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Sample records for clover hydroxycinnamoyl transferase

  1. Co-downregulation of the hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyl transferase and coumarate 3-hydroxylase significantly increases cellulose content in transgenic alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Tong, Zongyong; Li, Heng; Zhang, Rongxue; Ma, Lei; Dong, Jiangli; Wang, Tao

    2015-10-01

    Lignin is a component of the cell wall that is essential for growth, development, structure and pathogen resistance in plants, but high lignin is an obstacle to the conversion of cellulose to ethanol for biofuel. Genetically modifying lignin and cellulose contents can be a good approach to overcoming that obstacle. Alfalfa (Medicago sativa L.) is rich in lignocellulose biomass and used as a model plant for the genetic modification of lignin in this study. Two key enzymes in the lignin biosynthesis pathway-hydroxycinnamoyl -CoA:shikimate hydroxycinnamoyl transferase (HCT) and coumarate 3-hydroxylase (C3H)-were co-downregulated. Compared to wild-type plants, the lignin content in the modified strain was reduced by 38%, cellulose was increased by 86.1%, enzyme saccharification efficiency was increased by 10.9%, and cell wall digestibility was increased by 13.0%. The modified alfalfa exhibited a dwarf phenotype, but normal above ground biomass. This approach provides a new strategy for reducing lignin and increasing cellulose contents and creates a new genetically modified crop with enhanced value for biofuel. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  2. Engineering of Escherichia coli for the synthesis of N-hydroxycinnamoyl tryptamine and serotonin.

    Science.gov (United States)

    Lee, Su Jin; Sim, Geun-Young; Lee, Youngshim; Kim, Bong-Gyu; Ahn, Joong-Hoon

    2017-11-01

    Plants synthesize various phenol amides. Among them, hydroxycinnamoyl (HC) tryptamines and serotonins exhibit antioxidant, anti-inflammatory, and anti-atherogenic activities. We synthesized HC-tryptamines and HC-serotonin from several HCs and either tryptamine or serotonin using Escherichia coli harboring the 4CL (4-coumaroyl CoA ligase) and CaHCTT [hydroxycinnamoyl-coenzyme A:serotonin N-(hydroxycinnamoyl)transferase] genes. E. coli was engineered to synthesize N-cinnamoyl tryptamine from glucose. TDC (tryptophan decarboxylase) and PAL (phenylalanine ammonia lyase) along with 4CL and CaHCTT were introduced into E. coli and the phenylalanine biosynthetic pathway of E. coli was engineered. Using this strategy, approximately 110.6 mg/L of N-cinnamoyl tryptamine was synthesized. By feeding 100 μM serotonin into the E. coli culture, which could induce the synthesis of cinnamic acid or p-coumaric acid, more than 99 μM of N-cinnamoyl serotonin and N-(p-coumaroyl) serotonin were synthesized.

  3. Clover, Red (Trifolium pretense)

    Science.gov (United States)

    Genetic modification of plants by the insertion of transgenes can be a powerful experimental approach to answer basic questions about gene product function. This technology can also be used to make improved crop varieties for use in the field. To apply this powerful tool to red clover, an important ...

  4. Biosynthesis of hydroxycinnamoyl esters and amides in legume species

    Science.gov (United States)

    In forage crops, protein that is degraded following harvest is poorly utilized by ruminant animals, resulting in both economic and environmental consequences. In red clover, secondary reactions of quinones resulting from polyphenol oxidase (PPO)-mediated oxidation of the caffeic acid derivatives pha...

  5. The Clover detector

    Energy Technology Data Exchange (ETDEWEB)

    Beck, F A; Byrski, Th; Durien, D; Duchene, G; France, G de; Kharraja, B; Wei, L [Strasbourg-1 Univ., 67 (France). Centre de Recherches Nucleaires; Butler, P; Jones, G; Jones, P [Liverpool Univ. (United Kingdom). Oliver Lodge Lab.; Hannachi, F [Daresbury Lab. (United Kingdom)

    1992-08-01

    The EUROGAM Phase I device is almost running for experiments and new technical developments are in progress for its second phase. For example, a composite Ge detector should enable: a very large photopeak efficiency with good energy and timing resolutions; and, the covering, with Ge, of a large portion of 4{pi}-Str. The Clover detector, proposed by the CRN, Strasbourg, is one of this new generation of Ge detectors. It is currently developed in France by the EUROGAM collaboration. The design, the technical characteristics of the counter and the first results of the prototype tests are discussed in this contribution. (author). 1 ref., 2 tabs., 2 refs.

  6. Lignification in transgenics deficient in 4-coumarate 3-hydroxylase (C3H)or the associated hydroxycinnamoyl transferase (HCT)

    Science.gov (United States)

    John Ralph; Takuya Akiyama; Hoon Kim; Fachuang Lu; Sally A. Ralph; Clint Chapple; Ramesh B. Nair; Armin Wagner; Fang Chen; M.S. Srinivasa Reddy; Richard A Dixon; Heather D. Coleman; Shawn D. Mansfield

    2006-01-01

    Down-regulation of the gene encoding 4-coumarate 3-hydroxylase (C3H) in angiosperms massively but predictably increased the proportion of p-hydroxyphenyl (P) units relative to the normally dominant syringyl (S) and guaiacyl (G) units. Alfalfa stem levels of up to ~65% P (from wild-type (WT) levels of ~1%) resulting from down-regulation of C3H were measured by...

  7. Transferases in Polymer Chemistry

    NARCIS (Netherlands)

    van der Vlist, Jeroen; Loos, Katja; Palmans, ARA; Heise, A

    2010-01-01

    Transferases are enzymes that catalyze reactions in which a group is transferred from one compound to another. This makes these enzymes ideal catalysts for polymerization reactions. In nature, transferases are responsible for the synthesis of many important natural macromolecules. In synthetic

  8. Fold distributions at clover, crystal and segment levels for segmented clover detectors

    International Nuclear Information System (INIS)

    Kshetri, R; Bhattacharya, P

    2014-01-01

    Fold distributions at clover, crystal and segment levels have been extracted for an array of segmented clover detectors for various gamma energies. A simple analysis of the results based on a model independant approach has been presented. For the first time, the clover fold distribution of an array and associated array addback factor have been extracted. We have calculated the percentages of the number of crystals and segments that fire for a full energy peak event

  9. Measurements on a prototype segmented Clover detector

    CERN Document Server

    Shepherd, S L; Cullen, D M; Appelbe, D E; Simpson, J; Gerl, J; Kaspar, M; Kleinböhl, A; Peter, I; Rejmund, M; Schaffner, H; Schlegel, C; France, G D

    1999-01-01

    The performance of a segmented Clover germanium detector has been measured. The segmented Clover detector is a composite germanium detector, consisting of four individual germanium crystals in the configuration of a four-leaf Clover, housed in a single cryostat. Each crystal is electrically segmented on its outer surface into four quadrants, with separate energy read-outs from nine crystal zones. Signals are also taken from the inner contact of each crystal. This effectively produces a detector with 16 active elements. One of the purposes of this segmentation is to improve the overall spectral resolution when detecting gamma radiation emitted following a nuclear reaction, by minimising Doppler broadening caused by the opening angle subtended by each detector element. Results of the tests with sources and in beam will be presented. The improved granularity of the detector also leads to an improved isolated hit probability compared with an unsegmented Clover detector. (author)

  10. Modeling of clover detector in addback mode

    Science.gov (United States)

    Kshetri, R.

    2012-07-01

    Based on absorption and scattering of gamma-rays, a formalism has been presented for modeling the clover germanium detector in addback mode and to predict its response for high energy γ-rays. In the present formalism, the operation of a bare clover detector could be described in terms of three quantities only. Considering an additional parameter, the formalism could be extended for suppressed clover. Using experimental data on relative single crystal efficiency and addback factor as input, the peak-to-total ratio has been calculated for three energies (Eγ = 3.401, 5.324 and 10.430 MeV) where direct measurement of peak-to-total ratio is impossible due to absence of a radioactive source having single monoenergetic gamma-ray of that energy. The experimental validation and consistency of the formalism have been shown considering data for TIGRESS clover detector. In a recent work (R. Kshetri, JINST 2012 7 P04008), we showed that for a given γ-ray energy, the formalism could be used to predict the peak-to-total ratio as a function of number of detector modules. In the present paper, we have shown that for a given composite detector (clover detector is considered here), the formalism could be used to predict the peak-to-total ratio as a function of γ-ray energy.

  11. A first principle approach for clover detector

    Science.gov (United States)

    Kshetri, R.

    2012-08-01

    A simple model based on probability flow arguments has been presented for understanding the clover germanium detector. Using basic concepts of absorption and scattering of gamma-rays, the operation of the clover detector has been described in terms of six probability amplitudes and a parameter. Instead of using an empirical method or simulation, this work presents the first attempt to calculate the peak-to-total and peak-to-background ratios of the clover detector using experimental data of relative single crystal efficiency and addback factor as an input. A unique feature of our approach is that these ratios could be calculated for energies where their direct measurement is impossible due to absence of a radioactive source having single monoenergetic gamma-ray of that energy. Results for four gamma-ray energies (Eγ = 1.408, 3.907, 7.029 and 10.430 MeV) have been discussed. Agreement between experimental data and analysis results has been observed. The present approach could describe clover-type detectors as well. As an example, the nine element detector has been considered. We have demonstrated that our formalism can describe both finite and infinite interactions of γ-rays with the clover crystals. The work presented in this paper follows similar philosophy as presented in a recent paper (R. Kshetri, JInst 2012 7 P04008), which deals with modeling of encapsulated type composite detectors like miniball, cluster and SPI (Spectrometer for INTEGRAL satellite).

  12. Modeling of clover detector in addback mode

    International Nuclear Information System (INIS)

    Kshetri, R

    2012-01-01

    Based on absorption and scattering of gamma-rays, a formalism has been presented for modeling the clover germanium detector in addback mode and to predict its response for high energy γ-rays. In the present formalism, the operation of a bare clover detector could be described in terms of three quantities only. Considering an additional parameter, the formalism could be extended for suppressed clover. Using experimental data on relative single crystal efficiency and addback factor as input, the peak-to-total ratio has been calculated for three energies (E γ = 3.401, 5.324 and 10.430 MeV) where direct measurement of peak-to-total ratio is impossible due to absence of a radioactive source having single monoenergetic gamma-ray of that energy. The experimental validation and consistency of the formalism have been shown considering data for TIGRESS clover detector. In a recent work (R. Kshetri, JINST 2012 7 P04008), we showed that for a given γ-ray energy, the formalism could be used to predict the peak-to-total ratio as a function of number of detector modules. In the present paper, we have shown that for a given composite detector (clover detector is considered here), the formalism could be used to predict the peak-to-total ratio as a function of γ-ray energy.

  13. A first principle approach for clover detector

    International Nuclear Information System (INIS)

    Kshetri, R

    2012-01-01

    A simple model based on probability flow arguments has been presented for understanding the clover germanium detector. Using basic concepts of absorption and scattering of gamma-rays, the operation of the clover detector has been described in terms of six probability amplitudes and a parameter. Instead of using an empirical method or simulation, this work presents the first attempt to calculate the peak-to-total and peak-to-background ratios of the clover detector using experimental data of relative single crystal efficiency and addback factor as an input. A unique feature of our approach is that these ratios could be calculated for energies where their direct measurement is impossible due to absence of a radioactive source having single monoenergetic gamma-ray of that energy. Results for four gamma-ray energies (E γ = 1.408, 3.907, 7.029 and 10.430 MeV) have been discussed. Agreement between experimental data and analysis results has been observed. The present approach could describe clover-type detectors as well. As an example, the nine element detector has been considered. We have demonstrated that our formalism can describe both finite and infinite interactions of γ-rays with the clover crystals. The work presented in this paper follows similar philosophy as presented in a recent paper (R. Kshetri, JInst 2012 7 P04008), which deals with modeling of encapsulated type composite detectors like miniball, cluster and SPI (Spectrometer for INTEGRAL satellite).

  14. Crystal Structure of Green Fluorescent Protein Clover and Design of Clover-Based Redox Sensors.

    Science.gov (United States)

    Campbell, Benjamin C; Petsko, Gregory A; Liu, Ce Feng

    2018-02-06

    We have determined the crystal structure of Clover, one of the brightest fluorescent proteins, and found that its T203H/S65G mutations relative to wild-type GFP lock the critical E222 side chain in a fixed configuration that mimics the major conformer of that in EGFP. The resulting equilibrium shift to the predominantly deprotonated chromophore increases the extinction coefficient (EC), opposes photoactivation, and is responsible for the bathochromic shift. Clover's brightness can further be attributed to a π-π stacking interaction between H203 and the chromophore. Consistent with these observations, the Clover G65S mutant reversed the equilibrium shift, dramatically decreased the EC, and made Clover photoactivatable under conditions that activated photoactivatable GFP. Using the Clover structure, we rationally engineered a non-photoactivatable redox sensor, roClover1, and determined its structure as well as that of its parental template, roClover0.1. These high-resolution structures provide deeper insights into structure-function relationships in GFPs and may aid the development of excitation-improved ratiometric biosensors. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Modelling interactions in grass-clover mixtures

    NARCIS (Netherlands)

    Nassiri Mahallati, M.

