bluuebunny/crossref_metadata_2025 huggingface dataset:
Conserved and Non-Conserved Loci of the Glucagon Gene in Old World Ruminating Ungulates
Authors: Warda Mohamad , Gouda Eman M. , El-Behairy Adel M. , Mousa Said Z.
Published: February 2015

The homology and diversification of genomic sequence encoding glucagon gene among native Egyptian buffalos, camel and sheep were tested using cattle as model. Oligodeoxynucleotide primers designed from the available GenBank data were used for PCR probing of the glucagon gene encoding sequence at different loci. The DNA oligomer probes were constructed to flank either the whole gene encoding sequence or different intra-gene encoding sequences. The PCR products were visualized using agarose gel electrophoresis. All species showed a same size band of prepro-glucagon when PCR was used to amplify the whole gene encoding sequence. In contrary, amplifications of different intra-gene loci failed to give the same results. The results indicated variable degrees of diversity among old world ruminating ungulates in the glucagon gene encoding sequence. Compared with other ruminants, the variation appears predominantly in camel. Surprisingly, the similarity in size between both amplification products of whole gene encoding sequence and the proposed size of glucagon cDNA definitely excludes the possibility of large intervening introns spanning the genomic sequence of the glucagon gene in these species. This indicates that, in contrast to other tested mammals, the glucagon gene includes an essentially full-length copy of glucagon mRNA. The study revealed a possible new aspect of glucagon gene evolution in order to correlate its corresponding protein function among different ruminant species
Binding of an Oxindole Alkaloid from Uncaria tomentosa to Amyloid Protein (Aβ1-40)
Authors: Frąckowiak Teresa , Bączek Tomasz , Kaliszan Roman , Żbikowska Beata , Gleńsk Michał , Fecka Izabela , Cisowski Wojciech
Published: February 2015

The primary aim of this work was to determine the interactions of an oxindole alkaloid (mitraphylline) isolated from Uncaria tomentosa with β-amyloid 1-40 (Aβ1-40 protein) apply­ing the capillary electrophoresis (CE) method. Specifically the Hummel-Dreyer method and Scatchard analysis were performed to study the binding of oxindole alkaloids with Aβ1-40 protein. Prior to these studies extraction of the alkaloid of interest was carried out. Identifica­tion of the isolated alkaloid was performed by the use of thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) combined with electrospray ionization mass spectrometry (ESI-MS). The proposed approach was proved to be an efficient and accurate method for specific compound isolation and identification purposes. Moreover, ana­lytical information from the CE approach can be considered as the valuable tool for binding constant determination. The binding constant of mitraphylline with Aβ1-40 protein deter­mined by the Hummel-Dreyer method and Scatchard analysis equals K = 9.95 x 105 m-1. The results obtained showed the significant binding of the tested compound with Aβ1-40 protein. These results are discussed and interpreted in the view of developing a strategy for identifica­tion of novel compounds of great importance in Alzheimer disease therapy.
Ionic Liquids as Novel Solvent Additives to Separate Peptides
Authors: Bączek Tomasz , Sparzak Barbara
Published: February 2015

A novel analytical approach involving the addition of an ionic liquid into the mobile phase of the thin-layer chromatography (TLC) system during the optimization of chromatographic separation of peptides was demonstrated. Different behavior of peptides in the TLC sytem was observed after the addition of 1,3-dimethylimidazolium methyl sulfate to the eluent in comparison to the system without the ionic liquid. The objective of the work was to study the effect of the addition of different contents of ionic liquid to the mobile phase comprising mostly water and to observe the behavior of peptides’ retention. The potential usefulness of environmentally friendly ionic liquids for the optimization of separation of peptides was demonstrated. An increase of Rf values was observed with increasing the ionic liquid content in the mobile phase. The benefits of the used approach were related to the separation achieved. Finally, quantitative structure-retention relationships (QSRR) were used for the studies on the predictions of peptides’ retention in the TLC systems with the addition of ionic liquid in terms of the predictions performed recently in HPLC systems.
A β-Mannanase from Bacillus subtilis B36: Purification, Properties, Sequencing, Gene Cloning and Expression in Escherichia coli
Authors: Li Ya Nan , Meng Kun , Wang Ya Ru , Yao Bin
Published: February 2015

MANB36, a secrete endo-β-1,4-D-mannanase produced by Bacillus subtilis B36, was puri­fied to homogeneity from a culture supernatant and characterized. The optimum pH value for the mannanase activity of MANB36 is 6.4 and the optimum temperature is 50 °C. The enzyme activity of MANB36 is remarkably thermostable at 60 °C and the specific activity of MANB36 is 927.84 U/mg. Metal cations (except Hg2+ and Ag+), EDTA and 2-mercaptoetha- nol (2-ME) have no effects on enzyme activity. This enzyme exhibits high specificity with the substituted galactomannan locust bean gum (LBG). The gene encoding for MANB36, manB36, was cloned by PCR and sequenced. manB36 contains a single open reading frame (ORF) consisting of 1104 bp that encodes a protein of 367 amino acids. The predicted mo­lecular weight of 38.13 kDa, calculated by the deduced protein of the gene manB36 without signal peptide, coincides with the apparent molecular weight of 38.0 kDa of the purified MANB36 estimated by SDS-PAGE. The mature protein of MANB36 has been expressed in Escherichia coli BL21 and the expressed mannanase has normal bioactivity.
O-
2 Activates Leaf Injury, Ethylene and Salicylic Acid Synthesis, and the Expression of O3-Induced Genes in O3-Exposed Tobacco
Authors: Ogawa Daisuke , Nakajima Nobuyoshi , Tamaoki Masanori , Aono Mitsuko , Kubo Akihiro , Kamada Hiroshi , Saji Hikaru
Published: February 2015

