cuttings compared to cuttings of a distilled water control. High performance liquid chromatography analysis of the extract from tissues of morning glory cuttings after NPF treatment showed a quicker induction of scopoletin and scopolin synthesis than that seen in the control cuttings. PF also the induced synthesis of these compounds at 105 bud-cells/ml, but inhibited it at 108 budcells/ ml. Possibly PF produced constituent(s) that elicited the inhibitory effect on induction of the resistance reaction. These compounds could potentially be useful as markers to detect early beginning interactions between Fusarium and morning glory tissues cuttings.
Pigments and Citrinin Biosynthesis by Fungi Belonging to Genus Monascus
Authors: Pisareva Emiliya , Savov Valentin , Kujumdzieva Anna
Published: February 2015

Citrinin is a mycotoxin, which is produced by fungi belonging to the genus Monascus, known in biotechnology as producers of azaphilone pigments. The relation between biosynthesis of these secondary metabolites was investigated in different species of the genus Monascus in batch-culture at the following cultivation conditions: T = 28 °C, agitation 220 rpm, and a medium, which induce citrinin production, containing ethanol as a carbon source.

The screening was carried out with 16 fungal strains and the biosynthesis of citrinin and pigments was monitored quantitatively at the standard conditions mentioned above. Some kinetic parameters of the process have been determined. The values of the growth yield coefficient YX/C were between 0.32 and 0.57. The amount of the extracellular red and orange pigments at the end of cultivation varied for the different strains between 0.09 and 1.33 OU/ mg dry weight, and 0.15 and 0.96 OU/mg dry weight, respectively. The amount of the total pigments measured was between 0.16 and 3.6 OU/mg dry weight, and between 0.21 and 3.39 OU/mg dry weight. The determined ratio 500 nm/400 nm, characterizing the pigment production, ranged between 0.60 and 1.06. Twelve of the investigated strains produced citrinin and pigments, two of them produced only pigments. Two strains were not able to produce neither pigments nor citrinin.

Thus, the biosynthesis of citrinin appeared to be strain-specific and does not correlate with the pigments’ biosynthesis by the fungal strains belonging to the genus Monascus.
Metabolism of 4-n-Nonylphenol by Non-modified and CYP1A1- and CYP1A2-Transgenic Cell Cultures of Tobacco
Authors: Berger Angelika , Ruß Alexander S. , Schuphan Ingolf , Schmidt Burkhard
Published: February 2015

The metabolism of 14C-4-n-nonylphenol (14C-4-n-NP), as a mo-el for the xenoestrogen nonylphenol, was investigate- in three types of tobacco cell suspension cultures: one genetically non-modified culture (NT) and two cultures constitutively expressing human cytochrome P450 CYP1A1 or CYP1A2. With 1 mg l-1 of 14C-4-n-NP and 24 h of incubation, the xenobiotic was transformed almost completely to glycosides. After glycosidic cleavage, 14C-4-n-NP and several primary metabolites of 14C-4-n-NP were liberated. Portions of the primary metabolites were 29.3% (NT culture), 34.3% (CYP1A1 culture), and 50.7% of applied 14C(CYP1A2 culture). Thus, the endogenous capacity of the tobacco cells to form primary metabolites of 4-n-NP was noticeably higher than that of CYP1A1 or CYP1A2. The results however clearly suggest that 4-n-NP is - even though a poor - substrate of CYP1A1 an- CYP1A2. In order to examine metabolic profiles of 4-n-NP in the NT, CYP1A1 and CYP1A2 cultures, the suspensions were exposed to 10 mg l-1 of 14C-4-n-NP using a two-liquid-phase system with carrier n-hexadecane and 192 h of incubation. Results obtained resembled those of the low concentration study. The oxidative metabolic profiles determined after hydrolytic cleavage using GC-EIMS were similar in the NT, CYP1A1 and CYP1A2 cultures. Main metabolites were side-chain mono-hydroxylated derivatives of 4-n-NP with 6′-, 7′- and 8′- OH-4-n-NP as prominent metabolites. In addition, olefinic side-chain hydroxy, ring methoxylated, keto and ring hydroxylated derivatives were observed. The lack of differences in metabolic profiles among the CYP1A1, CYP1A2 and NT cultures was referred to the low enzymatic activity of CYP1A1 and CYP1A2 as compared to the higher endogenous oxidative capacity of tobacco, as well as to similar metabolic profiles of 4-n-NP produced by CYP1A1 and CYP1A2 an- tobacco itself.
New Host Plants of Erwinia amylovora in Bulgaria
Authors: Atanasova Iliana , Kabadjova Petia , Bogatzevska Nevena , Moncheva Penka
Published: February 2015