    1998-01-01

    The study described in this thesis focuses on a quantitative understanding of the complex interactions in binary mixtures of perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) under cutting. The first part of the study describes the dynamics of growth, production

  16. Application of various technological processes in red clover seed processing

    OpenAIRE

    Đokić, Dragoslav; Stanisavljević, Rade; Terzić, Dragan; Marković, Jordan; Radivojević, Gordana; Anđelković, Bojan; Barać, Saša

    2012-01-01

    This paper presents the results of the processing of natural red clover seed on the processing equipment using different technological methods. Red clover seed, for the establishment and crop utilization, must be of high purity, germination, and high genetic values. These requirements are achieved by processing, or removing impurities and poor quality seeds. Red clover seed processing involves a number of operations, of which the most important are: cleaning, packaging, labeling and storage. ...

  17. White clover regenerative ability under N fertilizing and grazing

    Directory of Open Access Journals (Sweden)

    Josip Leto

    2008-12-01

    Full Text Available Recently, ecological and economic factors in milk and meat production stimulate use of legumes and grass-legumes mixtures, with zero or minimum mineral N as alternative to grass monoculture with high rate of mineral N. Research objective was to examine the effect of N application (0-N0 and 150 kg ha-1 year-1-N150 and rotational grazing by cattle (C and sheep (S on white clover: growing points number, stolon lenght, stolon dry weight, dry matter yield and clover contribution to total annual herbage production. N150 significantly reduced the growing points number, stolon length and stolon dry weight for more than 70 % compared to N0. Grazing treatment affected stolon population density only in interaction with N application because of N150 significantly reduced white clover population density only in sheep grazing. S-treatment had higher clover DM yield (0.21 t ha-1 than C-treatment (0.13 t ha-1. N0 had higher clover DM yield (0.25 t ha-1 than N150 (0.09 t ha-1. However, the interaction grazing management x N rate was significant for clover DM yield and clover contribution to total DM yield. N150 reduced both parameters for 80 % only in sheep grazing while difference in DM yield and clover contribution to total DM yield between grazing treatment was recorded only in N0 Sheep grazing increased DM yield for 150 % and clover contribution for 99 % compared to cattle grazing.

  18. Complementary effects of red clover inclusion in ryegrass-white clover swards for grazing and cutting

    DEFF Research Database (Denmark)

    Eriksen, Jørgen; Askegaard, Margrethe; Søegaard, Karen

    2014-01-01

    Increasing plant species diversity in grasslands may improve productivity and stability of yields. In a field experiment, we investigated the herbage dry-matter (DM) yield and crude protein content of two-species swards of perennial ryegrass–white clover (Lolium perenne L.–Trifolium repens L...... clover in sown swards are discussed. These may include higher nitrogen-use efficiency in ruminants, increased soil fertility and improved sward flexibility to cope with changing managements. The findings also suggest positive yield effects of alternating between cutting and grazing within the season...

  19. Defective Pollen Wall 2 ( DPW2 ) Encodes an Acyl Transferase Required for Rice Pollen Development

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Dawei [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; Shi, Jianxin [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; Rautengarten, Carsten [Univ. of Melbourne (Australia). ARC Centre of Excellence in Plant Cell Walls; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Inst. and Biological Systems and Engineering Division; Yang, Li [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; Qian, Xiaoling [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; Uzair, Muhammad [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; Zhu, Lu [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; Luo, Qian [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; An, Gynheung [Kyung Hee Univ., Yongin (Korea). Crop Biotech Inst.; Waßmann, Fritz [Univ. of Bonn (Germany). Inst. of Cellular and Molecular Botany; Schreiber, Lukas [Univ. of Bonn (Germany). Inst. of Cellular and Molecular Botany; Heazlewood, Joshua L. [Univ. of Melbourne (Australia). ARC Centre of Excellence in Plant Cell Walls; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Inst. and Biological Systems and Engineering Division; Scheller, Henrik Vibe [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Inst. and Biological Systems and Engineering Division; Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology; Hu, Jianping [Michigan State Univ., East Lansing, MI (United States). Dept. of Energy Plant Research Lab.; Zhang, Dabing [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences; Univ. of Adelaide, SA (Australia). School of Agriculture, Food and Wine; Liang, Wanqi [Shanghai Jiao Tong Univ. (China). Joint International Research Lab. of Metabolic and Developmental Sciences

    2016-05-31

    Aliphatic and aromatic lipids are both essential structural components of the plant cuticle, an important interface between the plant and environment. Although cross links between aromatic and aliphatic or other moieties are known to be associated with the formation of leaf cutin and root and seed suberin, the contribution of aromatic lipids to the biosynthesis of anther cuticles and pollen walls remains elusive. In this study, we characterized the rice (Oryza sativa) male sterile mutant, defective pollen wall 2 (dpw2), which showed an abnormal anther cuticle, a defective pollen wall, and complete male sterility. Compared with the wild type, dpw2 anthers have increased amounts of cutin and waxes and decreased levels of lipidic and phenolic compounds. DPW2 encodes a cytoplasmically localized BAHD acyltransferase. In vitro assays demonstrated that recombinant DPW2 specifically transfers hydroxycinnamic acid moieties, using v-hydroxy fatty acids as acyl acceptors and hydroxycinnamoyl-CoAs as acyl donors. Thus, The cytoplasmic hydroxycinnamoyl-CoA:v-hydroxy fatty acid transferase DPW2 plays a fundamental role in male reproduction via the biosynthesis of key components of the anther cuticle and pollen wall.

  20. Cytoskeleton-amyloplast interactions in sweet clover

    Science.gov (United States)

    Guikema, J. A.; Hilaire, E.; Odom, W. R.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    The distribution of organelles within columella cells of sweet clover was examined by transmission electron microscopy following growth under static or clinorotating conditions. A developmentally conditioned polarity was observed, with a proximal location of the nucleus and a distal accumulation of the endoplasmic reticulum. This polarity was insensitive to clinorotation. In contrast, clinorotation altered the location of amyloplasts. Application of cytoskeletal poisons (colchicine, cytochalasin D, taxol, and phalloidin), especially during clinorotation, had interesting effects on the maintenance of columella cell polarity, with a profound effect on the extent, location, and structure of the endoplasmic reticulum. The site of cytoskeletal interactions with sedimenting amyloplasts is thought to be the amyloplast envelope. An envelope fraction, having over 17 polypeptides, was isolated using immobilized antibody technology, and will provide a means of assessing the role of specific peptides in cytoskeleton/amyloplast interactions.

  1. Carnitine palmityl transferase I deficiency

    NARCIS (Netherlands)

    Al-Aqeel, A. I.; Rashed, M. S.; Ruiter, J. P.; Al-Husseini, H. F.; Al-Amoudi, M. S.; Wanders, R. J.

    2001-01-01

    Carnitine palmityl transferase I is the key enzyme in the carnitine dependent transport of long chain fatty acids across the mitochondrial inner membrane and its deficiency results in a decrease rate of fatty acids beta-oxidation with decreased energy production. We reported a family of 3 affected

  2. Inhibition of hydroxycinnamoyl-CoA thioesterases in ginger (Zingiber officinale Rosc.) and turmeric (Curcuma longa L.) by lipase inhibitors.

    Science.gov (United States)

    Flores-Sanchez, Isvett Josefina; Gang, David Roger

    2013-11-01

    Ginger (Zingiber officinale Rosc.) and turmeric (Curcuma longa L.), members of the Zingiberaceae, are widely used in traditional Asian cuisines and herbal medicine. Gingerols and diarylheptanoids, important compounds from these plants, appear to be produced by enzymes of the type III polyketide synthase class. Previous efforts to detect activity of such enzymes in tissues from these plants were only marginally successful in turmeric and completely unsuccessful in ginger because of very rapid hydrolysis of the hydroxycinnamoyl-CoA substrates (p-coumaroyl-CoA, feruloyl-CoA and caffeoyl-CoA) in these assays, presumably due to the presence of thioesterases in these tissues. In order to determine whether such thioesterase activities were specific and could be reduced so that the polyketide synthase activities could be better characterized, three inhibitors of the thioesterase domain of fatty acid synthase were tested in assays with leaf and rhizome crude protein extracts from these plants: orlistat, a reduced form of lipstatin, and peptide 1 and peptide 2 from hydrolysates of soybean β-conglycinin. Results of these analyses indicated that specific thioesterases do exist in these plants and that they could indeed be inhibited, with highest inhibition occurring with a mixture of these three compounds, leading for example to a reduction of caffeoyl-CoA hydrolysis in leaves and rhizomes of ginger by 40-fold and 27-fold, respectively. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  3. Biosynthesis of phenolic compounds inVitis vinifera cell suspension cultures: Study on hydroxycinnamoyl CoA:ligase.

    Science.gov (United States)

    Lotfy, S; Lofty, S; Fleuriet, A; Ramos, T; Macheix, J J

    1989-02-01

    In cell suspensions cultures from grape berry pulp (Vitis vinifera cv. Gamay fréaux)hydroxycinnamoyl CoA ligase (CoAL) displayed maximum activity (100 %) forp-coumaric acid and then, in decreasing order, for ferulic acid (81.3 %) and caffeic acid (60.4 %). No activity was detected with sinapic and cinnamic acids. The changes in CoAL activity during the growth cycle of the culture displayed two peaks : the highest (6 h after subculturing) was linked with a strong increase in protein caused by dilution ; the second was weaker and occurred on the 7th day of culture.Grape cell suspension accumulated mainly peonidin (Pn) and cyanidin (Cy) glucosides (Pn 3-glucoside, Cy 3-glucoside, Pn 3-acetylglucoside, Pn 3-caffeylglucoside, Pn 3-p-coumarylglucoside, and Cy 3-p-coumarylglucoside). Maximum accumulation of anthocyanins was associated with the exponential growth phase of the culture and might be the result of the substantial increase in CoAL activity resulting from the effect of dilution. The second enzyme activity peak was probably oriented towards the acylation of anthocyanins since the percentage of acylated forms increased with time after subculturing.

  4. Predicting Dry Matter Composition of Clover Grass Leys Using Data Simulation and Camera-based Segmentation of Field Canopies into White Clover, Red Clover, Grass and Weeds

    DEFF Research Database (Denmark)

    Skovsen, Søren; Dyrmann, Mads; Eriksen, Jørgen

    2018-01-01

    species in the biomass. In our setup, we exploit the top-down canopy view of the clover grass ley to estimate the volumetric composition of the yield, and predict the composition of the dry matter of the forage. Using a deep learning approach, the canopy image is automatically pixel-wise segmented....... The biggest hindrance to training a fully convolutional deep neural network is the requirement of labeled data. Due to the complexity, the high number of leaves and high levels of occlusions in clover grass canopies, hand labeling the data requires roughly 20 hours of manual labor per image. The need...... for hundreds or thousands labeled training images renders this approach unfeasible. We have shown that implementation of image simulation of distinct clover grass fields can reduce the labeling task significantly. Investing less than 20 hours of labor, thousands of simulated images and corresponding labels can...

  5. Identification and cloning of an NADPH-dependent hydroxycinnamoyl-CoA double bond reductase involved in dihydrochalcone formation in Malus×domestica Borkh.

    Science.gov (United States)

    Ibdah, Mwafaq; Berim, Anna; Martens, Stefan; Valderrama, Andrea Lorena Herrera; Palmieri, Luisa; Lewinsohn, Efraim; Gang, David R

    2014-11-01

    The apple tree (Malus sp.) is an agriculturally and economically important source of food and beverages. Many of the health beneficial properties of apples are due to (poly)phenolic metabolites that they contain, including various dihydrochalcones. Although many of the genes and enzymes involved in polyphenol biosynthesis are known in many plant species, the specific reactions that lead to the biosynthesis of the dihydrochalcone precursor, p-dihydrocoumaroyl-CoA (3), are unknown. To identify genes involved in the synthesis of these metabolites, existing genome databases of the Rosaceae were screened for apple genes with significant sequence similarity to Arabidopsis alkenal double bond reductases. Herein described are the isolation and characterization of a Malus hydroxycinnamoyl-CoA double bond reductase, which catalyzed the NADPH-dependent reduction of p-coumaroyl-CoA and feruloyl-CoA to p-dihydrocoumaroyl-CoA and dihydroferuloyl-CoA, respectively. Its apparent Km values for p-coumaroyl-CoA, feruloyl-CoA and NADPH were 96.6, 92.9 and 101.3μM, respectively. The Malus double bond reductase preferred feruloyl-CoA to p-coumaroyl-CoA as a substrate by a factor of 2.1 when comparing catalytic efficiencies in vitro. Expression analysis of the hydroxycinnamoyl-CoA double bond reductase gene revealed that its transcript levels showed significant variation in tissues of different developmental stages, but was expressed when expected for involvement in dihydrochalcone formation. Thus, the hydroxycinnamoyl-CoA double bond reductase appears to be responsible for the reduction of the α,β-unsaturated double bond of p-coumaroyl-CoA, the first step of dihydrochalcone biosynthesis in apple tissues, and may be involved in the production of these compounds. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Preliminary Results of Clover and Grass Coverage and Total Dry Matter Estimation in Clover-Grass Crops Using Image Analysis

    Directory of Open Access Journals (Sweden)

    Anders K. Mortensen

    2017-12-01

    Full Text Available The clover-grass ratio is an important factor in composing feed ratios for livestock. Cameras in the field allow the user to estimate the clover-grass ratio using image analysis; however, current methods assume the total dry matter is known. This paper presents the preliminary results of an image analysis method for non-destructively estimating the total dry matter of clover-grass. The presented method includes three steps: (1 classification of image illumination using a histogram of the difference in excess green and excess red; (2 segmentation of clover and grass using edge detection and morphology; and (3 estimation of total dry matter using grass coverage derived from the segmentation and climate parameters. The method was developed and evaluated on images captured in a clover-grass plot experiment during the spring growing season. The preliminary results are promising and show a high correlation between the image-based total dry matter estimate and the harvested dry matter ( R 2 = 0.93 with an RMSE of 210 kg ha − 1 .

