bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Fatty Acid Composition of Cell Homogenates and Isolated Chloroplasts in Acetabularia crenulata (Lamouroux)
Authors: Santiago-Vázquez Lory Z. , Jacobs Robert S.
Published: February 2015

Algal preparations from Acetabularia crenulata were analyzed for their fatty acid composition to establish the suitability of this alga as a model to study fatty acid oxidation and oxylipin biosynthesis. The work was based on two goals. The first goal of this study was to determine the contribution of fatty acids from contaminating bacteria and how this influenced the total fatty acid composition of cell homogenates of A. crenulata collected in the wild as compared to specimens cultured in sterile conditions. The major fatty acids detected for both specimens were palmitic (C16:0), palmitoleic (C16:1n-7), oleic (C18:1n-9), linoleic (C18:2n-6), linolenic (C18:3n-3), and octadecatetraenoic acid (C18:4n-3). Significant amounts of odd-chain fatty acids common to bacteria were not detected in either sample. Furthermore, branched-chain fatty acids, typical bacterial biomarkers, were not detected in either sample. Data suggest that bacteria do not greatly contribute to the total fatty acid pool of A. crenulata.

The second goal was to compare the fatty acid composition of cell homogenates with that of isolated chloroplasts. Comparatively speaking palmitoleic and octadecatetraenoic acid were found at significantly lower concentrations in the chloroplast whereas oleic and linolenic acid were found at significantly higher amounts in this organelle. Furthermore, the amount of hexadecatrienoic acid (C16:3), a fatty acid commonly esterified to monogalactosyldiacylglycerol (MGDG; lipid present at high concentrations inside the chloroplasts of algae), was present at very low concentrations in these plastids (0.7%). Typically green algal follow the “prokaryotic pathway” for MGDG biosynthesis where C18:3 is esterified at the sn-1 position of the glycerol backbone and C18:3 or C16:3 at the sn-2 position, making C16:3 a major fatty acid inside chloroplasts. Interestingly, our results suggest that chloroplasts of A. crenulata appear to follow the “eukaryotic pathway” for MGDG biosynthesis where C18:3 is both at the sn-1 and sn-2 position of MGDG. Taking into account the exceptions noted, the fatty acid composition for A. crenulata is similar to that reported for most chlorophytes.
Understanding Genetic Diversity of the Liver Fluke Fasciola hepatica
Authors: Dosay-Akbulut Mine , Trudgett Alan , Stanhope Michael
Published: February 2015

Economical breeding is important to obtain maximum gain from the breeding in the animal sector. The economic loss has to be eliminated or should be minimized. The liver fluke, Fasciola hepatica, present mostly in sheep and dairy cattle affect the yield of animals and even cause their death. To eliminate or minimize the impact of these parasites on the animals, it is important to understand the genetic diversity of the liver fluke populations and the relationship between parasite and host at regional bases. This research was carried out to determine diversity by sequence analysis of the mitochondrial ND1 gene and ribosomal ITS1 region.
Volatile Compounds in Shoulder Gland Secretions of Male Flying Foxes, Genus Pteropus (Pteropodidae, Chiroptera)
Authors: Wood William F. , Walsh Allyson , Seyjagat John , Weldon Paul J.
Published: February 2015

The shoulder gland secretions of captive males of the Indian flying fox (Pteropus giganteus), the little golden-mantled flying fox (P. pumilus), the island flying fox (P. hypomelanus), and the large flying fox (P. vampyrus) were examined by gas chromatography-mass spectrometry. Sixty-five compounds, including hydrocarbons, carboxylic acids, alcohols, aldehydes, ketones, esters, and amides, were identified among the four species. Many of these compounds, such as squalene, cholesterol, and C5-C16 straight- and branched-chain carboxylic acids, are typical of tetrapod epidermal products. Aldehydes, which were detected in all four Pteropus species, and some straight- and branched-chain ketones, which were detected in P. hypomelanus and P. pumilus, are known from other mammalian skin glands. Acetophenone, 4-acetoxyacetophenone, and 4-hydroxyacetophenone were observed in P. pumilus; the last compound comprised 37.1% of the total ion current. 2,3-Butanediol, a prominent component (5.2-19.3%) in the secretions of P. giganteus, P. hypomelanus, and P. pumilus, and C10 and C12 isopropyl esters and C10-C14 1-methylbutyl esters, observed in P. hypomelanus and P. vampyrus, have not previously been reported from vertebrates. α-Methyl-4-methoxybenzyl alcohol and dihydro-5-phenyl-2(3H)-furanone, from P. giganteus and P. pumilus, are new natural products. 1-Chloro-3-methyl-2-butene, another new natural product, and five C5 compounds exhibiting a similar isoprenoid structure were observed in P. giganteus. Striking contrasts were observed in the chemical profiles of the species we examined, with even general chemical classes differentially represented among them.
Seasonal Variation and Analgesic Properties of Different Parts from Curcuma zedoaria Roscoe (Zingiberaceae) Grown in Brazil
Authors: Pamplona Christiane Regina , Souza Marcia Maria de , Machado Marina da Silva , Filhoa Valdir Cechinel , Navarro Dionezine , Yunes Rosendo Augusto , Monache Franco Delle , Niero Rivaldo
Published: February 2015