O3 is the major component of photochemical oxidants and gives rise to visible injuries on plant leaves. In O3-exposed plants, O-
2 is produced before the formation of the injury, but the role that O-
2 plays in plant response to O3 exposure is still unknown. To clarify its role, we observed the behavior of plants during O3 exposure after pretreatment with tiron, which is an O-
2 scavenger. When tiron-pretreated tobacco cv. Bel W3 was exposed to O3, leaf damage was attenuated. In O3-exposed tobacco, tiron inhibited increases in the levels of ethylene and salicylic acid, which promote leaf injury. Tiron pretreatment also suppressed increases in the expression of O3-induced genes. These results suggest that O-
2 is involved in many plant responses induced by O3 exposure. Bel B, a tobacco cultivar that is genetically related to Bel W3, is reported to be more resistant to O3 than Bel W3, but the reason for this difference is unclear. We investigated the differences between the responses of Bel B and tiron-pretreated Bel W3 to O3 exposure, and we discuss the reasons for the resistance to O3 by comparing the phenotype of Bel B with that of tiron-pretreated Bel W3.
Imatinib (STI571) Inhibits DNA Repair in Human Leukemia Oncogenic Tyrosine Kinase-Expressing Cells
Authors: Majsterek Ireneusz , Arabski Michal , Czechowska Agnieszka , Pytel Dariusz , Morawiec Zbigniew , Morawiec-Bajda Alina , Blasiak Janusz
Published: February 2015

BCR/ABL oncogene, as a result of chromosome aberration t(9;22), is the pathogenic principle of almost 95% of human chronic myeloid leukemia (CML). Imatinib (STI571) is a highly selective inhibitor of BCR/ABL oncogenic tyrosine kinase used in leukemia treatment. It has been suggested that BCR/ABL may contribute to the resistance of leukemic cells to drug and radiation through stimulation of DNA repair in these cells. To evaluate further the influence of STI571 on DNA repair we studied the efficacy of this process in BCR/ABLpositive and -negative cells using single cell electrophoresis (comet assay). In our experiments, K562 human chronic myeloid leukemia cells expressing BCR/ABL and CCRF-CEM human acute lymphoblastic leukemia cells without BCR/ABL expression were employed. The cells were exposed for 1 h at 37 ∞C to N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) at 5 μᴍ, mitomycin C (MMC) at 50 μᴍ or to γ-radiation at 15 Gy with or without a 24 h preincubation at 1 μᴍ of STI571. The MTT cells survival after 4 days of culture showed that STI571 enhanced the cytotoxity of the examined compounds in the K562 line. Further it was found, that the inhibitor decreased the efficacy of DNA repair challenged by each agent, but only in the K562 expressing BCR/ABL. Due to the variety of DNA damage induced by the employed agents in this study we can speculate, that BCR/ABL may stimulate multiple pathways of DNA repair. These results extend our previous studies performed on BCR/ABLtransformed mouse cells onto human cells. It is shown that BCR/ABL stimulated DNA repair in human leukemia cells. In conclusion we report that STI571 was found to inhibit DNA repair and abrogate BCR/ABL-positive human leukemia cells therapeutic resistance.
Cytotoxic Abietane Diterpenes from Hyptis martiusii Benth.
Authors: Araújo Edigênia Cavalcante da Cruz , Lima Mary Anne S. , Montenegro Raquel C. , Nogueira Marcelle , Costa-Lotufo Letícia V. , Pessoa Cláudia , Moraes Manoel Odorico de , Silveira Edilberto R.
Published: February 2015

From roots of Hyptis martiusii Benth. two tanshinone diterpenes were isolated, the new 7β-hydroxy-11,14-dioxoabieta-8,12-diene (1) in addition to the known 7α-acetoxy-12-hydroxy- 11,14-dioxoabieta-8,12-diene (7α-acetoxyroyleanone) (2). Structures of 1 and 2 were established by spectroscopic means. The cytotoxic activity against five cancer cell lines was evaluated. Compounds 1 and 2 displayed considerable cytotoxic activity against several cancer cell lines with IC50 values in the range of 3.1 to 11.5 μg/ml and 0.9 to 7.6 μg/ml, respectively. The cytotoxic activity seemed to be related to inhibition of DNA synthesis, as revealed by the reduction of 5-bromo-2′-deoxyuridine incorporation and induction of apoptosis, as indicated by the acridine orange/ethidium bromide assay and morphological changes after 24 h of incubation in leukemic cells.
Phytoremediation of Pesticide Wastes in Soil
Authors: Henderson Keri L. D. , Belden Jason B. , Zhao Shaohan , Coats Joel R.
Published: February 2015

Soils at agrochemical dealer sites often are contaminated with pesticide residues from decades of accidental and incidental spillage. We have determined that prairie grasses native to the Midwestern U.S. are suitable for phytoremediation because they are tolerant of most herbicides and of climatic extremes, such as heat, cold, drought, and flooding. A mixed stand of big bluestem, switch grass, and yellow indiangrass develops a rhizosphere with microflora that can readily detoxify pesticide residues. Specific atrazine-degrading bacteria or the free enzyme atrazine chlorohydrolase also can enhance the rate of biotransformation of atrazine in soil. Metolachlor degradation can be accelerated significantly by the prairie grass/rhizosphere effect. Several grasses used in filter strips have also been evaluated for their pesticidedegradation capabilities. The prairie grasses also have been demonstrated to reduce the rates of leaching of pesticides through intact soil columns, since less water leaches out of vegetated soil columns compared to non-vegetated soil columns. The evaluation of the degree of success of remediation has relied heavily on chemical residue analysis, but recent studies on biological endpoints have shown promise for providing more ecologically relevant indications of the potential exposure of organisms to pesticides in the soil. Earthworm 8-day bioaccumulation assays and root growth assays have shown the value of assessing the bioavailability of the residues. Mass balance experiments have utilized radiolabeled atrazine and metolachlor to ascertain the complete metabolism and binding profile of those two pesticides in phytoremediation studies.
Protein Mobilization and Proteolytic Enzyme Activities during Seed Germination of Broad Bean (Vicia faba L.)
Authors: Kırmızı Serap , Güleryüz Gürcan
Published: February 2015