  7. Impact on Clover-Grass Yield from Wheel Load and Tyre Pressure

    DEFF Research Database (Denmark)

    Green, Ole; Jørgensen, Rasmus Nyholm; Sørensen, Claus Aage Grøn

    2009-01-01

    Traffic intensities have been shown to have a negative influence on the yield of grass and clover. A full scale grass-clover field trial was established to estimate the effect on clover-grass yields as a function of different wheel loads and tire pressures. The trial comprised 16 different traffi...

  8. Potential for yield in red clover (Trifolium pratense L. varieties

    Directory of Open Access Journals (Sweden)

    Vasiljević Sanja

    2010-01-01

    Full Text Available Red clover (Trifolium pratense L. is botanically a perennial, although it often behaves as a biennial. The crop usually lives 2 to 3 seasons in most clover-growing regions of the world and is generally conceded to be lacking in persistence. The objective of this investigation was to analyze the production potential and persistence of domestic and imported red clover varieties. Experiments were conducted at Rimski Šančevi Experiment Field during the period 2004-2006. The experimental materials were six varieties (K-17, Kolubara, Una, Viola, Start, Nike which were analyzed for the most important production characteristics: yield of green mass, dry matter yield and persistence. .

  9. Estimating the content of clover and grass in the sward using a consumer camera and image processing

    DEFF Research Database (Denmark)

    Mortensen, Anders Krogh; Karstoft, Henrik; Søegaard, Karen

    the dry matter ratio of clover and grass in clover grass fields from sparse close up images. First, the light conditions is determined, which is used for selecting model parameters to estimate the coverage of both clover and grass. Next, the clover and grass coverage are transformed to give the dry matter...

  10. Hibiscus cannabinus feruloyl-coa:monolignol transferase

    Science.gov (United States)

    Wilkerson, Curtis; Ralph, John; Withers, Saunia; Mansfield, Shawn D.

    2016-11-15

    The invention relates to isolated nucleic acids encoding a feruloyl-CoA:monolignol transferase and feruloyl-CoA:monolignol transferase enzymes. The isolated nucleic acids and/or the enzymes enable incorporation of monolignol ferulates into the lignin of plants, where such monolignol ferulates include, for example, p-coumaryl ferulate, coniferyl ferulate, and/or sinapyl ferulate. The invention also includes methods and plants that include nucleic acids encoding a feruloyl-CoA:monolignol transferase enzyme and/or feruloyl-CoA:monolignol transferase enzymes.

  11. Association of phytoplasmas and viruses with malformed clovers

    Czech Academy of Sciences Publication Activity Database

    Fránová, Jana; Paltrinieri, S.; Botti, S.; Šimková, Marie; Bertaccini, A.

    2004-01-01

    Roč. 49, č. 5 (2004), s. 617-624 ISSN 0015-5632 R&D Projects: GA AV ČR IBS5051014 Institutional research plan: CEZ:AV0Z5051902 Keywords : mycoplasma-like organismus * viruses * clovers * classification Subject RIV: EE - Microbiology, Virology Impact factor: 1.034, year: 2004

  12. Genetic variation of white clover ( Trifolium repens L.) collections ...

    African Journals Online (AJOL)

    amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) to investigate the genetic relationships among white clover germplasms of China, and four commercial cultivars were included for a comparison. The results revealed that the populations showed diverse morphological traits, RAPD and SSR patterns.

  13. Genetic variation of white clover (Trifolium repens L.) collections ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-05-24

    May 24, 2010 ... incompatibility mechanism of disomic inheritance (Thomas,. 1987). In essence ..... Badr A, Sayed-Ahmed H, El-Shanshouri A, Watson LE (2002). Ancestors of white ... Caradus JR, Woodfield DR (1997). World checklist of ... within a natural population of white clover (Trifolium repense L). J. Ecol. 73: 615-624 ...

  14. Growth of bermudagrass with white clover or nitrogen fertilizer

    Science.gov (United States)

    White clover (Trifolium repens) var ‘Durana’ was oversown into established bermudagrass (Cynodon dactylon) in 2009. Soil analysis indicated potassium (K) was low and potash at 112 and 336 kg/ha was added as main plots. Nitrogen as ammonium nitrate or an ammonium sulfate/urea blend was added as 0, 34...

  15. Salt tolerance in red clover (Trifolium pratense L.) seedlings

    African Journals Online (AJOL)

    Jane

    2011-08-15

    Aug 15, 2011 ... This study was conducted to investigate the effect of salt stress on germination of 28 red clover. (Trifolium pratense ... tolerance with the aim of improving crop plants (Zhu,. 2001) or soil .... The interaction of salinity and population in terms of PI ... in shoot growth is probably due to hormonal signals generated ...

  16. Genetic variations between two ecotypes of Egyptian clover by inter ...

    African Journals Online (AJOL)

    Aghomotsegin

    2015-06-10

    Jun 10, 2015 ... Four Egyptian clover (Trifolium alexandrinum L) cultivars representing two ecotypes were used in the present study. Fahl cultivar is prevalent in whole Egypt and is good for single cut as it has poor regeneration ability, whereas Serw1, Giza6 and Gemmiza1 give 5-6 cuts of good fodder. Techniques based ...

  17. Kinetics and optimization of red clover pulp drying

    Directory of Open Access Journals (Sweden)

    A. A. Shevtsov

    2013-01-01

    Full Text Available The kinetics of the drying process conducted with bagasse red clover on an experimental dry¬er superheated steam at atmospheric pressure in the active hydrodynamic regimes were study¬ing. The rational intervals of parameter changes were obtained. The problem of optimization, which allowed allocating the optimum range of variation of the input factors according to three criteria through compromise was solved.

  18. Sources of N2O in organic grass-clover pastures

    OpenAIRE

    Ambus, P.

    2002-01-01

    Organic farming practises, and in particular dairy production systems based on grass-clover pastures are becoming increasingly abundant within Danish agriculture. Grass-clover pastures may provide a mitigation option to reduce grassland nitrous oxide (N2O) emissions (Velthof et al. 1998). The objectives of this work was to examine the relationship between N2O emissions and transformations of inorganic N in organically managed grass-clover pastures of different ages. Results from the projec...

  19. Estimating grass-clover ratio variations caused by traffic intensities using image analysis

    DEFF Research Database (Denmark)

    Jørgensen, Rasmus Nyholm; Sørensen, Claus Grøn; Green, Ole

    Grass and especially clover have a negative yield response as a function of  traffic intensity.  Conventional grass-clover production for silage have high traffic intensity due to fertilizing with slurry, cutting the grass, rolling the grass into swaths, and collecting and chopping the grass...... to fulfill the aim [1]http://www.ruralni.gov.uk/index/publications/press_articles/dairy-2/role-of-clover.htm...

  20. Non-perturbative improvement of stout-smeared three flavour clover fermions

    Energy Technology Data Exchange (ETDEWEB)

    Cundy, N.; Goeckeler, M. [Regensburg Univ. (Germany). Inst. fuer Theoretische Physik; Horsley, R. [Edinburgh Univ. (GB). School of Physics and Astronomy] (and others)

    2009-01-15

    We discuss a 3-flavour lattice QCD action with clover improvement in which the fermion matrix has single level stout smearing for the hopping terms together with unsmeared links for the clover term. With the (tree-level) Symanzik improved gluon action this constitutes the Stout Link Non-perturbative Clover or SLiNC action. To cancel O(a) terms the clover term coefficient has to be tuned. We present here results of a non-perturbative determination of this coefficient using the Schroedinger functional and as a by-product a determination of the critical hopping parameter. Comparisons of the results are made with lowest order perturbation theory. (orig.)

  1. Fate in Soil of Flavonoids Released from White Clover (Trifolium repens L.)

    DEFF Research Database (Denmark)

    Carlsen, Sandra C. K.; Pedersen, Hans A.; Spliid, Niels H.

    2012-01-01

    the presence in soil of bioactive secondary metabolites from clover has received limited attention. In this paper we examine for the first time the release of flavonoids both from field-grown white clover and from soil-incorporated white clover plants of flavonoids, as analyzed by LC-MS/MS. The dominant...... flavonoid aglycones were formononetin, medicarpin, and kaempferol. Soil-incorporated white clover plants generated high concentrations of the glycosides kaempferol-Rha-Xyl-Gal and quercetin-Xyl-Gal. Substantial amounts of kaempferol persisted in the soil for days while the other compounds were degraded...

  2. Production of N2O in grass-clover pastures

    International Nuclear Information System (INIS)

    Carter, M.S.

    2005-09-01

    Agricultural soils are known to be a considerable source of the strong greenhouse gas nitrous oxide (N 2 O), and in soil N 2 O is mainly produced by nitrifying and denitrifying bacteria. In Denmark, grass-clover pastures are an important component of the cropping system in organic as well as conventional dairy farming, and on a European scale grass-clover mixtures represent a large part of the grazed grasslands. Biological dinitrogen (N 2 ) fixation in clover provides a major N input to these systems, but knowledge is sparse regarding the amount of fixed N 2 lost from the grasslands as N2O. Furthermore, urine patches deposited by grazing cattle are known to be hot-spots of N 2 O emission, but the mechanisms involved in the N 2 O production in urine-affected soil are very complex and not well understood. The aim of this Ph.D. project was to increase the knowledge of the biological and physical-chemical mechanisms, which control the production of N2O in grazed grass-clover pastures. Three experimental studies were conducted with the objectives of: 1: assessing the contribution of recently fixed N 2 as a source of N 2 O. 2: examining the link between N 2 O emission and carbon mineralization in urine patches. 3: investigating the effect of urine on the rates and N 2 O loss ratios of nitrification and denitrification, and evaluating the impact of the chemical conditions that arise in urine affected soil. The results revealed that only 3.2 ± 0.5 ppm of the recently fixed N 2 was emitted as N2O on a daily basis. Thus, recently fixed N released via easily degradable clover residues appears to be a minor source of N2O. Furthermore, increased N 2 O emission following urine application at rates up to 5.5 g N m -2 was not caused by enhanced denitrification stimulated by labile compounds released from scorched plant roots. Finally, the increase of soil pH and ammonium following urine application led to raised nitrification rate, which appeared to be the most important factor

  3. Cumulative effects of white clover residues on the changes in soil ...

    African Journals Online (AJOL)

    White clover grows naturally all over the Himalayan regions including the hilly areas of the state of Azad Jammu and Kashmir (AJK), Pakistan. This study was conducted to investigate the effects of white clover residues alone or in combination with phosphorus (P) fertilizer on maize (Zea mays L.) yield, nutrient uptake and ...

  4. Inclusion of caraway in the ryegrass-red clover mixture modifies soil microbial community composition

    DEFF Research Database (Denmark)

    Cong, Wenfeng; Jing, Jingying; Søegaard, Karen

    -containing grass-clover mixtures may potentially affect soil microbial community structure, biomass and associated ecosystem functions, but it is yet to be elucidated. We hypothesized that inclusion of plantain in the grass-clover mixture would enhance soil microbial biomas and functions through its high biomass...

  5. A New NPGS Special Collection: Norman L. Taylor University of Kentucky Clover Collection

    Science.gov (United States)

    Dr. Norman L. Taylor was a world renowned Professor and clover breeder in the Department of Plant and Soil Sciences at the University of Kentucky for 48 years. Following retirement in 2001, he continued working on clovers up until his death in 2010. Dr. Taylor’s entire career was devoted to enhancin...

  6. Linyphiid spider populations in sustainable wheat‐clover bi‐cropping compared to conventional wheat‐growing practice

    DEFF Research Database (Denmark)

    Gravesen, Eigil Vestergaard

    2008-01-01

    Linyphiid web densities in wheat-clover bi-crop systems where winter wheat was grown in an under-storey of white clover were compared with web densities estimated in conventional wheat-growing systems. The web densities in the wheat-clover bi-crop systems were on average between 200 and 250 webs ...

  7. Effects of clover density on N2O emissions and plant-soil N transfers in a fertilised upland pasture

    DEFF Research Database (Denmark)

    Klumpp, Katja; Bloor, Juliette M. G.; Ambus, Per

    2011-01-01

    regression analysis revealed that water-filled pore space (WFPS) and clover dry mass were the main factors driving cumulative N2O emissions in the high clover treatment, whereas variation in cumulated N2O emissions in the low clover treatment was best explained by WFPS and grass mass. We hypothesize...

  8. Specific activity isolation and determination of radioactive Estrogenic Substances in White Clover

    International Nuclear Information System (INIS)

    Pupiales T, G.; Mejia M, G.