This work describes the seasonal variation of curcumenol (1) and dihydrocurdione (2), two active terpenoids from different parts (roots, mother rhizome and rugous rhizome) of Curcuma zedoaria grown in Brazil. The analysis was carried out by high resolution gas chromatography, using external standards for determination. The results showed that both terpenoids are present in all the parts studied. However, C. zedoaria exhibited about three times more terpenoids in the mother rhizome in autumn than in other parts and seasons studied. The antinociceptive activity of the dichloromethane extracts from different parts and collected in different seasons was studied using the acetic acid-induced abdominal constriction model in mice. The extracts obtained from mother rhizome collected in autumn and winter at doses of 10 mg/kg body weight, i.p., caused considerable antinociceptive activity inhibiting 91.1 and 93.4% of the abdominal constrictions, respectively, whereas compounds 1 and 2 caused inhibitions of 64.0 and 46.0%, respectively. These results confirm that both compounds contribute to explain the antinociceptive effect of the plant but suggest that other compounds are also acting as analgesics
Anti-Inflammatory and Antinociceptive Activity of Flavonoids Isolated from Viscum album ssp. album
Authors: Orhan Didem Deliorman , Küpeli Esra , Yesilada Erdem , Ergun Fatma
Published: February 2015

Viscum album L. has been used in the indigenous systems of medicine for treatment of headache and some inflammatory diseases. In order to evaluate this information, antinociceptive and anti-inflammatory activities of the five flavonoids (5,7-dimethoxy naringenin or 4′,6′- dimethoxy chalcononaringenin) derivatives, isolated from the ethyl acetate fraction of the extract from V. album ssp. album, were investigated, namely 5,7-dimethoxy-flavanone-4′-O- β-ᴅ-glucopyranoside (1), 2′-hydroxy-4′,6′-dimethoxy-chalcone-4-O-β-ᴅ-glucopyranoside (2), 5,7-dimethoxy-flavanone-4′-O-[2″-O-(5‴-O-trans-cinnamoyl)-β-ᴅ-apiofuranosyl]-β-ᴅ-glucopyranoside (3), 2′-hydroxy-4′,6′-dimethoxy-chalcone-4-O-[2″-O-(5‴-O-trans-cinnamoyl)-β-ᴅ-apiofuranosyl]- β-ᴅ-glucopyranoside (4), 5,7-dimethoxy-flavanone-4′-O-[β-d-apiofuranosyl- (152)]-β-ᴅ-glucopyranoside (5). For the antinociceptive activity assessment the p-benzoquinone- induced writhing test and for the anti-inflammatory activity the carrageenan-induced hind paw edema model in mice were used. The ethyl acetate fraction in a dose of 250 mg/kg as well as compounds 2 and 5 in a 30 mg/kg dose were shown to possess remarkable antinociceptive and anti-inflammatory activities per os without inducing any apparent acute toxicity as well as gastric damage
A New Active Compound from Centaurea Species
Authors: Dumlu Melek U. , Gürkan Elcin
Published: February 2015

The extract containing sesquiterpene lactones of Centaurea iberica (Asteraceae) isolated was separated and a steroidal compound, which is stigmast-1,5-dien-3β-ol, was purified. The chemical structure was established based on spectroscopic data (UV, IR, MS, 1H NMR, 13C NMR). Both the extract and the compound showed significant antioxidant and antimicrobial activities
Molluscicidal Activity and New Flavonoids from Egyptian Iris germanica L. (var. alba)
Authors: Singaba Abdel Nasser B. , Ahmed Amer H. , Sinkkonen Jari , Ovcharenko Vladimir , Pihlaja Kalevi
Published: February 2015

The molluscicidal activity of leaf and rhizome extracts of Iris germanica L. (var. alba) against Biomphalaria alexandrina snails was evaluated and the rhizome extracts were found to be the most potent. Activity-guided fractionation revealed that the chloroform extract showed the highest molluscicidal activity (LC90 = 1.26 mg/l) among the tested extracts of the rhizomes. Fraction B prepared from the chloroform extract was the most potent molluscicide (LC90 = 0.96 mg/l) in addition, it showed a significant heart rate reduction in the snail after a 6- to 24-h exposure period. It also displayed a significant level of cercaricidal potential in a time-concentration relationship pattern. Chromatographic fractionation and purification of fraction B resulted in the isolation of two novel compounds: 5,2′-dihydroxy-3-methoxy-6,7- methylenedioxyflavone and 5,7,2′-trihydroxy-6-methoxyflavanone. Their structures were established by one- and two-dimensional NMR methods and mass spectrometry
Pinostrobin – An Anti-Leukemic Flavonoid from Polygonum lapathifolium L. ssp. nodosum (Pers.) Dans
Authors: Smolarz Helena D. , Mendyk Ewaryst , Bogucka-Kocka Anna , Kockic Janusz
Published: February 2015

Aim of study: Search for plant compounds possessing anti-leukemic properties. Results: We have shown that 5-hydroxy-7-methoxy flavanone (pinostrobin) isolated from Polygonum lapathifolium ssp. nodosum quickly penetrates through cytoplasm to the cellular nucleus of the cultured cells, and gives intensive apoptotic response in stimulating leukemic cells in vitro. The number of apoptotic cells increased with the concentration of pinostrobin: 10 nm D 25% and 60%; 100 nm D 45% and 76%; 1 μm D 70% and 88% for Jurkat and HL60 cell lines, respectively.

Conclusion: Pinostrobin may be considered as a good candidate for a leukemia chemopreventic agent
Herbicidal Potential of Catechol as an Allelochemical
Authors: Topal Süleyman , Kocaçalışkana Ismail , Arslan Orhan
Published: February 2015

Catechol is an allelochemical which belongs to phenolic compounds synthesized in plants. Its herbicidal effects on weed species; field poppy (Papaver rhoeas), creeping thistle (Cirsium arvense), henbit (Lamium amplexicaule) and wild mustard (Sinapis arvensis) were investigated using wheat (Triticum vulgare) and barley (Hordeum vulgare) species as control plants. In comparison to 2,4-D (a common synthetic herbicide), 13.64 mᴍ of catechol have been found to have a strong herbicidal effect, as effective as 2,4-D on field poppy weed by killing it, and a suppressive herbicidal effect on the other weeds by inhibiting their growth significantly. Concerning all the weeds, in general, elongation of the shoot was affected more negatively than that of the root. Fresh weights of the weeds were decreased by catechol significantly only in field poppy but not in other weeds. The study reveals that catechol is a potent inhibitor of growth of the weeds and therefore it can be evaluated as a herbicide for future weed management strategies
Cucurbitane Glucosides from Momordica charantia Leaves as Oviposition Deterrents to the Leafminer, Liriomyza trifolii
Authors: Mekuria Daniel Bisrat , Kashiwagi Takehiro , Tebayashi Shin-ichi , Kim Chul-Sa
Published: February 2015