Nine strains of Erwinia amylovora were isolated from new host plants in Bulgaria D chokeberry and strawberry. The strains were characterized morphologically and biochemically using the API 20E and BIOLOG system. It was established that they showed three different API 20E metabolic profiles, not found by previous studies of E. amylovora. All strains were identified as E. amylovora due to their metabolic fingerprint patterns obtained by the BIOLOG system. The identification was confirmed by PCR amplification of a specific region of plasmid pEA29 and genome ams-region. This study is the first characterization and identification of E. amylovora strains isolated from chokeberry and strawberry by the API 20E and BIOLOG system and by polymerase chain reaction.
Age Related Differences in Lipophilic Compounds Found in Femoral Gland Secretions of Male Spiny-footed Lizards, Acanthodactylus erythrurus
Authors: López Pilar , Martín José
Published: February 2015

Although chemoreception plays an important role in social organization of many lizards, only a few studies have examined the chemicals found in secretions used for intraspecific communication. We report the composition of the secretion of the femoral glands of males of the spiny-footed lizard (Acanthodactylus erythrurus). On the basis of mass spectra, obtained by GC/MS, we identified 45 lipophilic compounds, including several alcohols ranging from 10 to 29 carbon atoms (mainly hexacosanol and tetracosanol), steroids (mainly cholesterol and dehydrocholesterol), n-C9 to n-C20 carboxylic acids, esters of carboxylic acids, and minor components such as lactones, ketones, squalene and α-tocopherol. Some of these compounds are reported for the first time in lizards. Adult and subadult males differed in the composition of secretions, with C9 to C15 carboxylic acids being more abundant in younger than in older lizards, whereas C16 to C20 carboxylic acids were more abundant in older lizards. Also, older lizards had significant lower proportions of cholesterol and campesterol but higher proportions of dehydrocholesterol.
A Study on Differences between Radiation-induced Micronuclei and Apoptosis of Lymphocytes in Breast Cancer Patients after Radiotherapy
Authors: Taghavi-Dehaghani Mahnaz , Mohammadi Shahla , Ziafazeli Tahereh , Sardari-Kermani Manouchehr
Published: February 2015

Cancer patients’ responses to radiotherapy vary in severity. It has been suggested that it may be due to differences in intrinsic cellular radiosensitivity. Prediction of tissue reactions to radiotherapy would permit tailoring of dosage to each patient. Towards this goal the micronucleus and apoptosis tests have been proposed as methods for measurement of chromosomal damage in peripheral blood lymphocytes. In this study, gamma-ray sensitivity of cultured lymphocytes of 26 breast cancer patients with early or late reactions was investigated. After irradiation with 4 Gy gamma radiation in G0, the frequency of micronuclei for patients with early reactions was significantly higher (P < 0.05) than for patients with late reactions. In the contrary the frequency of apoptosis for patients with early reactions was significantly lower (P < 0.05) than in the other group. It could be suggested that such a reduced amount of micronuclei in the late effects group is due to the presence of some residual DNA damages which are not completely repaired and lesions show increasing severity when the patients’ cells are irradiated again. These induced damages, probably are high enough to stimulate other endpoints like apoptosis instead of micronuclei.
Two New Methods Monitoring Kinetics of Hydrolysis of Acetylcholine and Acetylthiocholine
Authors: Štěpánková Šárka , Vránová Martina , Zdražilová Pavla , Komers Karel , Komersová Alena , Čeganb Alexander
Published: February 2015

Hydroxylamine and HPLC methods, measuring in vitro kinetics of enzymatic hydrolysis of acetylcholine or acetylthiocholine by cholinesterases, are described. The hydroxylamine method determines the dependence of substrate concentration vs. time, the HPLC method is able to measure simultaneously the time dependences of substrate and both primary products, choline or thiocholine, and acetic acid. Practical determinations are shown, comparison with known (above all Ellman’s and pH-stat) methods, advantages and disadvantages are discussed.
A Ligand Function of Glutathione S-Transferase
Authors: Lederer Barbara , Böger Peter
Published: February 2015