    1986-01-01

    Due to high number of leguminous that exhibit estrogenic activity, subterranean clover between others, which causes infertility in sheep that eat it. It has been considered that white clover (Trifolium repens, variety Ladino, is an specie of low estrogenic activity, however at Bogota City (Colombia) it has high estrogenic activity and may cause reduction in the dairy cattle fertility. Research done in the IAN (today Ingeominas) over this clover variety, showed that the radioactivity substances presents in the white clover have high activity for stradiol, affecting organs from mouse females; Isoflavonoids from vegetables have an anabolism and utero tropic action; estrogenic activity of clover leaves, was exponentially proportional to the amount of ultraviolet radioactivity, falling upon plants during leaves development stage

  9. Honeybees tolerate cyanogenic glucosides from clover nectar and flowers

    DEFF Research Database (Denmark)

    Lecocq, Antoine; Green, Amelia Ann; Pinheiro de Castro, Érika Cristina

    2018-01-01

    Honeybees (Apis mellifera) pollinate flowers and collect nectar from many important crops. White clover (Trifolium repens) is widely grown as a temperate forage crop, and requires honeybee pollination for seed set. In this study, using a quantitative LC-MS (Liquid Chromatography-Mass Spectrometry...... indicates that plant secondary metabolites found in nectar may protect pollinators from disease or predators. In a laboratory survival study with chronic feeding of secondary metabolites, we show that honeybees can ingest the cyanogenic glucosides linamarin and amygdalin at naturally occurring...

  10. Towards integrated pest management in red clover seed production.

    Science.gov (United States)

    Lundin, Ola; Rundlöf, Maj; Smith, Henrik G; Bommarco, Riccardo

    2012-10-01

    The development of integrated pest management is hampered by lack of information on how insect pest abundances relate to yield losses, and how pests are affected by control measures. In this study, we develop integrated pest management tactics for Apion spp. weevils (Coleoptera: Brentidae) in seed production of red clover, Trifolium pratense L. We tested a method to forecast pest damage, quantified the relationship between pest abundance and yield, and evaluated chemical and biological pest control in 29 Swedish red clover fields in 2008 and 2011. Pest inflorescence abundance, which had a highly negative effect on yield, could be predicted with pan trap catches of adult pests. In 2008, chemical control with typically one application of pyrethroids was ineffective both in decreasing pest abundances and in increasing yields. In 2011, when chemical control included applications of the neonicotinoid thiacloprid, pest abundances decreased and yields increased considerably in treated field zones. A post hoc analysis indicated that using pyrethroids in addition to thiacloprid was largely redundant. Infestation rates by parasitoids was higher and reached average levels of around 40% in insecticide treated field zones in 2011, which is a level of interest for biological pest control. Based on the data presented, an economic threshold for chemical control is developed, and guidelines are provided on minimum effective chemical pest control.

  11. Identification of the S-transferase like superfamily bacillithiol transferases encoded by Bacillus subtilis

    Science.gov (United States)

    Perera, Varahenage R.; Lapek, John D.; Newton, Gerald L.; Gonzalez, David J.; Pogliano, Kit

    2018-01-01

    Bacillithiol is a low molecular weight thiol found in Firmicutes that is analogous to glutathione, which is absent in these bacteria. Bacillithiol transferases catalyze the transfer of bacillithiol to various substrates. The S-transferase-like (STL) superfamily contains over 30,000 putative members, including bacillithiol transferases. Proteins in this family are extremely divergent and are related by structural rather than sequence similarity, leaving it unclear if all share the same biochemical activity. Bacillus subtilis encodes eight predicted STL superfamily members, only one of which has been shown to be a bacillithiol transferase. Here we find that the seven remaining proteins show varying levels of metal dependent bacillithiol transferase activity. We have renamed the eight enzymes BstA-H. Mass spectrometry and gene expression studies revealed that all of the enzymes are produced to varying levels during growth and sporulation, with BstB and BstE being the most abundant and BstF and BstH being the least abundant. Interestingly, several bacillithiol transferases are induced in the mother cell during sporulation. A strain lacking all eight bacillithiol transferases showed normal growth in the presence of stressors that adversely affect growth of bacillithiol-deficient strains, such as paraquat and CdCl2. Thus, the STL bacillithiol transferases represent a new group of proteins that play currently unknown, but potentially significant roles in bacillithiol-dependent reactions. We conclude that these enzymes are highly divergent, perhaps to cope with an equally diverse array of endogenous or exogenous toxic metabolites and oxidants. PMID:29451913

  12. Does white clover (Trifolium repens abundance in temperate pastures determine Sitona obsoletus (Coleoptera: Curculionidae larval populations?

    Directory of Open Access Journals (Sweden)

    Mark Richard McNeill

    2016-09-01

    Full Text Available To determine if host plant abundance determined the size of clover root weevil (CRW Sitona obsoletus larval populations, a study was conducted over four years in plots sown in ryegrass (Lolium perenne (cv. Nui sown at either 6 or 30 kg/ha and white clover (Trifolium repens sown at a uniform rate of 8 kg/ha. This provided a range of % white clover content to investigate CRW population establishment and impacts on white clover survival. Larval sampling was carried out in spring (October when larval densities are near their spring peak at Lincoln (Canterbury, New Zealand with % clover measured in autumn (April and spring (September of each year. Overall, mean larval densities measured in spring 2012, 2013, 2014 and 2015 were 310, 38, 59 and 31 larvae m-2, respectively. There was a significant decline in larval populations between 2012 and 2013, but spring populations were relatively uniform thereafter. The mean % white clover measured in autumns of 2012 to 2015 was 17, 10, 3 and 11%, respectively. In comparison, mean spring % white clover from 2012 to 2015, averaged c. 5% each year. Analysis relating spring (October larval populations to % white clover measured in each plot in autumn (April found the 2012 larval population to be statistically significantly larger in the ryegrass 6 kg/ha plots than 30 kg/ha plots. Thereafter, sowing rate had no significant effect on larval populations. From 2013 to 2015, spring larval populations had a negative relationship with the previous autumn % white clover with the relationship highly significant for the 2014 data. When CRW larval populations in spring 2013 to 2015 were predicted from the 2013 to 2015 autumn % white clover, respectively, based on their positive relationship in 2012, the predicted densities were substantially larger than those observed. Conversely, when 2015 spring larval data and % clover was regressed against 2012-2014 larval populations, observed densities tended to be higher than predicted

  13. Does White Clover (Trifolium repens) Abundance in Temperate Pastures Determine Sitona obsoletus (Coleoptera: Curculionidae) Larval Populations?

    Science.gov (United States)

    McNeill, Mark R; van Koten, Chikako; Cave, Vanessa M; Chapman, David; Hodgson, Hamish

    2016-01-01

    To determine if host plant abundance determined the size of clover root weevil (CRW) Sitona obsoletus larval populations, a study was conducted over 4 years in plots sown in ryegrass ( Lolium perenne ) (cv. Nui) sown at either 6 or 30 kg/ha and white clover ( Trifolium repens ) sown at a uniform rate of 8 kg/ha. This provided a range of % white clover content to investigate CRW population establishment and impacts on white clover survival. Larval sampling was carried out in spring (October) when larval densities are near their spring peak at Lincoln (Canterbury, New Zealand) with % clover measured in autumn (April) and spring (September) of each year. Overall, mean larval densities measured in spring 2012-2015 were 310, 38, 59, and 31 larvae m -2 , respectively. There was a significant decline in larval populations between 2012 and 2013, but spring populations were relatively uniform thereafter. The mean % white clover measured in autumns of 2012 to 2015 was 17, 10, 3, and 11%, respectively. In comparison, mean spring % white clover from 2012 to 2015, averaged c. 5% each year. Analysis relating spring (October) larval populations to % white clover measured in each plot in autumn (April) found the 2012 larval population to be statistically significantly larger in the ryegrass 6 kg/ha plots than 30 kg/ha plots. Thereafter, sowing rate had no significant effect on larval populations. From 2013 to 2015, spring larval populations had a negative relationship with the previous autumn % white clover with the relationship highly significant for the 2014 data. When CRW larval populations in spring 2013 to 2015 were predicted from the 2013 to 2015 autumn % white clover, respectively, based on their positive relationship in 2012, the predicted densities were substantially larger than those observed. Conversely, when 2015 spring larval data and % clover was regressed against 2012-2014 larval populations, observed densities tended to be higher than predicted, but the numbers

  14. Leaching of cyanogenic glucosides and cyanide from white clover green manure

    DEFF Research Database (Denmark)

    Bjarnholt, Nanna; Lægdsmand, Mette; Hansen, Hans Chr. Bruun

    2008-01-01

    Use of crops for green manure as a substitute for chemical fertilizers and pesticides is an important approach towards more sustainable agricultural practices. Green manure from white clover is rich in nitrogen but white clover also produces the cyanogenic glucosides (CGs) linamarin...... and lotaustralin; CGs release toxic hydrogen cyanide (HCN) upon hydrolysis which may be utilized for pest control. We demonstrate that applying CGs in the form of a liquid extract of white clover to large columns of intact agricultural soils can result in leaching of toxic cyanide species to a depth of at least 1...

  15. Report on the intercomparison run IAEA-156 radionuclides in clover

    International Nuclear Information System (INIS)

    Strachnov, V.; Valkovic, V.; Dekner, R.

    1991-01-01

    This report contains the results of the intercomparison IAEA-156 on the determination of radionuclides in clover. Initially participants were requested to determine the levels of 134 Cs, 137 Cs, 40 K, 90 Sr and invited to provide data for other radionuclides. The participants included 46 laboratories located in 25 countries, and statistical evaluation of their data yield recommended values for these four radionuclides. Additional radionuclides reported were 210 Pb, 239 Pu and 125 Sb; however, insufficient data exists to statistically determine recommended values for these radionuclides. The following are the recommended values, with confidence intervals, for the most frequently measured radionuclides (reference date: 1 August 1986): 134 Cs 132.1 Bq/kg (126.4-137.7); 137 Cs 264 Bq/kg (254-274); 40 K 657 Bq/kg (637-676); 90 Sr 14.8 Bq/kg (13.4-16.3). Figs and tabs

  16. Relationship between nitrogen cycling and nitrous oxide emission in grass-clover pasture

    OpenAIRE

    Ambus, P.

    2005-01-01

    The paper reports on a work assessing the relationship between gross N transformations in grass-clover soils and emissions of nitrous oxide. By this manner, the source strength of the biogenic processes responsible for nitrous oxide production is evaluated.

  17. N transfer in three species grass-clover mixtures with chicory, ribwort plantain or caraway

    DEFF Research Database (Denmark)

    Dhamala, Nawa Raj; Rasmussen, Jim; Carlsson, Georg

    2017-01-01

    Background and aimsThere is substantial evidence that legume-derived Nitrogen (N) is transferred to neighboring non-legumes in grassland mixtures. However, there is sparse information about how deep rooted non-legume forage herbs (forbs) influence N transfer in multi-species grasslands. Methodology......Red clover (Trifolium pretense L.) was grown together with perennial ryegrass (Lolium perenne L.) and one of three forb species: chicory (Cichorium intybus L.), ribwort plantain (Plantago lanceolata L.) or caraway (Carum carvi L.) in a field experiment. During the first year after the establishment, red...... clover leaves were labeled with 15N-urea to determine the N transfer from red clover to companion ryegrass and forbs. ResultsOn an annual basis, up to 15 % of red clover N was transferred to the companion ryegrass and forbs, but predominantly to the grass. The forb species did not differ in their ability...

  18. Reduction in clover-grass yield caused by different traffic intensities

    DEFF Research Database (Denmark)

    Green, Ole; Jørgensen, Rasmus Nyholm; Kristensen, Kristian

    Different traffic intensities have been shown to have a negative influence on the yield of grass and clover. A full scale grass-clover field trial was established to estimate the effect on clover-grass yields as a function of different wheel loads and tire pressures. The trial comprised 16...... close to the north, south and east border of the field. No significant interactions were found between the timing of crop and soil damage as affected by wheel load and tire pressure. However, at specific times, there was a significant effect of wheel load and secondary by the tire pressure. At all...... measurement times, the yield was lower using a wheel load of 4745 kg than for a wheel load of 2865 kg.     Key words (for Electronic Reference Library) Traffic intensities, tire load/pressure, clover/grass, yield loss, ...

  19. SIKLODEKSTRIN GLIKOSIL TRANSFERASE DAN PEMANFAATANNYA DALAM INDUSTRI [Cyclodextrin Glycosyl Transferase and its application in industries

    Directory of Open Access Journals (Sweden)

    Budiasih Wahyuntari

    2005-12-01

    Full Text Available Cyclodextrin glycosyl transferase (CGT-ase is mainly produced by Bacilli. Systematical name of the enzyme is E.C. 2.4.1.19 a-1,4 glucan-4-glycosyl transferase. The enzyme catalyzes hydrolysis of starch intramolecular, and intermolecular transglycosylation of a-1,4, glucan chains. Cyclodextrins are a-1,4 linked cyclic oligosaccharides resulting from enzymatic degradation of starch by cyclodextrin glycosyl transferase through untramolecular transglycosylation. The major cyclodextrins are made up of 6, 7 and 8 glucopyranose units which are known as a-, b-, and y-cyclodextrin. All CGT-ase catalyze three kinds of cyclodextrins, the proportion of the cyclodextrins depends on the enzyme source and reaction conditions. The intermolecular transglycosylation ability of the enzyme has been applied in transfering glycosyl residues into suitable acceptor. Transglycosylation by the enzymes have been tested to improve solubility of some flavonoids and to favor precipitation ci some glycosides.

  20. Exogenous Application of GABA Improves PEG-Induced Drought Tolerance Positively Associated with GABA-Shunt, Polyamines, and Proline Metabolism in White Clover.