The protein mobilization from attached and detached seeds of Vicia faba L. cv. Eresen 87 (Fabaceae) was investigated. While the total soluble protein content decreased, the free amino acid content increased during the 7 days germination period. Among the three proteolytic enzymes, only endopeptidase activity was found to be affected by the removal of the embryonic axis. Leucine aminopeptidase activity was high at the beginning, then it decreased; carboxypeptidase activity reached the highest value at day 5. In order to examine the effects of plant growth regulators on detached cotyledons incubated with plant growth regulators [10−4 ᴍ benzyladenine (BA), gibberellic acid (GA3), indole acetic acid (IAA) and 10−5 ᴍ abscisic acid (ABA)], only benzyladenine was found promotive on protein mobilization. Our results suggest that the removal of the embryonic axis in seeds of Vicia faba L. cv. Eresen 87 decreases protein mobilization and endopeptidase activity.
Composition and Antimicrobial Activity of the Essential Oil of Dicyclophora persica Boiss. from Iran
Authors: Salehi Peyman , Sonboli Ali , Mohammadi Fatemeh
Published: February 2015

The chemical composition of the essential oil of Dicyclophora persica Boiss. was identified by GC and GC-MS analysis. The analysis of the oil resulted in the identification of forty-five components constituting 98.6% of the total oil. The main constituents were α-pinene (31.5%), (Z)-β-ocimene (23.3%), p-cymene (6.7%) and (E)-β-ocimene (5.4%). The antimicrobial activity of the oil was tested by the disk diffusion method against four Gram-positive (Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus faecalis) and three Gram-negative (Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) bacteria together with a fungus (Aspergillus niger). The oil showed strong inhibition activity toward all the tested microorganisms except for Pseudomonas aeruginosa.
Analysis of Essential Oil Composition of Thymbra spicata var. spicata: Antifungal, Antibacterial and Antimycobacterial Activities
Authors: Kılıç Turgut
Published: February 2015

The fresh leaves and brine of leaves of Thymbra spicata var. spicata (KARAKIZTM) were analyzed by hydrodistillation, headspace and GC/MS techniques. The main components were determined as carvacrol, p-cymene, β-myrcene, γ-terpinene, α-terpinene and trans-caryophyllene. The essential oil and the main compounds, carvacrol and trans-caryophyllene, have been tested against E. coli, S. epidermidis, B. subtilis, S. aureus, S. typhimurium, K. pneumoniae, P. aeruginosa, E. faecalis and C. albicans. While the essential oil and carvacrol showed strong activity against all microorganisms, except P. aeruginosa, trans-caryophyllene showed activity only against C. albicans. The essential oil and carvacrol also showed strong antimycobacterial activity.
Phytotoxicity of the Volatile Monoterpene Citronellal against Some Weeds
Authors: Singh Harminder Pal , Batish Daizy R. , Kaur Shalinder , Kohli Ravinder K. , Arora Komal
Published: February 2015

A study was undertaken to assess the phytotoxicity of citronellal, an oxygenated monoterpenoid with an aldehyde group, towards some weedy species [Ageratum conyzoides L., Chenopodium album L., Parthenium hysterophorus L., Malvastrum coromandelianum (L.), Garcke, Cassia occidentalis L. and Phalaris minor Retz.]. A significant effect on weed emergence and early seedling growth was observed in a dose-response based laboratory bioassay in a sand culture. Emergence of all test weeds was completely inhibited at 100 μg/g sand content of citronellal. Seeds of A. conyzoides and P. hysterophorus failed to emerge even at 50 μg/g content. Root length was inhibited more compared to shoot length. The failure of root growth was attributed to the effect of citronellal on the mitotic activity of growing root tips cells as ascertained by the onion root tip bioassay. At 2.5 mm treatment of citronellal, mitosis was completely suppressed and at higher concentrations cells showed various degrees of distortion and were even enucleated. The post-emergent application of citronellal also caused visible injury in the form of chlorosis and necrosis, leading to wilting and even death of test weeds. Among the test weeds, the effect was severe on C. album and P. hysterophorus. There was loss of chlorophyll pigment and reduction in cellular respiration upon citronellal treatment indicating the impairment of photosynthetic and respiratory metabolism. Scanning electron microscopic studies in C. occidentalis leaves upon treatment of citronellal revealed disruption of cuticular wax, clogging of stomata and shrinkage of epidermal cells at many places. There was a rapid electrolyte leakage in the leaf tissue upon exposure to citronellal during the initial few hours. In P. minor electrolyte leakage in response to 2 mm citronellal was closer to the maximum leakage that was obtained upon boiling the tissue. The rapid ion leakage is indicative of the severe effect of citronellal on the membrane structure and loss of membrane integrity. In all, the study concludes that citronellal causes a severe phytotoxicity on the weeds.
Sennosides A and B Production by Hairy Roots of Senna alata (L.) Roxb.
Authors: Putalun Waraporn , Pimmeuangkao Suwat , De-Eknamkul Wanchai , Tanaka Hiroyuki , Shoyama Yukihiro
Published: February 2015