Glutathione S-transferases (GSTs) are ubiquitous enzymes and abundant in plants. They are intimately involved in plant metabolism and stress defense related to reactive oxygen species. Our project assigned particular reactions including novel ones to certain GST-isoforms. Transformed E. coli was used to express recombinant GST-isoforms from maize. An N-terminal His tag allowed their purification by affinity chromatography. Three GST-monomers had a molecular weight of 26, 27, 29 kDa, and aggregated to dimers when assayed for their enzymic properties. Four dimeric isoforms were used to study how they interact with tetrapyrroles (of the chlorophyll biosynthesis pathway). It was found that protoporphyrin IX (Proto IX), Mg-protoporphyrin and other tetrapyrroles are bound non-covalently (“liganded”) to GSTs but not conjugated with reduced glutathione. This binding is non-covalent, and results in inhibition of conjugation activity, the degree depends on type of the porphyrin and GST-isoform. I50-values between 1-10 μᴍ were measured for Proto IX, the inhibition by mesoporphyrin and Mg-protoporphyrin was 2- to 5-fold less. The ligand binding is noncompetitive for the substrate 1-chloro-2,4-dinitrobenzene and competitive for glutathione. The dimer GST 26/26 prevents the (non-enzymic) autoxidation of protoporphyrinogen to Proto IX, which produces phytotoxic reactive oxygen species in the light. GST 27/27 protects hemin against degradation. Protoporphyrinogen is formed in the plastid and then exported into the cytosol. Apparently binding by a suitable GST-isoform ensures that the highly autoxidizable protoporphyrinogen can safely reach the mitochondrium where it is processed to cytochrome.
Insecticidal Activity of the Essential Oil of Ligusticum mutellina Roots
Authors: Passreiter Claus M. , Akhtar Yasmin , Isman Murray B.
Published: February 2015

The essential oil obtained from roots of different collections of Ligusticum mutellina was tested against 3rd instar armyworms, Pseudaletia unipuncta (Lepidoptera: Noctuidae), for insecticidal activity. The main compounds were isolated and their structures were elucidated using 2D-NMR techniques. Our collections contained dillapiole, ligustilide and myristicin as major compounds. The previously reported sarisan was not present, moreover its occurrence in L. mutellina should be revised based on our findings.
β-Carboline and Quinoline Alkaloids in Root Cultures and Intact Plants of Peganum harmala
Authors: Zayed Rawia , Wink Michael
Published: February 2015

Alkaloid profiles of root and shoot cultures, seedlings and mature plants were analysed by capillary GLC and GLC-MS. β-Carboline alkaloids, such as harmine, harmaline dominate in normal and root cultures transformed by Agrobacterium rhizogenes, as well as in roots and fruits of the plant. In shoots, flowers and shoot cultures quinoline alkaloids such as peganine, deoxypeganine, vasicinone and deoxyvasicinone widely replace the β-carboline alkaloids. In root cultures, the formation of β-carboline alkaloids can be induced by methyljasmonate and several other elicitors indicating that these alkaloids are part of the reactive chemical defence system of Peganum harmala.
In vitro Micropropagation of Boswellia ovalifoliolata
Authors: Chandrasekhar Thummala , Hussain T. Mohammad , Jayanand Boddu
Published: February 2015

A protocol for micropropagation of Boswellia ovalifoliolata Bal & Henry (Burseraceae) was developed using cotyledonary nodal explant on Murashige and Skoog modified medium (MS). A comparative study of micropropagation with 6-benzyladenine, kinetin and thidiazuron along with 1-naphthalene acetic acid (0.054 μм) was conducted. The highest shoot multiplication (7.1 ± 0.2 shoots per node) was achieved in 50 d on MS supplemented with thidiazuron (2.72 μм). Excised shoot cuttings of 3.0 cm were placed on the MS basal medium supplemented with indole-3-acetic acid and indole-3-butyric acid alone and in combinations for rooting. Activated charcoal (100 mg l-1) and polyvinylpyrrolidone (40 mg l-1) were added to the medium to prevent browning of cultures. The regenerated plantlets have been successfully acclimatized and transferred to soil.
Antiproliferative Effects of Two Amides, Piperine and Piplartine, from Piper Species
Authors: Bezerra Daniel Pereira , Pessoa Cláudia , Moraes Manoel Odorico de , Silveira Edilberto Rocha , Lima Mary Anne Sousa , Martins Elmiro Francisca Juliana , Costa-Lotufo Letícia Veras
Published: February 2015