    Science.gov (United States)

    Yong, Bin; Xie, Huan; Li, Zhou; Li, Ya-Ping; Zhang, Yan; Nie, Gang; Zhang, Xin-Quan; Ma, Xiao; Huang, Lin-Kai; Yan, Yan-Hong; Peng, Yan

    2017-01-01

    In order to investigate the physiological effects of exogenous γ-aminobutyric acid (GABA) on drought tolerance in white clover (Trifolium repens), GABA shunt, polyamines (PAs), and proline (Pro) metabolism were examined after plants pretreated with or without GABA (8 mM) and then exposed to water or 15% PEG-induced drought stress in growth chamber. In this study, exogenous application of GABA effectively alleviated drought-induced damage in leaves, as reflected by significantly higher relative water content, lower electrolyte leakage, lipid peroxidation, and leaf wilt. Exogenous GABA further promoted drought-induced increases in GABA transaminase and alpha ketone glutarate dehydrogenase activities, but inhibited glutamate decarboxylase activity under both control and drought conditions, resulting in an increase in endogenous glutamate (Glu) and GABA content. Besides, exogenous GABA could well accelerated PAs synthesis and suppressed PAs catabolism, which lead to the extremely enhanced different types of PAs content (free Put and Spd, insoluble bound Spd and Spm, soluble conjugated Spd and Spm, and total Put, Spd and Spm) under drought stress. In addition, exogenous GABA application further activated drought-induced Δ 1 -pyrroline-5-carboxylate synthetase and proline dehydrogenase activities, but suppressed drought-facilitated ornithine -δ-amino transferase activities, leading to a higher Pro accumulation and metabolism in GABA-pretreated plants in the middle and last period of drought. The results suggested that increased endogenous GABA by exogenous GABA treatment could improve drought tolerance of white clover associated with a positive regulation in the GABA-shunt, PAs and Pro metabolism.

  1. Effect of toasting field beans and of grass-clover

    DEFF Research Database (Denmark)

    Mogensen, Lisbeth; Vestergaard, Jannie Steensig; Fretté, Xavier

    2010-01-01

    The effect of toasting field beans and of grass-clover: maize silage ratio on milk production, milk composition and the sensory quality of the milk was investigated in a 2   2 factorial experiment. Toasting of field beans resulted in lower milk contents of both fat (44.2 versus 46.1 g/kg, P = 0.......02) and protein (33.5 versus 34.2 g/kg, P = 0.008), whereas milk production, urea and somatic cell contents were unaffected compared with the untreated field beans. Increasing the proportion of maize silage (from 9 to 21% of DM) in the ration decreased the content of urea in milk (P = 0.002), whereas milk......-β-carotene (P = 0.04) and β-carotene (P = 0.05). Toasting of field beans compared with untreated field beans did not affect the milk content of carotenoids and had only small effects on fatty acid composition. Regarding the sensory quality, the four treatments resulted in milk being characterized...

  2. Nitrous oxide emissions from clover in the Mediterranean environment

    Directory of Open Access Journals (Sweden)

    Iride Volpi

    2016-06-01

    Full Text Available Introducing nitrogen N2-fixing crops into cereal-based crop rotations reduces N-fertiliser use and may mitigate soil emissions of nitrous oxide (N2O. However, the effect of the cultivation of N2-fixing crops on N2O emissions is still not well understood. N2O from N2-fixing crops can be emitted in two ways: during biological N2 fixation itself and when legume residues are returned to the soil. A field trial was carried out on clover (Trifolium squarrosum Savi to test the role of leguminous crops on N2O emissions in the Mediterranean environment. Monitoring was performed from December 2013 to September 2014. Cumulated N-N2O fluxes were calculated for the growing season (Phase 1 and the post-harvest period (Phase 2 in order to assess the importance of each phase. Our results did not show statistically significant differences between the two phases in term of contribution to the total cumulative N-N2O emissions, in fact Phase 1 and Phase 2 accounted respectively for 43 and 57% of the total.

  3. Elemental analysis and nutritional value of edible Trifolium (clover) species.

    Science.gov (United States)

    Gounden, Thaveshan; Moodley, Roshila; Jonnalagadda, Sreekantha B

    2018-04-30

    Trifolium species, commonly known as clover species, have a cosmopolitan distribution and, as such, are used in many different traditional systems of medicine and consumed by many communities all over the world. In this study, the elemental distribution and nutritional value of five edible Trifolium species, namely, Trifolium africanum, Trifolium burchellianum, Trifolium repens, Trifolium dubium and Trifolium pratense were investigated to evaluate the potential of these plant species to alleviate malnutrition, thereby contributing toward the fight against food insecurity. Trifolium species were found to be a rich alternate source of essential nutrients with concentrations of elements being in decreasing order of Ca > Mg > Fe > Mn > Zn > Se > Cu > Cr > Pb > Ni > Co > Cd > As and with adequate levels of lipids (4.2 to 8.6%), proteins (35.1 to 45.4%) and carbohydrates (26.7 to 47.0%). Trifolium species were found to be rich in Se (contributing greater than 516% toward its RDA) with T. dubium having a concentration of 0.53 mg 10 g -1 , dry mass, which is higher than Brazil nuts. T. pratense was found to be the most suitable species for human consumption due to it having low levels of toxic metals (As, Cd and Pb) while being rich in macro- and micro-elements, especially Fe (7.84 mg 10 g -1 , dry mass) and Se (0.36 mg 10 g -1 , dry mass).

  4. Comparison of breeding methods for forage yield in red clover

    Directory of Open Access Journals (Sweden)

    Libor Jalůvka

    2009-01-01

    Full Text Available Three methods of red clover (Trifolium pratense L. breeding for forage yield in two harvest years on locations in Bredelokke (Denmark, Hladké Životice (Czech Republic and Les Alleuds (France were compared. Three types of 46 candivars1, developed by A recurrent selection in subsequent generations (37 candivars, divided into early and late group, B polycross progenies (4 candivars and C ge­no-phe­no­ty­pic selection (5 candivars were compared. The trials were sown in 2005 and cut three times in 2006 and 2007; their evaluation is based primarily on total yield of dry matter. The candivars developed by polycross and geno-phenotypic selections gave significantly higher yields than candivars from the recurrent selection. However, the candivars developed by the methods B and C did not differ significantly. The candivars developed by these progressive methods were suitable for higher yielding and drier environment in Hladké Životice (where was the highest yield level even if averaged annual precipitation were lower by 73 and 113 mm in comparison to other locations, respectively; here was ave­ra­ge yield higher by 19 and 13% for B and C in comparison to A method. Highly significant interaction of the candivars with locations was found. It can be concluded that varieties specifically aimed to different locations by the methods B and C should be bred; also the parental entries should be selected there.

  5. Assessment of nickel bioavailability through chemical extractants and red clover (Trifolium pratense L.) in an amended soil: Related changes in various parameters of red clover.

    Science.gov (United States)

    Shahbaz, Ali Khan; Iqbal, Muhammad; Jabbar, Abdul; Hussain, Sabir; Ibrahim, Muhammad

    2018-03-01

    Application of immobilizing agents may efficiently reduce the bioavailability of nickel (Ni) in the soil. Here we report the effect of biochar (BC), gravel sludge (GS) and zeolite (ZE) as a sole treatment and their combinations on the bioavailability of Ni after their application into a Ni-polluted soil. The bioavailability of Ni after the application of immobilizing agents was assessed through an indicator plant (red clover) and chemical indicators of bioavailability like soil water extract (SWE), DTPA and Ca(NO 3 ) 2 extracts. Additionally, the effects of Ni bioavailability and immobilizing agents on the growth, physiological and biochemical attributes of red clover were also observed. Application of ZE significantly reduced Ni concentrations in all chemical extracts compared to rest of the treatments. Similarly, the combined application of BC and ZE (BC+ ZE) significantly reduced Ni concentrations, reactive oxygen species (ROS) whereas, significant enhancement in the growth, physiological and biochemical attributes along with an improvement in antioxidant defence machinery of red clover plant, compared to rest of the treatments, were observed. Furthermore, BC+ ZE treatment significantly reduced bioconcentration factor (BCF) and bioaccumulation factor (BAF) of Ni in red clover, compared to rest of the treatments. The Ni concentrations in red clover leaves individually reflected a good correlation with Ni concentrations in the extracts (SWE at R 2 =0.79, DTPA extract at R 2 =0.84 and Ca(NO 3 ) 2 extracts at R 2 =0.86). Our results indicate that combined application of ZE and BC can significantly reduce the Ni bioavailability in the soil while in parallel improve the antioxidant defence mechanism in plants. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Transformation kinetics of corn and clover residues in mineral substrates of different composition

    Science.gov (United States)

    Pinskii, D. L.; Maltseva, A. N.; Zolotareva, B. N.; Dmitrieva, E. D.

    2017-06-01

    Mineralization kinetics of corn and clover residues in quartz sand, loam, sand + 15% bentonite, and sand + 30% kaolinite have been studied. A scheme has been proposed for the transformation of plant residues in mineral substrates. Kinetic parameters of mineralization have been calculated with the use of a first-order two-term exponential polynomial. It has been shown that the share of labile organic carbon pool in the clover biomass is higher (57-63%) than in the corn biomass (47-49%), which is related to the biochemical composition of plant residues. The mineralization constants of clover residues generally significantly exceed those of corn because of the stronger stabilization of the decomposition products of corn residues. The turnover time of the labile clover pool (4-9 days) in all substrates and that of the labile corn pool (8-10 days) in sands and substrates containing kaolinites and bentonite are typical for organic acids, amino acids, and simple sugars. In the loamy substrate, the turnover time of labile corn pool is about 46 days due to the stronger stabilization of components of the labile pool containing large amounts of organic acids. The turnover time of the stable clover pool (0.95 years) is significantly lower than that of the stable corn pool (1.60 years) and largely corresponds to the turnover time of plant biomass.

  7. Phenotypic and genetic characterization of wildland collections of western and Searls prairie clovers for rangeland revegetation in the western USA

    Science.gov (United States)

    Kishor Bhattarai

    2010-01-01

    Western prairie clover [Dalea ornata (Douglas ex Hook.) Eaton & J. Wright] is a perennial legume that occurs in the northern Great Basin, Snake River Basin, and southern Columbia Plateau, whereas Searls prairie clover [Dalea searlsiae (A. Gray) Barneby], also a perennial legume, occurs in the southern Great Basin and surrounding areas. Understanding the genetic and...

  8. Phyto-oestrogens in herbage and milk from cows grazing whiteclover, red clover, lucerne or chicory-rich pastures

    DEFF Research Database (Denmark)

    Andersen, C; Nielsen, T S; Purup, S

    2009-01-01

    A grazing experiment was carried out to study the concentration of phyto-oestrogens in herbage for cattle and in milk during two periods (May and June). Forty-eight Danish Holstein cows were divided into four groups with four treatment diets; white clover, red clover, lucerne and chicory-rich pas...

  9. Effects of field characteristics on abundance of bumblebees (Bombus spp.) and seed yield in red clover fields

    DEFF Research Database (Denmark)

    Wermuth, Kirsten Haugaard; Dupont, Yoko L.

    2010-01-01

    Red clover is a key floral ressource for bumblebees (Bombus spp.).We here investigate variation within and among red clover fields in species richness and abundance of Bombus spp. in addition to Apis mellifera. Bumblebee individuals were grouped into the following functional groups, based on castes...

  10. A survey of the invertebrates feeding on living clover roots (Trifolium repens L.) using 32P as a radiotracer

    International Nuclear Information System (INIS)

    Baylis, J.P.; Cherrett, J.M.; Ford, J.B.

    1986-01-01

    Clover roots were labelled in the field with 32 P and the radioactive soil fauna were detected by autoradiography. The animals which consumed labelled clover roots were, in order of importance, earthworms (Aporrectodea longa, A. caliginosa and Lumbricus rubellus), weevil larvae, dipteran larvae (Bibio marci) and a few Collembola (family: Entomobryidae). (author)

  11. 7 CFR 202.44 - Proceedings under section 305(b) to determine whether foreign alfalfa or red clover seed is not...

    Science.gov (United States)

    2010-01-01

    ... foreign alfalfa or red clover seed is not adapted for general agricultural use in the United States. 202... Proceedings under section 305(b) to determine whether foreign alfalfa or red clover seed is not adapted for... for the purpose of determining whether seed of alfalfa or red clover from any foreign country or...

  12. Nitrogen fertilization of grass/clover swards under cutting or grazing by dairy cows

    DEFF Research Database (Denmark)

    Søegaard, Karen

    2009-01-01

    farms over a period of three years. Nitrogen was applied at four rates (0, 75, 150, and 225 kg N year-1) with cutting or grazing regime in Year 1 and Year 2, after establishment. A spring-only application of 150 kg N was compared with four applications during the season, which was the fertilization...... affected. The results indicate different possibilities for strategic fertilization both at farm and field level, and in swards with a high clover content it demonstrates how the clover content can be used as a buffer both for maximizing the N-response and for manipulating the production profile.......Intensively managed perennial ryegrass/white clover (Lolium perenne L. and Trifolium repens L.) swards receive relatively high levels of fertilizer N, and high N surpluses can subsequently be found. The N-fertilization effects on growth, yield, and herbage quality were therefore examined on three...

  13. Analysis of the effects of pair production for the suppressed clover detector

    International Nuclear Information System (INIS)

    Kshetri, R.