Hairy roots of Senna alata transformed with Agrobacterium rhizogenes, strain ATCC 15834 were induced and grown in half-strength Murashige and Skoog (MS) medium. Effects of sucrose contents and hormones on the growth and sennosides A, B production were investigated. Hairy roots cultured on hormone-free half-strength MS medium containing 5% sucrose under dark condition mostly stimulated the growth of hairy roots and increased the content of sennosides A and B yielding (169 ± 4) and (34 ± 3) μg g-1 dry wt, respectively.
Germination Responses to GA3 and Stratification of Threatened Festuca L. Species from Eastern Mediterranean
Authors: Çelikler Serap , Güleryüz Gürcan , Bilaloğlu Rahmi
Published: February 2015

The seed germination characteristics of three threatened Festuca sp. [F. punctoria Sm., F. cyllenica Boiss. et Heldr. subsp. uluana Markgr.-Dannenb., F. paphlagonica (St.-Yves) Markgr.-Dannenb. subsp. paphlagonica] were investigated. These species are endemic and spread on alpine belt. The study was carried out with wet-cold and dry-cold stratification throughout 15 days, different doses of GA3 (50, 100 and 150 ppm) and hormone-stratification combined treatments, and non-treatment series. We found that the germination rates of three fescue seeds for various treatment series were different. The mean germination percentage of F. cyllenica was higher (80%) than that of F. punctoria and F. paphlagonica which were fairly low (50-60%). Germination rates increased by wet-stratification treatment in F. punctoria and also increased with 100 ppm GA3 application to the seeds of F. paphlagonica. When taken into consideration the germination percentages of all fescue species, the seeds of F. punctoria and F. paphlagonica can be dormant, but the seeds of F. cyllenica are non-dormant.
Phylogenetic Relations of Rhizoplaca Zopf. from Anatolia Inferred from ITS Sequence Data
Authors: Cansaran Demet , Aras Sümer , Kandemir İrfan , Halıcı Gökhan
Published: February 2015

Like many lichen-forming fungi, species of the genus Rhizoplaca have wide geographical distributions, but studies of their genetic variability are limited. The information about the ITS rDNA sequences of three species of Rhizoplaca from Anatolia was generated and aligned with other species from other countries and also with the data belonging to Lecanora species. The examined species were collected from the volcanic rocks of Mount Erciyes which is located in the middle of Anatolia (Turkey). The sequence data aligned with eight other samples of Rhizoplaca and six different species of Lecanora were obtained from GenBank. The results support the concept maintained by Arup and Grube (2000) that Rhizoplaca may not be a genus separate from Lecanora. According to the phylogenetic tree, Rhizoplaca melanopthalma from Turkey with two different samples of R. melanopthalma from Arizona (AF159929, AF159934) and a sample from Austria formed a group under the same branch. R. peltata and R. chrysoleuca samples from Anatolia located in two other branches of the tree formed sister groups with the samples of the same species from different countries. Although R. peltata remained on the same branch with other samples of the same species from other countries it was placed in a different branch within the group. When the three species from Anatolia were considered alone, it was noticed that Rhizoplaca melanopthalma and Rhizoplaca peltata are phylogenetically closer to each other than Rhizoplaca chrysoleuca; the morphological characteristics also support this result.
Host Selection in Tomicus piniperda L.: Composition of Monoterpene Hydrocarbons in Relation to Attack Frequency in the Shoot Feeding Phase
Authors: Almquist Ann-Charlotte , Fäldt Jenny , Yart Annie , Chevet Yohann , Sauvard Daniel , Lieutier Francois , Borg-Karlson Anna-Karin
Published: February 2015

The aim of this study was to investigate the host selection capacity of the pine shoot beetle, Tomicus piniperda, in the shoot-feeding phase and analyze the chiral and non-chiral host volatiles by means of GC-MS and 2D-GC in five Pinus species originating from France (Pinus sylvestris, P. halepensis, P. nigra laricio, P. pinaster maritima, P. pinaster mesogeensis). Dominating monoterpenes were (-)-α-pinene, (+)-α-pinene, (-)-β-pinene and (+)-3-carene. The amounts of the enantiomers varied considerably within and among the species. In a principal component analysis-plot, based on the absolute amounts of 18 monoterpene hydrocarbons, separation of the pine species into two groups was obtained. P. halepensis and P. sylvestris were grouped according to the amount of (+)-α-pinene and (+)-3-carene, while P. nigra laricio, P. pinaster maritima and P. pinaster mesogeensis were grouped according to (-)-α-pinene and (D)-β-pinene. P. nigra laricio was the species most attacked and P. halepensis the one least attacked by T. piniperda.
Catalytic Efficiencies of Alkaline Proteinases from Microorganisms
Authors: Georgieva Dessislava , Genov Nicolay , Voelter Wolfgang , Betzel Christian
Published: February 2015

Catalytic efficiencies of proteinase K and mesentericopeptidase were determined using series of peptide-4-nitroanilide substrates and compared with those of subtilisin DY, savinase and esperase. For each enzyme the subsites S1-S4 were characterized. The data for the enzyme specificities were related to our high resolution X-ray models of the five enzymes and their complexes with peptides. The catalytic efficiencies of the alkaline proteinases are modulated by the hydrophobicity, solvent accessibility, flexibility and electrostatic effects in the substrate binding sites. The longer and nonpolar S1 loop offers more possibilities for hydrophobic interactions and increases the enzyme efficiency. S2 is a small narrow cleft which limits the possibilities for effective substitutions in P2. The wide specificity of S3 is due to its location on the protein surface of all investigated proteinases. The affinity of S4 for aromatic groups depends on the nature of the residues building the hydrophobic cavity.
Antimicrobial Compounds from Coleonema album (Rutaceae)
Authors: Esterhuizen Lindy L. , Meyer Riaan , Dubery Ian A.
Published: February 2015