The present work evaluated the cytotoxicity of piplartine {5,6-dihydro-1-[1-oxo-3-(3,4,5- trimethoxyphenyl)-trans-2-propenyl]-2(1H)pyridinone} and piperine {1-[5-(1,3)-benzodioxol- 5-yl)-1-oxo-2,4-pentadienyl]piperidine}, components obtained from Piper species. The substances were tested for their cytotoxicity on the brine shrimp lethality assay, sea urchin eggs development, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay using tumor cell lines and lytic activity on mouse erythrocytes. Piperine showed higher toxicity in brine shrimp (DL50 = 2.8 d 0.3 μg/ml) than piplartine (DL50 = 32.3 d 3.4 μg/ml). Both piplartine and piperine inhibited the sea urchin eggs development during all phases examined, first and third cleavage and blastulae, but in this assay piplartine was more potent than piperine. In the MTT assay, piplartine was the most active with IC50 values in the range of 0.7 to 1.7 μg/ml. None of the tested substances induced hemolysis of mouse erythrocytes, suggesting that the cytotoxicity of piplartine and piperine was not related to membrane damage.
Transcriptomics and Proteomics: Tools for Optimising Phytoremediation Activities
Authors: Coleman Julian O. D. , Haslam Richard P. , Downie Andrew L.
Published: February 2015

Negligent industrial development has greatly contributed to environmental pollution through the contamination of water and soil by xenobiotic organic chemicals. Remedial strategies to deal with chemical pollution require reliable methods to identify and monitor contamination, as well as effective procedures to attenuate or to eliminate the pollutant. In the food chain, plants are ideally placed as early bio-indicators of environmental pollution as they experience and respond to environmental toxicants sooner than organisms at higher trophic levels. Furthermore, some plants are capable of detoxifying anthropogenic chemicals by metabolic transformation and could prove useful for the remediation of contaminated water and soil: so-called phytoremediation.

So far research technologies aimed at developing plants for bio-indication/bio-monitoring and for remediation have largely relied on standardised chemical and biochemical procedures to evaluate phytotoxicity, metabolic fate and persistence of organic pollutants in plants. The next stage in the evolution of these plant-based technologies is the improvement and optimisation of any innate phytoremediation activities identified in selected plants.

In general, uptake followed by metabolism and compartmentation is responsible for the detoxification of organic xenobiotics in plants. These are complex cellular systems that may be organised in well-defined pathways and are often controlled by large families of genes. In order to elucidate complex traits such as detoxification, an emerging idea is to make use of global approaches such as the new “omic” technologies to identify molecular changes in plant tissues exposed to specific organic xenobiotics. From expression profiles at the messenger RNA level, transcriptomics permit the identification of function-related gene clusters and at the protein level proteomics provide information on where, when and at what level specific proteins accumulate.

We conclude that these global approaches may be a useful way of widening screening capability to identify appropriate molecular markers that can be used to improve detoxification activity.
Recombinant p-Hydroxyphenylpyruvate Dioxygenase of High Activity
Authors: Lederer Barbara , Böger Peter
Published: February 2015

Inhibitors of p-hydroxyphenylpyruvate dioxygenase (HPPD) are bleaching compounds impairing the formation of colored carotenoids. This activity makes them promising candidates for herbicides. Detailed studies on enzyme-inhibitor complexes or on the binding niche of the enzyme have still to be performed. Enzyme preparation from plants is time-consuming and the yield is poor. This paper describes in relevant detail the preparation of recombinant enzyme from Arabidopsis thaliana with good yield and high specific activity.
Two-Stage System for Micropropagation of Several Genista Plants Producing Large Amounts of Phytoestrogens
Authors: Łuczkiewicz Maria , Piotrowski Arkadiusz
Published: February 2015