    2014-01-01

    Full energy peak, single escape peak and double escape peak areas have been extracted for an escape suppressed clover detector. Results have been obtained for the single crystal and addback modes of operation as well as the active and passive suppression cases at several gamma energies. We have compared the ratio of single escape peak areas in addback mode with that of single crystal mode to study if the single escape peak gains or loses counts due to addback mode. Detailed analysis has been performed for quantifying the advantages of using addback mode and active suppression. Comparison is made for different types of clover detectors with different volumes

  14. Effect of red clover on the microbial transformation of phenanthrene and octadecane in the soil

    Science.gov (United States)

    Nazarov, A. V.; Shestakova, E. A.; Anan'yina, L. N.

    2017-08-01

    The influence of red clover ( Trifolium pratense L.) plants on the decomposition of phenanthrene and octadecane in the soil has been studied. Effect of plant root mass on the abundance of hydrocarbondegrading bacteria, the composition of their communities, and the degradation of hydrocarbons in the soil has been revealed. Changes in the taxonomic composition of hydrocarbon-degrading bacteria under the effect of red clover are characterized by an increase in the abundance of species from the genera Acinetobacter, Kaistia, Novosphingobium, Pseudomonas, and Xanthomonas. A positive effect of the studied microbial-plant association on the degradation of octadecane and especially phenanthrene in the soil has been revealed.

  15. Enchytraeids as indicator of soil quality in temporary organic grass-clover leys under contrasting management

    DEFF Research Database (Denmark)

    Maraldo, Kristine; Schmelz, Rüdiger; Larsen, Thomas

    2015-01-01

    One objective in organic farming is to sustain the quality of the soil resource. Because enchytraeids are an important soil faunal component, they stand as bioindicators of soil quality. We tested this candidature in a field experiment on loamy sand soil with 1- and 4-year old grass-clover leys...... interactions among soil physical, chemical and biological properties suggest that enchytraeid abundance is not a feasible stand-alone indicator of management impacts on soil quality in temporary grass-clover leys but may candidate as one of several biological key parameters in more comprehensive soil quality...

  16. Preliminary research on amino acid composition and nutritional value of clover proteins

    Directory of Open Access Journals (Sweden)

    L. Kłyszejko-Stefanowicz

    2015-06-01

    Full Text Available The amino acid composition and nutritional value of 5 clover varieties including 3 Polish ones ('Gloria', 'Hruszowska', 'Skrzeszowicka' and 2 of foreign origin ('Rotra' and 'Violetta' were investigated. No significant differences in the total protein content (19.2–20.0% of dry matter as well as in qualitative amino acid composition were found among the clover varieties under examination. EAA index (Essential amino acid index calculated according to Oser for 'Gloria' and 'Hruszowska' showed the highest nutritional value was – 40. The lowest value of EAA index was found for 'Violetta' cvar. – 32, intermediate values however for Rotra and Skrzeszowicka was 37 and 36.

  17. Estimation of the Botanical Composition of Clover-Grass Leys from RGB Images Using Data Simulation and Fully Convolutional Neural Networks

    Science.gov (United States)

    Steen, Kim Arild; Green, Ole; Karstoft, Henrik

    2017-01-01

    Optimal fertilization of clover-grass fields relies on knowledge of the clover and grass fractions. This study shows how knowledge can be obtained by analyzing images collected in fields automatically. A fully convolutional neural network was trained to create a pixel-wise classification of clover, grass, and weeds in red, green, and blue (RGB) images of clover-grass mixtures. The estimated clover fractions of the dry matter from the images were found to be highly correlated with the real clover fractions of the dry matter, making this a cheap and non-destructive way of monitoring clover-grass fields. The network was trained solely on simulated top-down images of clover-grass fields. This enables the network to distinguish clover, grass, and weed pixels in real images. The use of simulated images for training reduces the manual labor to a few hours, as compared to more than 3000 h when all the real images are annotated for training. The network was tested on images with varied clover/grass ratios and achieved an overall pixel classification accuracy of 83.4%, while estimating the dry matter clover fraction with a standard deviation of 7.8%. PMID:29258215

  18. Genomic organization of plant aminopropyl transferases.

    Science.gov (United States)

    Rodríguez-Kessler, Margarita; Delgado-Sánchez, Pablo; Rodríguez-Kessler, Gabriela Theresia; Moriguchi, Takaya; Jiménez-Bremont, Juan Francisco

    2010-07-01

    Aminopropyl transferases like spermidine synthase (SPDS; EC 2.5.1.16), spermine synthase and thermospermine synthase (SPMS, tSPMS; EC 2.5.1.22) belong to a class of widely distributed enzymes that use decarboxylated S-adenosylmethionine as an aminopropyl donor and putrescine or spermidine as an amino acceptor to form in that order spermidine, spermine or thermospermine. We describe the analysis of plant genomic sequences encoding SPDS, SPMS, tSPMS and PMT (putrescine N-methyltransferase; EC 2.1.1.53). Genome organization (including exon size, gain and loss, as well as intron number, size, loss, retention, placement and phase, and the presence of transposons) of plant aminopropyl transferase genes were compared between the genomic sequences of SPDS, SPMS and tSPMS from Zea mays, Oryza sativa, Malus x domestica, Populus trichocarpa, Arabidopsis thaliana and Physcomitrella patens. In addition, the genomic organization of plant PMT genes, proposed to be derived from SPDS during the evolution of alkaloid metabolism, is illustrated. Herein, a particular conservation and arrangement of exon and intron sequences between plant SPDS, SPMS and PMT genes that clearly differs with that of ACL5 genes, is shown. The possible acquisition of the plant SPMS exon II and, in particular exon XI in the monocot SPMS genes, is a remarkable feature that allows their differentiation from SPDS genes. In accordance with our in silico analysis, functional complementation experiments of the maize ZmSPMS1 enzyme (previously considered to be SPDS) in yeast demonstrated its spermine synthase activity. Another significant aspect is the conservation of intron sequences among SPDS and PMT paralogs. In addition the existence of microsynteny among some SPDS paralogs, especially in P. trichocarpa and A. thaliana, supports duplication events of plant SPDS genes. Based in our analysis, we hypothesize that SPMS genes appeared with the divergence of vascular plants by a processes of gene duplication and the

  19. In situ carbon and nitrogen dynamics in ryegrass-clover mixtures

    DEFF Research Database (Denmark)

    Rasmussen, J.; Eriksen, J.; Jensen, Erik Steen

    2007-01-01

    =9). 15N-enriched compounds were not detected in percolating pore water, which may be caused by either dilution from irrigation or low availability of leachable N compounds. 14C was found solely as 14CO2 in the pore water indicating that dissolved organic carbon (DOC) did not originate from fresh......Carbon (C) and nitrogen (N) dynamics in a third production year ryegrass–clover mixture were investigated in the field. Cylinders (diameter 29.7 cm) were installed to depths of 20, 40 and 60 cm and equipped with suction cups to collect percolating pore water. Ryegrass and clover leaves were cross......-labelled with 14C- and 15N-enriched urea and the fate of the two tracers was studied for 3 months during summer. Transfer of 14C occurred mainly from ryegrass to clover, whereas the largest transfer of 15N was in the opposite direction. The average transfer of N from clover was 40% (SE±3.1, n=9) of N in ryegrass...

  20. Analysis of the soil food web structure under grass and grass clover

    NARCIS (Netherlands)

    Eekeren, van N.J.M.; Smeding, F.W.; Vries, de F.T.; Bloem, J.

    2006-01-01

    The below ground biodiversity of soil organisms plays an important role in the functioning of the the soil ecosystem, and consequently the above ground plant production. The objective of this study is to investigate the effect of grass or grass-clover in combination with fertilisation on the soil

  1. Cumulative effects of white clover residues on the changes in soil ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-18

    May 18, 2009 ... The residual effects of white clover on soil properties indicated a significant increase in saturation percentage (17–23%) and .... determined by leaching the soil with KCl followed by extraction of ...... accumulation, and oil quality of French basil. ... of the Final Workshop of the Farming System Integrated Pest.

  2. Improving Resilience of Northern Field Crop Systems Using Inter-Seeded Red Clover: A Review

    Directory of Open Access Journals (Sweden)

    William Deen

    2013-02-01

    Full Text Available In light of the environmental challenges ahead, resilience of the most abundant field crop production systems must be improved to guarantee yield stability with more efficient use of nitrogen inputs, soil and water resources. Along with genetic and agronomic innovations, diversification of northern agro-ecosystems using inter-seeded legumes provides further opportunities to improve land management practices that sustain crop yields and their resilience to biotic and abiotic stresses. Benefits of legume cover crops have been known for decades and red clover (Trifolium pratense is one of the most common and beneficial when frost-seeded under winter wheat in advance of maize in a rotation. However, its use has been declining mostly due to the use of synthetic fertilizers and herbicides, concerns over competition with the main crop and the inability to fully capture red clover benefits due to difficulties in the persistence of uniform stands. In this manuscript, we first review the environmental, agronomic, rotational and economical benefits associated with inter-seeded red clover. Red clover adaptation to a wide array of common wheat-based rotations, its potential to mitigate the effects of land degradation in a changing climate and its integration into sustainable food production systems are discussed. We then identify areas of research with significant potential to impact cropping system profitability and sustainability.

  3. Effects of titanium dioxide nanoparticles on red clover and its rhizobial symbiont

    NARCIS (Netherlands)

    Moll, Janine; Okupnik, Annette; Gogos, Alexander; Knauer, Katja; Bucheli, Thomas D.; Van Der Heijden, Marcel G A; Widmer, Franco

    2016-01-01

    Titanium dioxide nanoparticles (TiO2 NPs) are in consideration to be used in plant protection products. Before these products can be placed on the market, ecotoxicological tests have to be performed. In this study, the nitrogen fixing bacterium Rhizobium trifolii and red clover were exposed to two

  4. Biochar application rate affects biological nitrogen fixation in red clover conditional on potassium availability

    NARCIS (Netherlands)

    Mia, S.; van Groeningen, J.W.; Van de Voorde, T.F.J.; Oram, N.J.; Bezemer, T.M.; Mommer, Liesje; Jeffery, S.

    2014-01-01

    Increased biological nitrogen fixation (BNF) by legumes has been reported following biochar application to soils, but the mechanisms behind this phenomenon remain poorly elucidated. We investigated the effects of different biochar application rates on BNF in red clover (Trifolium pratense L.). Red

  5. Phenotypic and genetic characterization of western prairie clover collections from the western USA

    Science.gov (United States)

    Kishor Bhattarai; B. Shaun Bushman; Douglas A. Johnson; John G. Carman

    2010-01-01

    Few North American legumes are available for rangeland revegetation in the semiarid western United States. Western prairie clover (Dalea ornata [Douglas ex Hook.] Eaton & J. Wright) is a perennial legume with desirable forage characteristics and is distributed in the northern Great Basin, Snake River Basin, and southern Columbia Plateau. Understanding the...

  6. Searls prairie clover (Dalea searlsiae) for rangeland revegetation: Phenotypic and genetic evaluations

    Science.gov (United States)

    Kishor Bhattarai; Shaun Bushman; Douglas A. Johnson; John G. Carman

    2011-01-01

    Few North American legumes are available for use in rangeland revegetation in the western USA, but Searls prairie clover [Dalea searlsiae (A. Gray) Barneby] is one that holds promise. Commercial-scale seed production of this species could address the issues of unreliable seed availability and high seed costs associated with its wildland seed collection. To evaluate its...

  7. Molecular evolution of the Li/li chemical defence polymorphism in white clover (Trifolium repens L.).

    Science.gov (United States)

    Olsen, K M; Sutherland, B L; Small, L L

    2007-10-01

    White clover (Trifolium repens) is naturally polymorphic for cyanogenesis (hydrogen cyanide release following tissue damage). The ecological factors favouring cyanogenic and acyanogenic plants have been examined in numerous studies over the last half century, making this one of the best-documented examples of an adaptive polymorphism in plants. White clover cyanogenesis is controlled by two, independently segregating Mendelian genes: Ac/ac controls the presence/absence of cyanogenic glucosides; and Li/li controls the presence/absence of their hydrolysing enzyme, linamarase. In this study, we examine the molecular evolution and population genetics of Li as it relates to the cyanogenesis polymorphism. We report here that Li exists as a single-copy gene in plants possessing linamarase activity, and that the absence of enzyme activity in li/li plants is correlated with the absence of much or all of the gene from the white clover genome. Consistent with this finding, we confirm by reverse transcription-polymerase chain reaction that Li gene expression is absent in plants lacking enzyme activity. In a molecular population genetic analysis of Li and three unlinked genes using a worldwide sample of clover plants, we find an absence of nucleotide variation and statistically significant deviations from neutrality at Li; these findings are consistent with recent positive directional selection at this cyanogenesis locus.

  8. Forbs enhance productivity of unfertilised grass-clover leys and support low-carbon bioenergy

    DEFF Research Database (Denmark)

    Cong, Wen-Feng; Jing, Jingying; Rasmussen, Jim

    2017-01-01

    Intensively managed grasslands are dominated by highly productive grass-clover mixtures. Increasing crop diversity by inclusion of competitive forbs may enhance biomass production and sustainable biofuel production. Here we examined if one or all of three forbs (chicory, Cichorium intybus L.; car...

  9. Relationships between Nitrate and Oxygen Supply in Symbiotic Nitrogen Fixation by White Clover

    DEFF Research Database (Denmark)

    Minchin, F. R.; Ines Minguez, M.; Sheedy, J. E.

    1986-01-01

    Exposure of mature, nodulated plants of white clover (Trifolium repens) cv. Blanca to 330 mg dm−3 NO3-N for 8 d caused nitrogenase activity per plant to decrease by 80%. Total nodulated root respiration was not significantly affected but analysis of its components showed an 81% decrease in nitrog...