The American serpentine leaf mining fly, Liriomyza trifolii, whose larva feeds on more than 120 plant species is well characterized by its high degree of polyphagy. Observations on the oviposition behavior by L. trifolii demonstrated that among cucurbitaceous plants, Momordica charantia is rarely attacked by L. trifolii. The methanol extract of M. charantia leaves strongly deterred the females from ovipositing on kidney bean leaves treated at a concentration of 1 g leaf equivalent extract/ml. Analysis of the polar fraction of the methanol extract of M. charantia leaves resulted in the isolation of a novel cucurbitane glucoside, 7-O- β-ᴅ-glucopyranosyl-3,23-dihydroxycucurbita-5,24-dien-19-al, named momordicine IV, along with another known compound, momordicine II. Momordicine II and IV deterred oviposition by L. trifolii significantly when bioassays were carried out on kidney bean leaves treated at 75.6 and 20.3 μg/cm2 leaf surface, respectively. There was no synergistic effect on oviposition deterrent when the two compounds were combined in their natural abundance
Production of Biosurfactant Using Different Hydrocarbons by Pseudomonas aeruginosa EBN-8 Mutant
Authors: Raza Zulfiqar Ali , Khan Muhammad Saleem , Khalid Zafar M. , Rehman Asma
Published: February 2015

The present investigation dealt with the use of previously isolated and studied gamma-ray mutant strain Pseudomonas aeruginosa EBN-8 for the production of biosurfactant by using different hydrocarbon substrates viz. n-hexadecane, paraffin oil and kerosene oil, provided in minimal medium, as the sole carbon and energy sources. The batch experiments were conducted in 250 mL Erlenmeyer flasks, containing 50 mL minimal salt media supplemented with 1% (w/v) hydrocarbon substrate, inoculated by EBN-8 and incubated at 37 °C and 100 rpm in an orbital shaker. The sampling was done on 24 h basis for 10 d. The surface tension of cell-free culture broth decreased from 53 to 29 mN/m after 3 and 4 d of incubation when the carbon sources were paraffin oil and n-hexadecane, respectively. The largest reduction in interfacial tension from 26 to 0.4 mN/m was observed with n-hexadecane, while critical micelle dilution was obtained as 50 X CMC for paraffin oil as carbon source. When grown on n-hexadecane and paraffin oil, the EBN-8 mutant strain gave 4.1 and 6.3 g of the rhamnolipids/ L, respectively. These surface-active substances subsequently allowed the hydrocarbon substrates to disperse readily as emulsion in aqueous phase
Phytochemical Differences between Calia secundiflora (Leguminosae) Growing at Two Sites in Mexico
Authors: Zavala-Cháveza Fernando , García-Mateos Rosario , Soto-Hernández Marcos , Kited Geofrey
Published: February 2015

The ecology and quinolizidine alkaloid chemistry of Calia secundiflora (Ortega) Yakovlev growing at two sites in Mexico were compared. At one site (Hidalgo) the vegetation was dominated by Flourensia resinosa and C. secundiflora, at the other site (Queretaro) C. secundiflora and Dodanaea viscosa were dominant. The Hidalgo site had shallower soils with less organic matter, N, P, and CaCO3. Seeds of C. secundiflora from each site accumulated a similar range of quinolizidine alkaloids, but the profile of alkaloids in the leaves and roots were different. The leaves and roots of plants at Hidalgo accumulated a similar range of alkaloids to the seeds with cytisine and/or N-methylcytisine being most abundant, whereas at Queretaro the leaves and roots accumulated lupinine, with other alkaloids being relatively minor constituents. The latter profile has not been reported previously for C. secundiflora
Sophoraflavanone G from Sophora pachycarpa Enhanced the Antibacterial Activity of Gentamycin against Staphylococcus aureus
Authors: Fakhimi Ali , Iranshahi Mehrdad , Emami Seyed Ahmad , Amin-Ar-Ramimeh Esam , Zarrini Gholamreza , Shahverdi Ahmad Reza
Published: February 2015

In this study the enhancement effect of Sophora pachy- carpa roots’ acetone extract on the antibacterial activity of gentamycin was evaluated against Staphylococcus au­reus. Disc diffusion and broth dilution methods were used to determine the antibacterial activity of gentamy­cin in the absence and presence of plant extract and its various fractions separated by TLC. A clinical isolate of S. aureus was used as test strain. The active component of the plant extract involved in enhancement of genta- mycin’s activity had Rf = 0.72 on a TLC plate. The spec­tral data (1H NMR. 13CNMR) of this compound re­vealed that this compound was 5,7,2′,4′-tetrahydroxy-8- lavandulylflavanone (sophoraflavanone G), previously isolated from Sophora exigua. In the presence of 0.03 μg/ mL of sophoraflavanone G the MIC value of gentamy­cin for S. aureus decreased from 32 to 8 μg/mL (a four­fold decrease). These results signify that the ultra-low concentration of sophoraflavanone G potentiates the an­timicrobial action of gentamycin suggesting a possible utilization of this compound in combination therapy against Staphylococcus aureus.
Transformations of Steroid Esters by Fusarium culmorum
Authors: Świzdor Alina , Kołek Teresa , Szpineter Anna
Published: February 2015