Coleonema album, a member of the South African fynbos biome, was evaluated for its antimicrobial activity associated with its secondary metabolites. Ethanol- and acetone-based extracts obtained from plants from two different geographical areas were analyzed. A bioassay- guided fractionation methodology was followed for rapid and effective screening for the presence of bioactive compounds. The TLC-bioautographic method, used to screen the plant extracts for antimicrobial activity and localization of the active compounds, indicated the presence of a number of inhibitory compounds with activity against the microorganisms (E. coli, B. subtilis, E. faecalis, P. aeruginosa, S. aureus, M. smegmatis, M. tuberculosis, C. albicans, C. cucumerinum) tested. Evaluation of the inhibitory strength of each extract by the serial microdilution assay indicated that the C. album extracts inhibited effectively all the microorganisms, with the minimum inhibitory concentrations in the low mg ml-1 range. Identification and structural information of the bioactive components were obtained by a combination of preparative TLC and LC-MS. It revealed the presence of coumarin aglycones which were responsible for the observed antimicrobial activities. The results of this study indicate that C. album possesses strong antimicrobial activity against a wide range of microorganisms that warrants further investigation into the use of the extracts or their active constituents as a potential source for novel drugs.
Biologically Active Constituents of a Polyphenol Extract from Geranium sanguineum L. with Anti-Influenza Activity
Authors: Pantev Assen , Ivancheva Stefka , Staneva Lidija , Serkedjieva Julia
Published: February 2015

From the aerial roots of the medicinal plant Geranium sanguineum L. a polyphenol-rich extract with strong anti-influenza activity has been isolated. To investigate its active fractions, the extract was partitioned by solvents with increasing polarity. The n-BuOH fraction contained the majority of the in vitro antiviral activity; the EtOAc fraction was the most effective one in vivo. A bioassay-directed fractionation of the n-BuOH and EtOAc fractions was performed to obtain information about the nature of the chemical components of the plant extract, responsible for the antiviral effect. The individual constituents were identified by spectroscopic methods and comparison with authentic samples and by HPLC. The cell-toxic and virus-inhibitory effects of the fractions and some individual polyphenol compounds, found in Geranium sanguineum L., were studied using the replication of representative influenza viruses in cell cultures. This study showed that the presence of a variety of biologically active compounds as well as the possible synergistic interactions between them seem to be decisive for the overall antiviral effect.
Radical Quenching by Rosmarinic Acid from Lavandula vera MM Cell Culture
Authors: Kovacheva Elena , Georgiev Milen , Pashova Svetlana , Ilieva Mladenka
Published: February 2015

This study was conducted to evaluate the radical scavenging capacities of extracts and preparations from a Lavandula vera MM plant cell culture with different rosmarinic acid content and to compare them with pure rosmarinic and caffeic acids as well. The methods, which were used are superoxide anion and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radicals scavenging assays. Results showed that extracts and preparations from Lavandula vera MM possess strong radical scavengers, as the best both radical scavengers appeared to be the fractions with enriched rosmarinic acid content, obtained after ethylacetate fractioning (47.7% inhibition of superoxide radicals and 14.2 μᴍ 6-hydroxy- 2,5,7,8-tetramethylchroman-2-carboxylic acid equivalents, respectively). These data reveal the possibilities for application of these preparations as antioxidants.
Structure-Activity Relationship of Acetylenes from Galls of Hedera rhombea as Plant Growth Inhibitors
Authors: Yamazoe Sayumi , Hasegawa Koji , Shigemori Hideyuki
Published: February 2015

The structure-activity relationship of 12 isolated acetylenes from galls of Hedera rhombea (Araliaceae) induced by Asphondylia sp. (Cecidomyiidae) and their derivatives has been studied for the inhibition of the shoot and root growth of rice, perennial ryegrass, cockscomb, lettuce, and cress. Almost all acetylenes generally showed growth inhibitory activity. The diacetylenes exhibited higher activity than the monoacetylenes, suggesting that a conjugated diyne segment is essential for the activity. On the other hand, the acetylenes with a nonoxidated methylene group at C-8 showed stronger activity comparing with those possessing hydroxy and acetoxy groups at C-8. Furthermore, it has been demonstrated that the acetylenes bearing a terminal olefinic group at C-16,C-17 enhanced the activity. It is thus clarified that important sites for the activity of the acetylenes from galls of H. rhombea are a conjugated diyne and a terminal olefinic group connecting to the aliphatic chain and that less oxidated compounds show more activity.
In vitro Organogenesis and Alkaloid Accumulation in Datura innoxia
Authors: Zayed Rawia , Wink Michael , El-Shamy Hesham
Published: February 2015