A two-stage method for in vitro propagation of six Genista species from shoot tips was developed. Multiple microshoot cultures were obtained by growing the shoot tip explants on Schenk and Hildebrandt medium supplemented with 9.84 μm 6-(γ,γ-dimethylallylamino)- purine and 0.99 μm thidiazuron. The best shoot elongation was achieved on Schenk and Hildebrandt medium containing 4.92 μm indole-3-butyric acid. The rooting of shoots brought best effects (100%) on Schenk and Hildebrandt medium with 2.68 μm 1-naphthaleneacetic acid. HPLC analysis indicated that six-month-old regenerated plants as well as the herb of intact plants produced a rich set of simple flavones (derivatives of luteolin and apigenin) and isoflavones (derivatives of genistein, daidzein, formononetin and biochanin A). Multiple microshoot cultures of all species produced no simple flavones at all. In vitro shoots accumulated selectively a rich group of phytoestrogens in the form of aglucones, glucosides and esters (derivatives of genistein and daidzein). Cultures obtained in vitro synthesized many times more isoflavones than the intact plants. In all shoots which were micropropagated the dominating compound was genistin (e.g. shoots of G. tinctoria D ca 3281.4 mg per 100 g dry weight). Possible influence of tissue differentiation on isoflavone content under in vitro and in vivo conditions is discussed.
Interaction between Aspterric Acid and Indole-3-acetic Acid on Reproductive Growth in Arabidopsis thaliana
Authors: Shimada Atsumi , Yamane Hisakazu , Kimura Yasuo
Published: February 2015

Application of indole-3-acetic acid (IAA) with a pollen growth inhibitor, aspterric acid (AA), results in the recovery of normal pollen development. In contrast, application of gibberellin (GA3) with AA do not induce normal pollen growth. In addition, application of different concentrations of IAA with AA shortens the period of growth from bolting to first flowering as compared to that treated with AA alone. Furthermore, stem length and number of flower bud treated with IAA and AA were similar to those of control. These results suggest, that IAA may play an important role in reproductive growth of A. thaliana.
Volatiles from Four Astragalus Species: Phenological Changes and their Chemotaxonomical Application
Authors: Platikanov Stefan , Nikolov Stefan , Pavlova Dolja , Evstatieva Ljuba , Popov Simeon
Published: February 2015

This paper shows the changes of the volatile compounds from four Astragalus species at three phenological stages: leaf development, flowering and fructification, which might be connected with the plant defense. After GC/MS analyses of Astragalus glycyphyllos L., A. hamosus L., A. cicer L. and A. spruneri Boiss., different groups of volatile compounds were found: hydrocarbons, alcohols, aldehydes and ketones, esters, terpenes, chlorinated compounds, etc. Identified volatiles were used for a cluster analysis in order to make chemotaxonomic conclusions for these evolutionary different species.
Sex Attractants for Six Moth Species of the Families Brachodidae, Choreutidae and Tineidae from Kazakhstan and Lithuania
Authors: Karalius Vidmantas , Mozūraitis Raimondas , Būda Vincas , Libikas Ilme , Borg-Karlson Anna-Karin
Published: February 2015

Sex attractants were established for one Brachodidae, three Choreutidae and two Tineidae moth species during field screening tests with (2E,13Z)-octadecadien-1-al, (2E,13Z)-, (3E,13Z)-, (3Z,13Z)-octadecadien-1-ols and their acetates (2E,13Z-18:Ald, 2E,13Z-, 3E,13Z-, 3Z,13Z-18:OH/OAc) as well as of binary mixtures of these compounds in West-Kazakhstan and Lithuania. Males of Brachodes appendiculata were attracted by 3E,13Z-18:OAc, Prochoreutis ultimana and P. myllerana by 2E,13Z-18:OH, Monopis palidella by 2E,13Z-18:Ald and Triaxomera fulvimitrella by binary mixtures of 3Z,13Z-18:OAc with either 3E,13Z-18:OH in the ratio of 5:5 or 3Z,13Z-18:OH in the ratio of 9:1 (v/v). The 3-component mixture composed of 2E,13Z-18:OH, 3Z,13Z-18:OH and 2E,13Z-18:Ald in the ratio 1:1:1 was developed to attract Prochoreutis sehestediana males. Attraction antagonists for B. appendiculata, P. ultimana and M. palidella were shown.
Spontaneous Transfection of Mammalian Cells with Plasmid DNA
Authors: Tsoncheva Vania L. , Todorova Kristina A. , Ivanov Ivan G. , Maximova Vera A.
Published: February 2015

A simple method for spontaneous transfection into mammalian cells (both adherent and suspension in culture) with plasmid DNA is described. This method does not require any specific DNA carrier or technical device and can be applied for obtaining both transient and stably transfected cells. The efficiency of spontaneous transfection is slightly lower in comparison with that of the conventional calcium phosphate and lipofectin transfection methods and does not depend on the type of cell culture used.