Keywords

The course of transformations of the pharmacological steroids: testosterone propionate, 4-chlorotestosterone acetate, 17β-estradiol diacetate and their parent alcohols in Fusarium culmorum AM282 culture was compared. The results show that this microorganism is capable of regioselective hydrolysis of ester bonds. Only 4-ene-3-oxo steroid esters were hydrolyzed at C-17. 17β-Estradiol diacetate underwent regioselective hydrolysis at C-3 and as a result, estrone - the main metabolite of estradiol - was absent in the reaction mixture. The alcohols resulting from the hydrolysis underwent oxidation at C-17 and hydroxylation. The same products (6β- and 15α-hydroxy derivatives) as from testosterone were formed by transformation of testosterone propionate, but the quantitative composition of the mixtures obtained after transformations of both substrates showed differences. The 15α-hydroxy deriv­atives were obtained from the ester in considerably higher yield than from the parent alcohol. The presence of the chlorine atom at C-4 markedly reduced 17β-saponification in 4-chloro- testosterone acetate. Only 3β,15α-dihydroxy-4α-chloro-5α-androstan-17-one (the main prod­uct of transformation of 4-chlorotestosterone) was identified in the reaction mixture. 6β- Hydroxy-4-chloroandrostenedione, which was formed from 4-chlorotestosterone, was |not de­tected in the extract obtained after conversion of its ester.
Purification and Amino Acid Sequence of Fructose-1,6-bisphosphate Aldolase from the Electric Organ of Electrophorus electricus (L.)
Authors: De-Simone Salvatore G. , Salles Christiane M. Cardoso de , Silva Celia M. Batistae , Hassón-Voloch Aida
Published: February 2015

A soluble fructose-1,6-bisphosphate aldolase enzyme has been purified 50.2-fold (2.36%) at the homogeneity from the electric organ of Electrophorus electricus by one step of DEAE- 52 anion exchange chromatography followed by Superose-12 gel filtration-FPLC. Like other aldolase enzymes the E. electricus protein is a dimer with two identical subunits of 45 kDa. The N-terminal (20 residues) revealed a high homology with S. aurata (75%, goldfish), R. ratus and M. musculus (mouse, 80%) enzymes.
Antimicrobial Activity of Six Constituents of Essential Oil from Salvia
Authors: Sonboli Ali , Babakhani Babak , Mehrabian Ahmad Reza
Published: February 2015

The antimicrobial activity of three Salvia species, i.e. S. santolinifolia, S. hydrangea and S. mirzayanii, essential oils were investigated. The essential oils were obtained from the aerial parts of plants and analyzed by GC-MS. The main constituents of aforementioned species were α-pinene (72.4%), β-pinene (6.6%) and limonene (5.3%); β-caryophyllene (25.1%), 1,8-cineol (15.2%) and caryophyllene oxide (11.5%); α-terpinenyl acetate (22.6%), 1,8-cineol (21.2%) and linalool (8.9%), respectively. Bioassays exhibited that the property of the oil of S. myrzayanii was superior to others. The antimicrobial activity of essential oil from Salvia species may well be due to the presence of synergy between six tested compounds (linalool, 1,8-cineol, α-pinene, β-pinene, β-caryophyllene and limonene) and other constituents of the oils with various degrees of antimicrobial activity. Among these, linalool and 1,8-cineol had the highest antimicrobial activity.
Triterpenic Acids and Flavonoids from Satureja parvifolia. Evaluation of their Antiprotozoal Activity
Authors: Baren Catalina van , Anao Ivie , Lira Paola Di Leo , Debenedetti Silvia , Houghton Peter , Croft Simon , Martino Virginia
Published: February 2015

Bioassay-guided fractionation of a Satureja parvifolia MeOH extract led to the isolation of eriodictyol, luteolin and ursolic and oleanolic acids as its active components against Plasmodium falciparum K1. This is the first time these compounds are reported as constituents of S. parvifolia. Ursolic acid showed an IC50 of 4.9 μg/ml, luteolin 6.4 μg/ml, oleanolic acid 9.3 μg/ml and eriodictyol 17.2 μg/ml. Antiplasmodial activity of eriodictyol and luteolin is reported here for the first time.

Besides, the four compounds showed activity against P. falciparum 3D7 strain and Trypanosoma brucei rhodesiense. Eriodictyol showed moderate activity on all the parasites but was the most selective compound as a result of its rather low cytotoxicity (IC50 174.2 μg/ml) on the mammalian KB cell line.
Insecticidal Activity of Synthetic Amides on Spodoptera frugiperda
Authors: Batista-Pereira Luciane G , Castral Thais C. , Silva Marina T. M. da , Amaral Bruna R. , Fernandes João B. , Vieira Paulo C. , Silva M. Fatima G. F. da , Corrêa Arlene G.
Published: February 2015

The phytochemistry of the genus Piper (Piperaceae) has been widely studied due to the biological properties of amides from these plants. In this work, we have synthesized and evaluated the toxic effect of 11 amides against the fall armyworm Spodoptera frugiperda larvae. The naturally occurring piperine was also evaluated. The most active amide was N- [3-(3′,4′-methylenedioxyphenyl)-2-(E)-propenoyl]piperidine with a LD50 of 1.07 μg mg−1 larvae. This amide was also evaluated by ingestion.
Thermostability and Photostability of Photosystem II of the Resurrection Plant Haberlea rhodopensis Studied by Chlorophyll Fluorescence
Authors: Georgieva Katya , Maslenkova Liliana
Published: February 2015