  10. Meat goat kids finished on alfalfa, red clover, or orchardgrass pastures: Carcass merit and meat quality

    Science.gov (United States)

    This experiment was conducted in 2005-2007 to evaluate carcass and meat quality parameters when meat goat kids were finished on alfalfa (Medicago sativa L; ALF); red clover (Trifolium pretense L.; RCG); or orchardgrass (Dactylis glomerata L; OGR) pastures. Final shrunk body weights were similar whe...

  11. The Genetic Architecture of Murine Glutathione Transferases.

    Directory of Open Access Journals (Sweden)

    Lu Lu

    Full Text Available Glutathione S-transferase (GST genes play a protective role against oxidative stress and may influence disease risk and drug pharmacokinetics. In this study, massive multiscalar trait profiling across a large population of mice derived from a cross between C57BL/6J (B6 and DBA2/J (D2--the BXD family--was combined with linkage and bioinformatic analyses to characterize mechanisms controlling GST expression and to identify downstream consequences of this variation. Similar to humans, mice show a wide range in expression of GST family members. Variation in the expression of Gsta4, Gstt2, Gstz1, Gsto1, and Mgst3 is modulated by local expression QTLs (eQTLs in several tissues. Higher expression of Gsto1 in brain and liver of BXD strains is strongly associated (P < 0.01 with inheritance of the B6 parental allele whereas higher expression of Gsta4 and Mgst3 in brain and liver, and Gstt2 and Gstz1 in brain is strongly associated with inheritance of the D2 parental allele. Allele-specific assays confirmed that expression of Gsto1, Gsta4, and Mgst3 are modulated by sequence variants within or near each gene locus. We exploited this endogenous variation to identify coexpression networks and downstream targets in mouse and human. Through a combined systems genetics approach, we provide new insight into the biological role of naturally occurring variants in GST genes.

  12. White clover fractions as protein source for monogastrics - Dry matter digestibility and Protein Digestibility-Corrected Amino Acid Scores

    DEFF Research Database (Denmark)

    Stødkilde, Lene; Damborg, Vinni K; Jørgensen, Henry

    2018-01-01

    BACKGROUND: The aim was to evaluate white clover as an alternative protein source for monogastrics. White clover plant and leaves were processed using a screw-press resulting in a solid pulp and a juice from which protein was acid-precipitated. The chemical composition of all fractions...... was determined and digestibility of dry matter (DM) and protein was assessed in an experiment with growing rats. RESULTS: Protein concentrates were produced with crude protein (CP) content of 451 g/kg DM and 530 g/kg DM for white clover plant and leaves, respectively and a pulp with CP content of 313 and 374 g...

  13. Glutathione transferase mimics : Micellar catalysis of an enzymic reaction

    NARCIS (Netherlands)

    Lindkvist, Björn; Weinander, Rolf; Engman, Lars; Koetse, Marc; Engberts, Jan B.F.N.; Morgenstern, Ralf

    1997-01-01

    Substances that mimic the enzyme action of glutathione transferases (which serve in detoxification) are described. These micellar catalysts enhance the reaction rate between thiols and activated halogenated nitroarenes as well as alpha,beta-unsaturated carbonyls. The nucleophilic aromatic

  14. The association between glutathione S-transferase P1 ...

    African Journals Online (AJOL)

    Mahmoud I. Mahmoud

    2011-08-10

    Aug 10, 2011 ... B-adrenergic receptor polymorphisms and response to salmeterol. Am J Respir Crit Care ... transferase Pi locus and association with susceptibility to bladder, testicular and prostate cancer. Carcinogenesis 1997;18(4):641–4.

  15. Effect of fertilizer type on cadmium and fluorine concentrations in clover herbage

    International Nuclear Information System (INIS)

    McLaughlin, M.J.

    2002-01-01

    This study investigated whether changing phosphatic fertilizer type affects the accumulation of cadmium (Cd) and fluorine (F) in pasture herbage. North Carolina phosphate rock and partially acidulated fertilizers derived from this rock generally have higher Cd and F concentrations compared to single superphosphate currently manufactured in Australia. Clover herbage from sites of the National Reactive Phosphate Rock (RPR) trial was collected and analysed for concentrations of Cd (11 sites) and F (4 sites). A comparison was made between pastures fertilized with 4 rates of single superphosphate, North Carolina phosphate rock, and partially acidulated phosphate rock having Cd concentrations of 283, 481, and 420 mg Cd/kg P respectively, and 170, 271, and 274 g F/kg P respectively. One site used Hemrawein (Egypt) phosphate rock (HRP) having a Cd and F concentration of 78 mg Cd/kg P and 256 g F/kg P respectively. To help identify differences in herbage Cd concentrations between sites, unfertilised soils from each site were analyzed for total and extractable Cd contents. At one site Cd concentrations in bulk herbage (clover, grasses and weeds) were related to infestation of the pasture by capeweed (Arctotheca calendula L. Levyns). There were no significant differences between F in herbage from plots fertilized with single superphosphate, partially acidulated phosphate rock or North Carolina phosphate rock, or between sites. Concentrations of F in herbage were low, generally less than 10 mg F /kg. However, there were large differences in Cd concentrations in herbage between sites, while differences between fertilizer treatments were small in comparison. The site differences were only weakly related to total or extractable (0.01 mol/L CaCl 2 ) Cd concentrations in soil. Significant differences in Cd concentrations in clover due to fertilizer type were found at 5 sites. North Carolina phosphate rock treatments had significantly higher Cd concentrations in clover compared to

  16. Assessment of Protective Effect of Some Modern Agrochemicals against Ozone-Induced Stress in Sensitive Clover and Tobacco Cultivars

    OpenAIRE

    Blum, Oleg; Didyk, Nataliya; Pavluchenko, Nataliya; Godzik, Barbara

    2011-01-01

    Some modern agrochemicals with antioxidant potential were tested for their protective effect against ozone injury using clover and tobacco ozone-sensitive cultivars as model plants subjected to ambient ozone at two sites (Kyiv city in Ukraine and Szarów village in Poland). All used agrochemicals showed partial protective effects against ozone injury on clover and tobacco. Conducted studies confirmed the effectiveness of modern fungicides belonging to strobilurin group as protectants of sensit...

  17. Determination of toxicity limiting values of Zn, Cu, and Pb for oat and red clover

    Energy Technology Data Exchange (ETDEWEB)

    Hodenberg, A V; Finck, A

    1975-01-01

    Toxicity limiting values of Zn, Cu and Pb are determined in order to investigate the causes of growth damages in certain fields. Since the true toxicity limit is difficult to estimate, a somewhat higher content is called the toxicity limiting value resulting after the subtraction of a significant yield difference. The pot experiments with increasing fertilization of Zn, Cu and Pb in soil cultures gave the following results. For Zn, the toxicity limit is 410 ppm in oats at the beginning of tilling. In red clover six weeks of age, it is only 290 ppm of Zn and therefore much lower. For Cu, the toxicity limit is 20 ppm in oats at the beginning of tilling and 18 ppm in six weeks old red clover. For Pb, a toxic effect could be observed above 50 ppm, but this does not seem to be the true limit because of disturbing salt effects.

  18. Nitrogen transfer via VA mycorrhiza hyphae link between roots of red clover

    International Nuclear Information System (INIS)

    Zhang Junling; Li Xiaolin; Yang Zhifu

    1997-01-01

    Five compartments with a air gap was designed to examine underground N transfer via VAM hyphae link between roots of red clover. 15 N was injected into the root compartment for donor plant 4 weeks after planting. Plants were harvested 6 weeks afterwards. The experiment had four treatments: two N levels (receiver root compartment); mycorrhiza and non-mycorrhiza treatment (B compartment), and each with four replicates. It was demonstrated that there was no significant difference in 15 N abundance in receiver red clover regardless of the inoculation of mycorrhiza fungi and nitrogen fertilization. Both shoot and root dry weight of receiver plants were not affected too, however, plant P and N nutrient status got better under this condition

  19. Modelling Gene Flow between Fields of White Clover with Honeybees as Pollen Vectors

    DEFF Research Database (Denmark)

    Løjtnant, Christina; Boelt, Birte; Clausen, Sabine Karin

    2012-01-01

    The portion-dilution model is a parametric restatement of the conventional view of animal pollination; it predicts the level of pollinator-mediated gene dispersal. In this study, the model was applied to white clover (Trifolium repens) and its most frequent pollinator, the honeybee (Apis mellifera......). One of the three parameters in the portion-dilution model is the mean number of flowers a pollinator visits in one foraging bout. An alternative method to estimate this parameter was developed that was not depending on pollinator hive-seeking behaviour. The new estimation method, based on nectar......% with an estimated 95% percentile of 70%. The results show that the European Union threshold limit of 0.9% GM admixture for food and feed will likely be exceeded at times and especially organic farmers that do not accept GM admixture and often have clover and clover–grass fields might face challenges with admixture...

  20. Crystallization and preliminary X-ray diffraction analysis of red clover necrotic mosaic virus

    International Nuclear Information System (INIS)

    Martin, Stanton L.; Guenther, Richard H.; Sit, Tim L.; Swartz, Paul D.; Meilleur, Flora; Lommel, Steven A.; Rose, Robert B.

    2010-01-01

    Virions of red clover necrotic mosaic virus have been purified and crystallized. The space group was determined to be I23, with unit-cell parameter a = 377.8 Å. The crystals diffracted to 4 Å resolution. Red clover necrotic mosaic virus (RCNMV) is a species that belongs to the Tombusviridae family of plant viruses with a T = 3 icosahedral capsid. RCNMV virions were purified and were crystallized for X-ray analysis using the hanging-drop vapor-diffusion method. Self-rotation functions and systematic absences identified the space group as I23, with two virions in the unit cell. The crystals diffracted to better than 4 Å resolution but were very radiation-sensitive, causing rapid decay of the high-resolution reflections. The data were processed to 6 Å in the analysis presented here

  1. Performance comparison for modes of operation and suppression cases of the clover detector

    International Nuclear Information System (INIS)

    Kshetri, R

    2014-01-01

    Peak-to-total ratios have been extracted for a segmented clover detector for various modes of operation and suppression cases. Based on absorption and scattering of gamma-rays, a formalism has been presented for quantifying the advantages of using addback mode and active suppression. Further investigations have been performed to find if the reduction of background counts is caused more by active suppression or by addback process

  2. Effects of oil sands effluent on cattail and clover: photosynthesis and the level of stress proteins

    Energy Technology Data Exchange (ETDEWEB)

    Crowe, A.U.; Han, B.; Kermode, A.R.; Bendell-Young, L.I.; Plant, A.L. [Simon Fraser University, Burnaby (Canada). Dept. of Biological Sciences

    2001-07-01

    The oil sands industry located in northeastern Alberta, Canada, generates large volumes of effluent characterized by a high level of dissolved ions and naphthenic acids. The dikes used to store the effluent seep, creating wetlands which are subsequently invaded by obligate wetland flora such as cattail (Typha latifolia L.). The appearance of these wetlands prompted the oil sands industry to consider wetlands as part of their reclamation strategy. However, to ensure long-term viability of such wetlands, the response of the flora to the industrial effluent needed to be determined. To this end, apparent photosynthesis (APS), the level of ribulose-1,5-bisphosphate carboxylase (RuBisCo) large subunit, dehydrin-related polypeptides, and protein disulphide isomerase (PDI) were evaluated in cattail and alsike clover plants (Trifolium hybridum L.) exposed to the oil sands effluent. APS measured in plants impacted by oil sands effluent was significantly higher than that of plants in the non-impacted off-site location. Among the on-site locations, plants growing in the natural wetlands site had higher APS compared to all other sites. The level of RuBisCo was not increased in cattail or clover growing in effluent-contaminated sites indicating that enhanced photosynthesis was not due to greater levels of this enzyme. Dehydrin-related polypeptides were detected only in the roots of cattail and were absent in clover. The polypeptide profile was altered in cattail exposed to oil sands effluent indicating that they were responding to an osmotic stress. The level of PDI was unaffected in the leaves of cattail regardless of the nature of the effluent to which they were exposed. Overall, the data indicate that cattail and clover are adapted to the oil sands effluent, although further studies are needed to assess their long-term ability to survive in the presence of this anthropogenic stress. (Author)

  3. Inhibitory potential of pure isoflavonoids, red clover, and alfalfa extracts on hemoglobin glycosylation

    Directory of Open Access Journals (Sweden)

    Mohsen Hosseini

    2015-03-01

    Full Text Available BACKGROUND: Non-enzymatic glycosylation of hemoglobin is complications of diabetes. Antioxidant system imbalance can result in the emergence of free radicals’ destructive effects in the long-term. Red clover (Trifolium pratense L. and alfalfa (Medicago sativa L. contain isoflavonoids and have antioxidant activity. This experimental study evaluated the inhibitory activity of pure isoflavonoids (daidzein and genistein, red clover and alfalfa extracts on hemoglobin glycosylation. METHODS: This study was performed in Iran. Stock solution of hydroalcoholic extracts of red clover and alfalfa in concentrations of 1 and 10 g/100 ml and stock solution of daidzein and genistein in concentrations of 250 ng, 500 ng, 25 µg and 250 µg/100 ml were prepared as case groups. Control group was without hydroalcoholic extracts of plants and pure isoflavonoids. All experiments were performed in triplicate. Hemoglobin was prepared and antioxidant activities were investigated to estimate degree of nonenzymatic hemoglobin glycosylation. RESULTS: There was no significantly difference between used extracts (extract of red clover and alfalfa and control of the hemoglobin glycosylation but using daidzein (P = 0.046, 0.029 and 0.021, respectively and genistein (P = 0.034, 0.036 and 0.028 significantly inhibited (P < 0.050 this reaction in 25 µg/100 ml, 250 and 500 ng/100 ml concentrations when compared to control. in 25 µg/100 ml, 250 ng and 500 ng/100 ml concentrations percentage of inhibition were 32, 80 and 74.5% respectively with used of daidzein and were 21, 83 and 76% respectively with consumption of genistein. CONCLUSION: According to decrease of glycation of hemoglobin with isoflavonoids, two used plant in this study containing isoflavonoid may be useful on diabetes.   