The kinetics of tropane alkaloids accumulation in different organs such as roots, leaves, stems, flowers and seeds of Datura innoxia was investigated by GC-MS. Twenty-six tropane alkaloids were detected. The ester derivatives of tropine (3α-tigloyloxytropine and 3-tigloyloxy- 6-hydroxytropine) are the major compounds. Undifferentiated callus were established from the stem explants of Datura innoxia using Murashige and Skoog (MS) medium supplied with 6-benzylaminopurine (BA, 1 mg l-1) and indole-3-acetic acid (IAA, 0.5 mg l-1) in combination for 6 weeks. Callus differentiation was initiated by subculture onto solid MS medium, free from hormones, for more than 10 months. Initially, shoots were formed after four weeks from subculture. Further subculturing in basal MS medium without growth regulators initiated the rooting of a shooty callus after 6 weeks. Investigation of the alkaloid content of the unorganized and organized callus revealed that callus (either green or brown) yielded only trace amounts of alkaloids. On the other hand, re-differentiated shoots contained mainly scopolamine while re-differentiated roots biosynthesized hyoscyamine as the main alkaloid.
Influence of Glyphosate on Flower Morphogenesis and Pigmentation in Petunia hybrida
Authors: Shimada Atsumi , Kimura Yasuo
Published: February 2015

Glyphosate showed a remarkable effect inducing the change of flower symmetry from the actinomorphic to the zygomorphic type in Petunia hybrida. Glyphosate [N-(phosphonomethyl) glycine] reduced the anthocyanin content and showed a weak inhibitory effect against phenylalanine ammonia-lyase (PAL) activity. ʟ-2-Aminooxy-3-phenylpropionic acid (APA), an inhibitor of PAL activity, reduced the anthocyanin content but had no effect on flower shape. Additional phenylalanine or trans-cinnamic acid, the intermediates of glyphosate inhibition against PAL activity, could not recover the change of flower shape induced by glyphosate. These results suggested that the reduction of PAL activity alone could not account for the two characteristic changes of flower symmetry and pigmentation induced by glyphosate. On the other hand, the results of application of glyphosate-related compounds suggested that the structure of glyphosate contributed to induce the morphological change of Petunia flower. Glyphosate may thus be a very useful agent in the elucidation of unresolved questions of flower morphogenesis and the related metabolism.
Overexpression of celB Gene Coding for β-Glucosidase from Pyrococcus furiosus Using a Baculovirus Expression Vector System in Silkworm, Bombyx mori
Authors: Lin Xu’Ai , Zhang Wei , Chen Yin , Yao Bin , Zhang Zhi Fang
Published: February 2015

β-Glucosidase is a member of the glycosyl hydrolases that specifically catalyze the hydrolysis of terminal nonreducing β-ᴅ-glucose residues from the end of various oligosaccharides with the release of β-ᴅ-glucose. CelB gene, encoding the thermostable β-glucosidase, was amplified from the Pyrococcus furiosus genome and then cloned into the baculoviral transfer vector under the control of the polyhedrin gene promoter. After co-transfection with the genetically modified parental Bombyx mori nucleopolyhedrovirus (BmNPV), the recombinant virus containing celB gene was used to express β-glucosidase in silkworm. The recombinant β-glucosidase was purified to about 81% homogeneity in a single heat-treatment step. The optimal activity of the expressed β-glucosidase was obtained at pH 5.0 and about 105 °C; divalent cations and high ionic strength did not affect the activity remarkably. This suggested that the enzymatic characteristics of recombinant β-glucosidase were similar to the native counterpart. The expressed β-glucosidase accounted for more than 10% of silkworm total haemolymph proteins according to the protein quantification and densimeter scanning. The expression level reached 10,199.5 U per ml haemolymph and 19,797.4 U per silkworm larva, and the specific activity of the one-step purified crude enzyme was 885 U per mg. It was demonstrated to be an attractive approach for mass production of thermostable β-glucosidase using this system.
Hydroxamic Acids in Secale cereale L. and the Relationship with their Antifeedant and Allelopathic Properties
Authors: Copaja Sylvia V. , Villarroel Elisa , Bravo Héctor R. , Pizarro Lorena , Argandoñ Víctor H.
Published: February 2015

Contents of the hydroxamic acids 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), and 2,4- dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) in leaves and roots of 14 cultivars of rye, Secale cereale L., were determined. Dynamics of accumulation in three cultivars were evaluated. DIBOA was the main cyclic hydroxamic acid in leaves but the contents differed significantly between the cultivars. Both DIBOA and DIMBOA were present in the roots. Maximum concentration of DIBOA in leaves and DIMBOA in roots was reached between 48-54 h and 54-72 h after germination, respectively. Antifeedant activity of DIBOA towards the aphid Rhopalosiphum padi and the feeding behavior were studied by electronic recording in barley leaves treated with different contents of DIBOA. The deleterious activity of DIBOA could arise by starvation and/or a toxic effect. Additionally, allelopathic potential of pure DIBOA and aqueous extracts of leaves and roots of rye (Tetra-Baer) on the germination of lettuce (Lactuca sativa) and rye (Tetra-Baer) seeds was evaluated. A high percentage of germination inhibition of pure DIBOA and the extracts of leaves and roots was observed. The activity is in agreement with the contents of hydroxamic acids in the plants. The substrates had no allelopathic effect on rye seeds.
Solid State Biosurfactant Production in a Fixed-Bed Column Bioreactor
Authors: Guimarães Martins Vilásia , Kalil Susana Juliano , Elit Telma Elita , Vieira Costa Jorge Alberto
Published: February 2015