The stability of PSII in leaves of the resurrection plant Haberlea rhodopensis to high temperature and high light intensities was studied by means of chlorophyll fluorescence measurements. The photochemical efficiency of PSII in well-hydrated Haberlea leaves was not significantly influenced by temperatures up to 40 °C. F0 reached a maximum at 50 °C, which is connected with blocking of electron transport in reaction center II. The intrinsic efficiency of PSII photochemistry, monitored as Fv/Fm was less vulnerable to heat stress than the quantum yield of PSII electron transport under illumination (ΦPSII). The reduction of ΦPSII values was mainly due to a decrease in the proportion of open PSII centers (qP). Haberlea rhodopensis was very sensitive to photoinhibition. The light intensity of 120 μmol m−2 s−1 sharply decreased the quantum yield of PSII photochemistry and it was almost fully inhibited at 350 μmol m−2 s−1. As could be expected decreased photochemical efficiency of PSII was accompanied by increased proportion of thermal energy dissipation, which is considered as a protective effect regulating the light energy distribution in PSII. When differentiating between the three components of qN it was evident that the energy-dependent quenching, qE, was prevailing over photoinhibitory quenching, qI, and the quenching related to state 1-state 2 transitions, qT, at all light intensities at 25 °C. However, the qE values declined with increasing temperature and light intensities. The qI was higher than qE at 40 °C and it was the major part of qN at 45 °C, indicating a progressing photoinhibition of the photosynthetic apparatus.
Kinetics of Total Enzymatic Hydrolysis of Acetylcholine and Acetylthiocholine
Authors: Zdražilová Pavla , Štěpánkova Šárka , Vránova Martina , Komers Karel , Komersová Alena , Čegan Alexander
Published: February 2015

Kinetics and the mechanism of total in vitro hydrolyses (i.e. up to the exhaustion of substrate) of acetylcholine and acetylthiocholine by acetylcholinesterase and butyrylcholinesterase were studied in vitro in a batch reactor at 25 °C, pH 8 and ionic strength of 0.11 ᴍ. Every hydrolysis was monitored by 2 - 3 independent analytical methods. All studied types of enzymatic hydrolyses fulfilled the Michaelis - Menten reaction scheme with the irreversible second step. A table of obtained average values of rate constants and estimations of initial molar enzyme concentrations, and discussion of the results are presented.
Comparison of an Indirect Format ELISA on Modified Graphite and Polystyrene Surfaces against Triazines
Authors: Fouad Dina , Sound Iris , Holthues Heike , Pfeifer-Fukumura Ursula , Hammam Abbas M. , Ibrahim Said A. , Baumann Wolfram
Published: February 2015

An indirect format enzyme-linked immuno-sorbent assay (ELISA) on graphite rods (Ø 0.8 mm x 20 mm) and, for comparison, on microtiter plates has been developed against terbuthylazine. For this purpose, a series of 2-aminoalkyl-4-chloro-6-terbuthyl-s-triazine-2,6- diamine ELISA haptens with alkyl spacer lengths of 2, 4, 6, and 8 CH2 groups has been synthesized. The graphite rods or the microtiter plates were covered by a polymerized glutaraldehyde network, and the ELISA haptens have been coupled by imino coupling to the free aldehyde groups of that network. ε-Aminocaproic acid has been used as an agent to block unspecific binding sites. The ELISA was run in a competitive mode, where the immobilized ELISA hapten and the solute terbuthylazine as a target analyte compete for the solute antibody.
The Influence of Cholesterol on the Interaction between N-Dodecyl-N,Ndimethyl- N-benzylammonium Halides and Phosphatidylcholine Bilayers
Authors: Różycka-Roszak Bożenna , Przyczyna Adriana , Misiak Paweł , Pruchnik Hanna
Published: February 2015

Effects of N-dodecyl-N,N-dimethyl-N-benzylammonium halides (DBeAX) on thermotropic phase behavior of phosphatidylcholine/cholesterol bilayers as well as on 1H NMR spectra were studied. The surfactants were added either to the water phase or directly to the lipid phase (a mixed film was formed). The benzyl group, opposite to liposomes without cholesterol, is not incorporated into the bilayer in the gel state but only in the liquid state. All the halides DBeAX (particularly the chloride DBeAC) showed greater ability to destabilize the membrane structure in the presence than in the absence of cholesterol. The interaction of DBeAX with DPPC/cholesterol bilayers and subsequent changes in the phospholipid bilayer organization depended on the kind of counterion. The strongest effects were observed for chloride (most electronegative ion) and for iodide (largest ion). The effects of chloride and bromide on phase transition and 1H NMR spectra in the presence and absence of cholesterol were opposite. This is discussed in terms of the influence of counterions on the pair cholesterol-DPPC interactions.
Isolation and Characterization of Two Verrucarins from Myrothecium roridum
Authors: Schoettler Sylvia , Bascope Marcelo , Sterner Olov , Anke Timm
Published: February 2015

In the course of a screening for compounds inhibiting the growth of two human breast cancer cells lines, two highly cytotoxic compounds were isolated from fermentations of Myrothecium roridum. The elucidation of their structures revealed that they are macrocyclic trichothecenes of the verrucarin type, 16-hydroxyverrucarin A (1), reported here as a natural product for the first time, and verrucarin X (2), a new compound. Both 1 and 2 exhibit moderate antifungal activity and pronounced cytotoxic activity, with IC50 values in the nanomolar (1) and micromolar (2) range. Both compounds preferentially inhibit in vivo protein biosynthesis.
Antimicrobial Activity of Extracts of the Lichen Xanthoparmelia pokornyi and its Gyrophoric and Stenosporic Acid Constituents
Authors: Candan Mehmet , Yılmaz Meral , Tay Turgay , Kıvança Merih , Türk Hayrettin
Published: February 2015

The antimicrobial activity of the diethyl ether, acetone, chloroform, petroleum ether, and ethanol extracts of the lichen Xanthoparmelia pokornyi and its gyrophoric acid and stenosporic acid constituents has been screened against some foodborne bacteria and fungi. Both the extracts and the acids showed antimicrobial activity against Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Proteus vulgaris, Staphylococcus aureus, Streptococcus faecalis, Yersinia enterocolitica, Candida albicans and Candida glabrata. The extracts were inactive against the tested filamentous fungi. The MIC values of the extracts and the acids for the bacteria have also been determined.
A Comparison of Cell Wall Disruption Techniques for the Isolation of Intracellular Metabolites from Pleurotus and Lepista sp.
Authors: Taskova Rilka M. , Zorn Holger , Krings Ulrich , Bouws Henning , Berger Ralf G.
Published: February 2015

Different techniques were compared for their effectiveness in the disruption of the rigid cell walls of Basidiomycetes. Grinding under liquid nitrogen, stirred glass bead milling and enzymatic cell lysis were applied to the mycelia of Pleurotus sapidus and Lepista irina grown submerged. Each of the disruption procedures was evaluated by testing the quantity and quality of released intracellular metabolites: DNA, RNA, enzymes, and secondary metabolites. The most suitable method for nucleic acid isolation was grinding under liquid nitrogen, while bead mill homogenization was the superior technique for isolation of active enzymes. A new effective method is proposed for isolation of secondary metabolites with the aid of bead milling of fungal mycelia.
Element Content (Cu, Fe, Mn, Ni, Pb, and Zn) of the Ruderal Plant Verbascum olympicum Boiss. from East Mediterranean
Authors: Güleryüz Gürcan , Arslan Hülya , İzgi Belgin , Güçer Şeref
Published: February 2015

In this study, heavy metal content (Cu, Fe, Mn, Ni, Pb, Zn) was determined in soils and different organs of Verbascum olympicum Boiss. This species is endemic to Uludağ and spreads on destroyed areas such as: roadsides, developed building areas, ski lift stations and sheep folds. Soils and different organs (roots, stems, leaves and flowers) of plant samples were analyzed using an atomic absorption spectrophotometer for determining the element content. Heavy metal contents in soils and different organs in this species were highly correlated (P < 0.05). However, the contribution of plant organs to the accumulation capacity varied according to the metal. These results suggest that this species may be useful as a bioindicator for heavy metals.
Development of Visible Markers for Transgenic Plants and their Availability for Environmental Risk Assessment
Authors: Tamaoki Masanori , Imai Hiroe , Takahashi Hayato , Toda Yumio , Niwa Yasuo , Nakajima Nobuyoshi , Aono Mitsuko , Kubo Akihiro , Saji Hikaru
Published: February 2015

Monitoring of transgenic plants in the field is important, but risk assessment has entailed laborious use of invisible marker genes. Here, we assessed three easily visible marker transgenes - green fluorescent protein (GFP), R, and Nicotiana tabacum homeobox (NTH) 15 genes - for their potential use as marker genes for monitoring genetically modified plants. Transgenic Arabidopsis thaliana plants for each of these genes were visibly distinguished from wild-type plants. We determined the germination rate, 3-week fresh weight, time to first flowering, and seed weight of the transgenic plants to evaluate whether the expression of these marker genes affected the growth of the host. Introduction of GFP gene had no effect on the evaluated parameters, and we then used the GFP gene as a marker to assess the outcrossing frequency between transgenic and two Arabidopsis species. Our results showed that the hybridization frequency between transgenic plants and Arabidopsis thaliana was 0.24%, and between transformants and Arabidopsis lyrata it was 2.6% under experimental condition. Out-crossing frequency was decreased by extending the distance between two kinds of plants. Thus, the GFP gene is a useful marker for assessing the whereabouts of transgenes/transformants in the field. We also demonstrated that the GFP gene is possibly applicable as a selection marker in the process of generation of transgenic plants.
Does DNA Methylation Pattern Mark Generative Development in Winter Rape?
Authors: Filek Maria , Janiak Agnieszka , Szarejko Iwona , Grabczyńska Jadwiga , Macháčková Ivana , Krekule Jan
Published: February 2015

In this paper we report on changes in DNA methylation pattern in rape apices and leaves during transition from vegetative to reproductive stage due to grafting and/or vernalization. Grafted plants of winter rape (Brassica napus L., var. “Górczański”) (stock from vernalized, scion from non-vernalized plants) were used together with vernalized non-grafted plants. In addition, methylation status was determined also in spring rape (var. “Młochowski”) grown under normal and low temperature. The methylation-sensitive amplification polymorphism (MSAP) method with EcoRI/MspI and EcoRII/HpaII restriction enzymes was employed. The majority (ca. 68%) of analyzed loci (566 in winter and 551 in spring rape) were monomorphic, i.e. did not undergo methylation. Both cultivars showed a similar degree of methylation. 188 loci in winter and 176 in spring cultivars expressed changes in the methylation pattern. All differentially amplified fragments resulted from either full methylation of an internal cytosine or from hemi-methylation of an external cytosine. A pair-wise comparison showed that a similar number of loci underwent development-related methylation changes in apices of the winter and spring rape. The majority (80%) of changes were demethylation events in generative (vernalized) apices of the winter cultivar. However, an increased number of demethylated loci was detected in vernalized apices in comparison with generative, nonvernalized ones. In apices of vegetative and generative grafted plants the same number of demethylation events was observed. Overall, 10 MSAP loci were detected that expressed methylation changes in vernalized apices only; among them 7 loci underwent demethylation after vernalization and remained methylated in both vegetative and generative non-vernalized stage. Only 1 locus was demethylated in generative non-vernalized apices. Thus, most of demethylation events can be ascribed to vernalization and not to the generative stage. In leaves of winter rape methylation and demethylation events occurred with similar frequency, while in the spring cultivar more demethylation events were detected. The results show that during vernalization and transition to the generative stage different sets of genes are activated.
Characterization and Biocontrol Ability of Fusion Chitinase in Escherichia coli Carrying Chitinase cDNA from Trichothecium roseum
Authors: Pan Hongyu , Wei Yi , Xin Furong , Zhou Mingguo , Zhang Shihong
Published: February 2015