Biosurfactants are surface active substances which reduce interfacial tension and are produced or excreted at the microbial cell surface. We evaluated the biosurfactant production by Aspergillus fumigatus and Phialemonium sp. in solid state processes using fixed-bed column reactors. We evaluated two media, rice husks alone (simple support) and rice husks plus defatted rice bran (complex support), both enriched with either soy oil or diesel oil. The highest water-in-oil emulsifying activity (EAw/o) obtained was 7.36 EU g-1 produced by A. fumigatus growing on complex support enriched with soy oil and supplied with air at a rate of 60 mL g-1 h-1, while Phialemonium sp. had a maximum production of 6.11 EU g-1 using the simple support with diesel oil and an aeration rate of 120 mL g-1 h-1. The highest oilin- water emulsifying activity (EAo/w) was 12.21 EU g-1 produced by Phialemonium sp. on the complex support enriched with diesel oil and at an aeration rate of 60 mL g-1 h-1, while A. fumigatus produced a maximum EAo/w of 10.98 EU g-1 when growing on the complex support with no additional carbon source and an aeration rate of 60 mL g-1 h-1.
Non-Invasive Monitoring of the Light-Induced Cyclic Photosynthetic Electron Flow during Cold Hardening in Wheat Leaves
Authors: Apostol Simona , Szalai Gabriella , Sujbert László , Popova Losanka P. , Janda Tibor
Published: February 2015

The effect of irradiance during low temperature hardening was studied in a winter wheat variety. Ten-day-old winter wheat plants were cold-hardened at 5 °C for 11 days under light (250 μmol m-2 s-1) or dark (20 μmol m-2 s-1) conditions. The effectiveness of hardening was significantly lower in the dark, in spite of a slight decrease in the Fv/Fm chlorophyll fluorescence induction parameter, indicating the occurrence of photoinhibition during the hardening period in the light. Hardening in the light caused a downshift in the far-red induced AG (afterglow) thermoluminescence band. The faster dark re-reduction of P700+, monitored by 820-nm absorbance, could also be observed in these plants. These results suggest that the induction of cyclic photosynthetic electron flow may also contribute to the advantage of frost hardening under light conditions in wheat plants.
Antiviral and Antimicrobial Profiles of Selected Isoquinoline Alkaloids from Fumaria and Corydalis Species
Authors: Orhan Ilkay , Özçelik Berrin , Karaoğlu Taner , Şener Bilge
Published: February 2015

In the current study, 33 isoquinoline alkaloids belonging to protopine-, benzylisoquinoline-, benzophenanthridine-, spirobenzylisoquinoline-, phthalideisoquinoline-, aporphine-, protoberberine-, cularine-, and isoquinolone-types as well as 7 derivatives of them obtained from some Fumaria and Corydalis species growing in Turkey have been evaluated for their in vitro antiviral and antimicrobial activities. Both DNA virus Herpes simplex (HSV) and RNA virus Parainfluenza (PI-3) were employed for antiviral assessment of the compounds using Madine- Darby bovine kidney and Vero cell lines and their maximum non-toxic concentrations (MNTC) and cytopathogenic effects (CPE) were determined using acyclovir and oseltamivir as the references. Antibacterial and antifungal activities of the alkaloids were tested against Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Bacillus subtilis, and Candida albicans by the microdilution method and compared to ampicilline, ofloxacine, and ketocanazole as the references. The alkaloids did not present any notable antibacterial effect, while they had significant antifungal activity at 8 μg/ml concentration. On the other hand, the alkaloids were found to have selective inhibition against the PI-3 virus ranging between 0.5 and 64 μg/ml as minimum and maximum CPE inhibitory concentrations, whereas they were completely inactive towards HSV.
Antinociceptive Activity of Structural Analogues of Rotundifolone: Structure-Activity Relationship
Authors: Sousa Damião P. de , Júnior Edison V. M. , Oliveira Fernando S. , Almeida Reinaldo N. de , Nunes Xirley P. , Barbosa-Filho José M.
Published: February 2015

Rotundifolone, a monoterpene isolated from the essential oil of the leaves of Mentha x villosa, is a constituent of several essential oils and known to have antinociceptive activity. Our recent study demonstrated that the analogues of rotundifolone showed also a significant antinociceptive effect. In the present report, to investigate the correlation between the structure and antinociceptive activity, rotundifolone and its analogues were evaluated in the acetic acid-induced writhing test in mice. All compounds showed to be more antinociceptive than rotundifolone against the pain response induced by acetic acid. Comparing the antinociceptive effect of rotundifolone with limonene oxide and (+)-pulegone, the results demonstrated that the epoxide group contributes as much as the ketone group to the antinociceptive activity of rotundifolone. Similarly, pulegone oxide and carvone epoxide were more antinociceptive than rotundifolone, thereby suggesting that the position of the functional group on the ring also influences the antinociceptive activity. (D)-Carvone produced maximal inhibition of the writhing response and was slightly more active than (+)-carvone. The study showed that by appropriate structural modification it may be possible to develop novel antinociceptive agents.
Induction of Phenolic Compounds in Wheat (Triticum aestivum L.) Tissue Cultures by Streptomycin
Authors: Hassan Atiat M. A.
Published: February 2015

The tissue cultures of wheat (Triticum aestivum L.) were induced from the mature embryos (explants) of the dry grains and grown on MS medium containing kinetin (0.1 mg/l) and 2,4 D (1.0 mg/l). The cultures were incubated for two weeks at (25 ± 2) °C under a light/dark regime (16 h light daily). The formed calli were subcultured at the beginning of the stationary growth phase (15 days) with fresh MS medium containing 0, 5, 10, 25, 50, 100, 150 mg/l streptomycin elicitor and maintained for two weeks for three subcultures. A significant increase in phenylalanine ammonia lyase (PAL) activity coincided with the increase of the total phenolic compounds after elicitation with streptomycin. Maximum induction was recorded during the first two weeks, then gradually declined during the rest of the experimental period, but the values attained were still markedly higher than that of the control. The endogenous cinnamic acid content was also increased significantly with the increase in PAL activity making about 2-18% of the total phenolic acids. The growth and accumulation of phenolic compounds were inversely related. However, accumulation of phenolic compounds became limited for growth of wheat tissue culture especially during the long term cultivation.
Ultracytochemical Localization of Acid Phosphatase in Humicola lutea Conidia and Mycelia
Authors: Spasova Dimitrina , Aleksieva Penka , Nacheva Lilyana , Radoevska Spasimira
Published: February 2015