The antifungal mechanism of mycoparasitic fungi involves fungal cell wall degrading enzymes such as chitinases. Trichothecium roseum is an important mycoparasitic fungus with significant antifungal ability, but studies on chitinases of T. roseum were poor. Here, we report a novel chitinase cDNA isolated from T. roseum by PCR amplification based on conserved chitinase sequences. Southern blot analysis suggested that a single copy of the gene exists in the genome of T. roseum. The deduced open reading frame of 1,143 nucleotides encodes a protein of 380 amino acids with a calculated molecular weight of 41.6 kDa. The fusion chitinase expressed in Escherichia coli has been purified by single-step chromatography. It has a pI of pH 5.4 and expresses a thermal stability, but is insensitive to pH in a broad pH range. According to expectation, E. coli efficiently yielded a high amount of active chitinase. Remarkably, the fusion chitinase offered high antifungal activity.
Assessment of Genetic Diversity in 31 Species of Mangroves and their Associates through RAPD and AFLP Markers
Authors: Mukherjee Arup Kumar , Acharya Laxmikanta , Panda Pratap Chandra , Mohapatra Trilochan
Published: February 2015

Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers were used to assess the genetic diversity in 31 species of mangroves and mangrove associates. Four AFLP primer combinations resulted in the amplification of 840 bands with an average of 210 bands per primer combination and 11 RAPD primers yielded 319 bands with an average of 29 bands per primer. The percentage of polymorphism detected was too high indicating the high degree of genetic variability in mangrove taxa both at inter- and intra-generic levels. In the dendrogram, species belonging to a particular family/ genus, taxa inhabiting similar habitats or having similar adaptations tended to be together. There were exceptions too; as many unrelated species of mangroves form clusters. The intrafamilial classification and inter-relationships of genera in the family Rhizophoraceae could be confirmed by molecular analysis. Both the markers RAPD and AFLP were found equally informative and useful for a better understanding of the genetic variability and genome relationships among mangroves and their associated species.
The Effect of Trifolium, Raphanus, and Cistus Pollen Grains on Some Blood Parameters and Mesentery Mast Cells
Authors: Kolankaya Dürdane , Şentürkb Hakan , Tüylü Aslı Özkök , Hayretdağ Sibel , Selmanoğlua Güldeniz , Sorkun Kadriye
Published: February 2015

Three kinds of pollen taxa belonging to 3 families (Fabaceae - Trifolium spp., Brassicaceae - Raphanus spp. and Cistaceae - Cistus spp.) and commonly collected by honeybees were fed to mature male rats separately, in the form of 60 mg/animal/day for a 30-day period. The objective of this study was to investigate any positive effects or possible side effects of the use of pollen on the immune system. This was achieved through blood analysis and cell count on blood, hemoglobin, erythrocyte and immune system cells. The cell concentration of mast cells, degranulization and cell localization were investigated in prepared mesentery tissue samples. Histological investigations of the stomach and duedenum sections of pollen-fed rats were carried out to learn the reason for eosinophil gastroenteritis in the alimentary canal.

The eosinophil and lymphocyte levels of rats fed with pollen of Trifolium spp., Raphanus spp., and Cistus spp. were observed to have increased blood cell counts, while neutrophil and monocyte levels decreased; different values were found in basophil leucocytes between the pollen groups. Differing reductions in mesentery mast cell concentration, degranulization and cell localization were found. Within the three separate pollens, the rats having been fed with Cistus spp. pollen were observed to have higher blood lymphocyte, eosinophil, hemoglobin and hematocrit values than those fed with the others, as well as low mesentery mast cell concentration. Hemoglobin values were determined to increase at a proportion of between 10.0-11.3%. No difference was found in other blood parameters.

The fat proportion of the male rats fed with the three taxa was between 4.03-8.75%, while that for protein proportion was between 16.11-24.25%. Male rats receiving these taxa did not experience allergic reactions and it is possible to argue that the low protein and fat content of these pollens have a strengthening effect on the immune systems by the increase in lymphocyte content and the amount of hemoglobin leads to an increase of oxygen transport capacity in the tissues.
Monitoring of BHT-Quinone and BHT-CHO in the Gas of Capsules of Asclepias physocarpa
Authors: Ma Bing-Ji , Peng Hua , Liu Ji-Kai
Published: February 2015

Three volatile components, namely benzoic acid ethyl ester (1), 2,6-di-tert-butyl-p-benzoquinone (BHT-quinone) (2), and 3,5-di-tert-butyl-4-hydroxybenzaldehyde (BHT-CHO) (3), were detected from the gas in the capsules of Asclepias physocarpa by means of GC/MS analysis. BHT-quinone and BHT-CHO as organic pollutants are the degradation products of the antioxidant 2,6-ditert-butyl-4-methylphenol (BHT). Ground water, lake water and/or rain water are a source of BHT metabolites in the plant Asclepias physocarpa.
In vivo Analgesic and Anti-Inflammatory Activities of Ursolic Acid and Oleanoic Acid from Miconia albicans (Melastomataceae)
Authors: Vasconcelos Maria Anita L. , Royo Vanessa A. , Ferreira Daniele S. , Crotti Antonio E. Miller , e Silva Márcio L. Andrade , Carvalho José Carlos T. , Bastos Jairo Kenupp , Cunha Wilson R.
Published: February 2015