Electron microscopic cytochemical procedures were used to determine the cellular location of acid phosphatase in the fungus Humicola lutea grown in casein-containing medium lacking in mineral orthophosphates. In our investigations acid phosphatase in nongerminating conidia was localized on the outer side of the cell wall, in the cell wall, and on the exterior surface of the plasma membrane. The reaction product of acid phosphatase in germinating conidia was seen in the outer wall layer while in young mycelium on the cell surface and in the exocellular space. The relationship between phosphatase activities localized in the cell wall and their role in the enzymatic degradation of the phosphoprotein casein providing available phosphates for cell growth is discussed.
Effect of Copper on Acid Phosphatase Activity in Yeast Yarrowia lipolytica
Authors: Ito Hiroyasu , Inouhe Masahiro , Tohoyama Hiroshi , Joho Masanori
Published: February 2015

Acid phosphatase (APase) activity of the yeast Yarrowia lipolytica increased with increasing Cu2+ concentrations in the medium. Furthermore, the enzyme in soluble form was stimulated in vitro by Cu2+, Co2+, Ni2+, Mn2+ and Mg2+ and inhibited by Ag+ and Cd2+. The most effective ion was Cu2+, especially for the enzyme from cultures in medium containing Cu2+, whereas APase activity in wall-bound fragments was only slightly activated by Cu2+. The content of cellular phosphate involving polyphosphate was decreased by adding Cu2+, regardless of whether or not the medium was rich in inorganic phosphate. Overproduction of the enzyme stimulated by Cu2+ might depend on derepression of the gene encoding the APase isozyme.
Nuclear-Localized Plastid DNA Fragments in Protozoa, Metazoa and Fungi
Authors: Yuan Shu , Sun Xin , Mu Lin-Chun , Liu Wen-Juan , Lei Tao , Wang Jian-Hui , Du Jun-Bo , Lin Hong-Hui
Published: February 2015

We analyzed nuclear-localized plastid-like DNA (nupDNA) fragments in protozoa, metazoa and fungi. Most eukaryotes that do not have plastids contain 40-5000 bp nupDNAs in their nuclear genomes. These nupDNA fragments are mainly derived from repeated regions of plastids and distribute through the whole genomes. A majority of nupDNA fragments is located on coding regions with very important functions. Similar to plastids, these nupDNAs most possibly originate from cyanobacteria. Analysis of them suggests that through millions of years of universal endosymbiosis and gene transfer they may have occurred in ancient protists before divergence of plants and animals/fungi, and some transferred fragments have been reserved till now even in modern mammals.
New Findings about Ellman’s Method to Determine Cholinesterase Activity
Authors: Komersová Alena , Komers Karel , Čegan Alexandr
Published: February 2015

The original Ellman’s spectrophotometrical method for cholinesterase activity determination uses 5,5′-dithiobis-2-nitrobenzoic acid (DTNB, Ellman’s reagent) as a chromogen and records the level of cholinesterase activity as an increase of absorbance at 412 nm. Although this procedure usually poses no problem, exceptions arise when the concentration of DTNB is far higher than the concentration of acetylthiocholine (ATCH). It was found that the ratio of concentrations of DTNB/ATCH is an important parameter for the ATCH hydrolysis course: high excess of DTNB decreases the hydrolysis rate resulting in a lower measured enzyme activity. Our experiments indicate that this influence of DTNB concentration can be explained by the inhibition of ATCH hydrolysis by DTNB.
Pigments of Fly Agaric (Amanita muscaria)
Authors: Stintzing Florian , Schliemann Willibald
Published: February 2015

The complex pigment pattern of fly agaric (Amanita muscaria) cap skins has been studied by LC-DAD and mass spectrometry. Among the betaxanthins the corresponding derivatives of serine, threonine, ethanolamine, alanine, Dopa, phenylalanine and tryptophan are reported for the first time to contribute to the pigment pattern of fly agarics. Betalamic acid, the chromophoric precursor of betaxanthins and betacyanins, muscaflavin and seco-dopas were also detected. Furthermore, the red-purple muscapurpurin and the red muscarubrin were tentatively assigned while further six betacyanin-like components could not be structurally allocated. Stability studies indicated a high susceptibility of pigment extracts to degradation which led to rapid colour loss thus rendering a complete characterization of betacyaninlike compounds impossible at present. Taking into account these difficulties the presented results may be a starting point for a comprehensive characterization of the pigment composition of fly agarics.
Bioactive Compounds of the Volatile Oil of Dracocephalum kotschyi
Authors: Saeidnia Soodabeh , Gohari Ahmad Reza , Hadjiakhoondi Abbas , Shafiee Abbas
Published: February 2015

Trypanocidal activity was found in the volatile oil of dried Dracocephalum kotschyi. GCMS analysis determined that the major constituents of the oil were geranial (35.8%), C10H14O (26.6%), limonene (15.8%) and 1,1-dimethoxy decane (14.5%). In order to isolate the unknown biologically active monoterpene, fractionation of the volatile oil was carried out by silica gel column chromatography. The structure of the oxygenated compound was confirmed to be limonene-10-al (C10H14O) by analysis of physical and spectroscopic data (1H NMR, 13C NMR, HMBC and HMQC).