The aim of this work was to use in vivo models to evaluate the analgesic and anti-inflammatory activities of ursolic acid (UA) and oleanoic acid (OA), the major compounds isolated as an isomeric mixture from the crude methylene chloride extract of Miconia albicans aerial parts, in an attempt to clarify if these compounds are responsible for the analgesic properties displayed by this plant. Ursolic acid inhibited abdominal constriction in a dose-dependent manner, and the result obtained at a content of 40 mg kg-1 was similar to that produced by administration of acetylsalicylic acid at a content of 100 mg kg-1. Both acids reduced the number of paw licks in the second phase of the formalin test, and both of them displayed a significant anti-inflammatory effect at a content of 40 mg kg-1. It is noteworthy that the administration of the isolated mixture, containing 65% ursolic acid/35% oleanolic acid, did not display significant analgesic and anti-inflammatory activities. On the basis of the obtained results, considering that the mixture of UA and OA was poorly active, it is suggested that other compounds, rather than UA and OA, should be responsible for the evaluated activities in the crude extract, since the crude extract samples displayed good activities.
Cytotoxic and Proapoptotic Activity of Diterpenoids from in vitro Cultivated Salvia sclarea Roots. Studies on the Leukemia Cell Lines
Authors: Różalski Marek , Kuźma Łukasz , Wysokińska Halina , Krajewska Urszula
Published: February 2015

Four diterpenoids, ferruginol, salvipisone, aethiopinone and 1-oxoaethiopinone, were isolated from transformed roots of Salvia sclarea. Salvipisone and aethiopinone showed relatively high cytotoxicity against HL-60 and NALM-6 leukemia cells (IC50 range 0.6-7.7 μg/ mL which is equal to 2.0-24.7 μᴍ), whereas 1-oxoaethiopinone and ferruginol were less active in this regard. Moreover, we have found that all four diterpenoids of S. sclarea had equal cytotoxic activity against parental HL-60 and multidrug-resistant HL-60 ADR cells, what indicates that they are poor substrates for transport by multidrug resistance-associated protein (MRP1). Caspase-3 activity determinations showed that salvipisone and aethiopinone were able to induce apoptosis in a time- and concentration-dependent manner. The results obtained in this study show that S. sclarea diterpenoids aethiopinone and salvipisone may be useful in the treatment of human cancers, especially in the case of drug resistance.
Antimicrobial Activity of Extracts of Chemical Races of the Lichen Pseudevernia furfuracea and their Physodic Acid, Chloroatranorin, Atranorin, and Olivetoric Acid Constituents
Authors: Türk Hayrettin , Yılmaz Meral , Tay Turgay , Türk Ayşen Özdemir , Kıvanç Merih
Published: February 2015

The antimicrobial activity and the MIC values of the ethanol, chloroform, diethyl ether, and acetone extracts of the chemical races of Pseudevernia furfuracea (var. furfuracea and var. ceratea) and their physodic acid, chloroatranorin, atranorin, and olivetoric acid constituents have been investigated against some microorganisms. Nearly all extracts of both chemical races showed antimicrobial activity against Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Proteus vulgaris, Staphylococcus aureus, Streptococcus faecalis, Yersinia enterocolitica, Candida albicans, Candida glabrata, Alternaria alternata, Ascochyta rabiei, Aspergillus niger, Fusarium culmorum, Fusarium moniliforme, Fusarium oxysporum, Fusarium solani, and Penicillium notatum. There was no antimicrobial activity of the extracts against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Pseudomonas syringae, Salmonella typhimurium, Alternaria citri, Alternaria tenuissima, and Gaeumannomyces graminis. Chloroatranorin and olivetoric acid were active against the same microorganisms with few exceptions. Physodic acid was active against about the same bacteria and yeasts and inactive against all of the filamentous fungi tested. Also no activity of atranorin against the filamentous fungi was observed.
Secondary Metabolites and Insecticidal Activity of Anemone pavonina
Authors: Varitimidisa Christos , Petrakis Panos V. , Vagias Constantinos , Roussis Vassilios
Published: February 2015

The insecticidal properties of the crude extracts of the leaves and flowers of Anemone pavonina were evaluated on Pheidole pallidula ants and showed significant levels of activity. Bioassay-guided fractionations led to the isolation of the butenolide ranunculin (1) as the active principle. Chemical investigations of the extracts showed them to contain as major components the sitosterol glycopyranoside lipids 2-5 and the glycerides 6-8. The structures of the metabolites were elucidated, following acetylation and hydrolysis of the natural products, by interpretation of their NMR and mass spectral data. The uncommon lipid metabolites 2-8 were isolated for the first time from the genus Anemone and this is the first report of insecticidal activity of the Anemone metabolite ranunculin against ants.
Altered Toxicities of Fatty Acid Salts in Green Paramecia Cultured in Different Waters
Authors: Kadono Takashi , Uezu Kazuya , Kosaka Toshikazu , Kawano Tomonori
Published: February 2015

Detergents including fatty acid salts act as surface-active agents and thus possibly damage the plasma membrane structures of aquatic organisms. Therefore, when excess, the houseused and industrial outflows of such detergents into aquatic environments may have considerable impacts on the ecosystem. In this study, we propose the use of green paramecia (Paramecium bursaria) for assessing the acute toxicity of eight fatty acid salts (Na and K salts of oleate, palmitate, laurate and myristate) under various water conditions. The Paramecium in the stationary phase were used for a toxicity assay carried out on 12-well microplates and the median lethal concentration (LC50) was determined for each fatty acid salt. In the low mineral culture medium prepared with ultra-pure water, the LC50 for each fatty acid ranged from 5.8 to 144 ppm (w/v). The toxic levels of fatty acid salts differed in the following order: laurate, myristate ≥ oleate, palmitate. The toxic levels of oleate and palmitate salts were ca. 10-fold lower than those of laurate and myristate salts. When river water and local tap water instead of ultra-pure water were used for culturing, the toxic levels of all fatty acid salts were drastically lowered compared to the low mineral condition by 30- to 100-fold (198-660 ppm, w/v). Similar detoxification effect was observed when Ca or Mg was added to the low mineral culture media, indicating that the toxicity of fatty acid salts can be notably lowered as the mineral content increases. As we demonstrated that toxicities of fatty acid salts can be lowered in river water and tap water compared to the low mineral condition, some chemical substances behave differently in the different water conditions. Therefore, the use of natural waters reflecting the real environmental conditions in further collection of data on the ecotoxicity impacts of variety of chemicals is highly encouraged.