  4. Cool-season annual pastures with clovers to supplement wintering beef cows nursing calves

    Directory of Open Access Journals (Sweden)

    Gunter Stacey A

    2012-07-01

    Full Text Available Abstract In December of 3 years, 87 beef cows with nursing calves (594 ± 9.8 kg; calving season, September to November at side were stratified by body condition score, body weight, cow age, and calf gender and divided randomly into 6 groups assigned to 1 of 6 cool-season annual pastures (0.45 ha/cow that had been interseeded into a dormant common bermudagrass (Cynodon dactylon [L.] Pers./bahiagrass (Paspalum notatum Flugge sod. Pastures contained 1 of the following 3 seeding mixtures (2 pastures/mixture: 1 wheat (Triticum aestivum L. and ryegrass (Lolium multiflorum Lam., WRG, 2 wheat and ryegrass plus red clover (Trifolium pretense L., WRR, or 3 wheat and ryegrass plus white (Trifolium repens L. and crimson clovers (Trifolium incarnatum L., WRW. All groups had ad libitum access to grass hay (12% crude protein; 58% total digestible nutrients. The second week in December, cow estrous cycles were synchronized and artificially inseminated. In late December, a bull was placed with each group for 60-d. Data were analyzed with an analysis of variance using a mixed model containing treatment as the fixed effect and year as the random effect. Body weight and condition scores did not differ (P ≥ 0.27 among cows between February and June. Calf birth weights or average daily gain did not differ (P ≥ 0.17 among treatments; however, calves grazing pastures with clovers did tend (P = 0.06 to weigh more than calves grazing grass only. Weaning weight per cow exposed to a bull was greater (P = 0.02 for WRR and WRW than WRG. Cows grazing winter-annual pastures containing clovers tended to wean more calf body weight per cow exposed to a bull than cows grazing the grass only pastures.

  5. Investigation of Amino Acids As Herbicides for Control of Orobanche minor Parasitism in Red Clover

    Directory of Open Access Journals (Sweden)

    Mónica Fernández-Aparicio

    2017-05-01

    Full Text Available Certain amino acids induce inhibitory effects in plant growth due to feedback inhibition of metabolic pathways. The inhibition patterns depend on plant species and the plant developmental stage. Those amino acids with inhibitory action on specific weeds could be utilized as herbicides, however, their use for weed control has not been put into practice. Orobanche minor is a weed that parasitizes red clover. O. minor germination is stimulated by clover root exudates. The subsequent seedling is an obligated parasite that must attach quickly to the clover root to withdraw its nutrients. Early development of O. minor is vulnerable to amino acid inhibition and therefore, a series of in vitro, rhizotron, and field experiments were conducted to investigate the potential of amino acids to inhibit O. minor parasitism. In in vitro experiments it was found that among a collection of 20 protein amino acids, lysine, methionine and tryptophan strongly interfere with O. minor early development. Field research confirmed their inhibitory effect but revealed that methionine was more effective than lysine and tryptophan, and that two successive methionine applications at 308 and 543 growing degree days inhibited O. minor emergence in red clover up to 67%. We investigated additional effects with potential to influence the practical use of amino acids against broomrape weeds, whether the herbicidal effect may be reversible by other amino acids exuded by host plants or may be amplified by inducing host resistance barriers against O. minor penetration. This paper suggests that amino acids may have the potential to be integrated into biorational programs of broomrape management.

  6. Investigation of Amino Acids As Herbicides for Control of Orobanche minor Parasitism in Red Clover.

    Science.gov (United States)

    Fernández-Aparicio, Mónica; Bernard, Alexandre; Falchetto, Laurent; Marget, Pascal; Chauvel, Bruno; Steinberg, Christian; Morris, Cindy E; Gibot-Leclerc, Stephanie; Boari, Angela; Vurro, Maurizio; Bohan, David A; Sands, David C; Reboud, Xavier

    2017-01-01

    Certain amino acids induce inhibitory effects in plant growth due to feedback inhibition of metabolic pathways. The inhibition patterns depend on plant species and the plant developmental stage. Those amino acids with inhibitory action on specific weeds could be utilized as herbicides, however, their use for weed control has not been put into practice. Orobanche minor is a weed that parasitizes red clover. O. minor germination is stimulated by clover root exudates. The subsequent seedling is an obligated parasite that must attach quickly to the clover root to withdraw its nutrients. Early development of O. minor is vulnerable to amino acid inhibition and therefore, a series of in vitro , rhizotron, and field experiments were conducted to investigate the potential of amino acids to inhibit O. minor parasitism. In in vitro experiments it was found that among a collection of 20 protein amino acids, lysine, methionine and tryptophan strongly interfere with O. minor early development. Field research confirmed their inhibitory effect but revealed that methionine was more effective than lysine and tryptophan, and that two successive methionine applications at 308 and 543 growing degree days inhibited O. minor emergence in red clover up to 67%. We investigated additional effects with potential to influence the practical use of amino acids against broomrape weeds, whether the herbicidal effect may be reversible by other amino acids exuded by host plants or may be amplified by inducing host resistance barriers against O. minor penetration. This paper suggests that amino acids may have the potential to be integrated into biorational programs of broomrape management.

  7. Cleaning the Soil from Zinc Using Red Clovers “Arimaičiai”

    OpenAIRE

    Audronė Mikalajūnė; Giedrė Jasulaitytė

    2011-01-01

    Zinc as a nutrition element is required to plants in small quantities to maintain normal functions of metabolism mechanisms. Our work analyses the efficiency of red clovers “Arimaičiai” for cleaning zinc from the soil contaminated with zinc under laboratory conditions. Seeds were sown in three differently polluted soils: clean soil, once contaminated with zinc and periodically contaminated with zinc soil. Zinc concentration in one time contaminated soil was 45 mg/kg. After 6 months of phytore...

  8. Effects of Titanium Dioxide Nanoparticles on Red Clover and Its Rhizobial Symbiont.

    Science.gov (United States)

    Moll, Janine; Okupnik, Annette; Gogos, Alexander; Knauer, Katja; Bucheli, Thomas D; van der Heijden, Marcel G A; Widmer, Franco

    2016-01-01

    Titanium dioxide nanoparticles (TiO2 NPs) are in consideration to be used in plant protection products. Before these products can be placed on the market, ecotoxicological tests have to be performed. In this study, the nitrogen fixing bacterium Rhizobium trifolii and red clover were exposed to two TiO2 NPs, i.e., P25, E171 and a non-nanomaterial TiO2. Growth of both organisms individually and their symbiotic root nodulation were investigated in liquid and hydroponic systems. While 23 and 18 mg l-1 of E171 and non-nanomaterial TiO2 decreased the growth rate of R. trifolii by 43 and 23% respectively, P25 did not cause effects. Shoot length of red clover decreased between 41 and 62% for all tested TiO2 NPs. In 21% of the TiO2 NP treated plants, no nodules were found. At high concentrations certain TiO2 NPs impaired R. trifolii as well as red clover growth and their symbiosis in the hydroponic systems.

  9. Effects of Titanium Dioxide Nanoparticles on Red Clover and Its Rhizobial Symbiont.

    Directory of Open Access Journals (Sweden)

    Janine Moll

    Full Text Available Titanium dioxide nanoparticles (TiO2 NPs are in consideration to be used in plant protection products. Before these products can be placed on the market, ecotoxicological tests have to be performed. In this study, the nitrogen fixing bacterium Rhizobium trifolii and red clover were exposed to two TiO2 NPs, i.e., P25, E171 and a non-nanomaterial TiO2. Growth of both organisms individually and their symbiotic root nodulation were investigated in liquid and hydroponic systems. While 23 and 18 mg l-1 of E171 and non-nanomaterial TiO2 decreased the growth rate of R. trifolii by 43 and 23% respectively, P25 did not cause effects. Shoot length of red clover decreased between 41 and 62% for all tested TiO2 NPs. In 21% of the TiO2 NP treated plants, no nodules were found. At high concentrations certain TiO2 NPs impaired R. trifolii as well as red clover growth and their symbiosis in the hydroponic systems.

  10. PERSISTENCE ASSESSMENT OF RED CLOVER (Trifolium pratense L. IN TÂRGOVISTE PLAIN

    Directory of Open Access Journals (Sweden)

    D. DUNEA

    2008-05-01

    Full Text Available Since the most important deficiency of forage stands is the inability to maintain adequate legume participation in mixture, it is the purpose of this paper to examine persistence in red clover in Târgoviste Plain eco-climatic conditions, together with the factors that affect it. Six red clover cultivars (Napoca-Tetra, Dacia Tetra, Vesna – tetraploids; Flora, Roxana, Start – diploids and one white clover diploid cultivar (Karina were used in pure culture and in mixture (50:50 with hybrid ryegrass (Zefir – tetraploid in a randomized block design with three replicates. Ground cover assessment in early spring was a suggestive indicator of the stand persistence to define the stability and sustainability boundaries of a reliable intensive system. In the beginning of the third year of cropping, ground cover was 54.33% for tetraploid cultivars (CV = 43.25%, and 67% for diploid cultivars (CV = 6.83% in pure stands. Same ground cover average of 27% was established both for tetraploid cultivars (CV = 36.47%, and for diploid cultivars (CV = 16.97% in mixtures with hybrid ryegrass.

  11. The "clover technique" as a novel approach for correction of post-traumatic tricuspid regurgitation.

    Science.gov (United States)

    Alfieri, O; De Bonis, M; Lapenna, E; Agricola, E; Quarti, A; Maisano, F

    2003-07-01

    To describe a novel technique, named "clover," to correct complex post-traumatic tricuspid valve lesions. Five patients with severe post-traumatic tricuspid insufficiency underwent valve reconstruction with the clover technique, a new surgical approach that consists of stitching together the middle point of the free edges of the tricuspid leaflets, producing a clover-shaped valve. The mechanism of tricuspid regurgitation was complex in all patients, and right ventricular function was always moderately to severely depressed. An echocardiographic study was performed after cardiopulmonary bypass, at discharge, and at follow-up. Cardiopulmonary bypass time was 32 +/- 6.3 minutes and crossclamp time was 23 +/- 7.4. There was no hospital mortality or morbidity. Intraoperative transesophageal and predischarge transthoracic echocardiography showed perfect results in all patients. No late deaths occurred. At the latest follow-up, extending to 14.2 months (mean 11.3; median 12.4), all patients were asymptomatic (New York Heart Association class I) with trivial (2 patients) or no residual regurgitation (3 patients) on 2-dimensional echocardiogram. No transvalvular gradient was revealed in any patient. A significant reduction of the right ventricular end-diastolic dimensions was noted as well (from 54 +/- 7.1 mm to 40 +/- 7.5 mm, P tricuspid valve repair in case of severe traumatic tricuspid valve insufficiency, leading to very satisfactory mid-term results even in the presence of complex lesions or dilatation and deterioration of the right ventricle.

  12. Cleaning the Soil from Zinc Using Red Clovers “Arimaičiai”

    Directory of Open Access Journals (Sweden)

    Audronė Mikalajūnė

    2011-02-01

    Full Text Available Zinc as a nutrition element is required to plants in small quantities to maintain normal functions of metabolism mechanisms. Our work analyses the efficiency of red clovers “Arimaičiai” for cleaning zinc from the soil contaminated with zinc under laboratory conditions. Seeds were sown in three differently polluted soils: clean soil, once contaminated with zinc and periodically contaminated with zinc soil. Zinc concentration in one time contaminated soil was 45 mg/kg. After 6 months of phytoremediation, the remained zinc concentration in the soil was 3 times lower comparing with the initial concentration. It was also determined that under such conditions, the uptake of red clovers made approximately 65% of zinc. Permanent soil contamination with zinc increased concentration before phytoremediation up to 80 mg/kg. After 6 months of phytoremediation, zinc concentration was determined to be 1.9 times lower. Otherwise, the soil was permanently contaminated with larger zinc quantities and after application of which reached 300 mg/kg. In this case, following half a year of phytoremediation, zinc concentration in the soil was 1.7 times lower comparing with the initial concentration after contamination. It was determined that the uptake of red clovers made approximately 17% of zinc.Article in Lithuanian

  13. Effects of grass-clover management and cover crops on nitrogen cycling and nitrous oxide emissions in a stockless organic crop rotation

    DEFF Research Database (Denmark)

    Brozyna, Michal Adam; Petersen, Søren O; Chirinda, Ngoni

    2013-01-01

    Nitrogen (N) supply in stockless organic farming may be improved through use of grass-clover for anaerobic digestion, producing biogas and digested manure for use as fertilizer in the crop rotation. We studied the effects of grass-clover management on N cycling, nitrous oxide (N2O) emissions...... in the rotation (spring barley, potato and winter wheat); actual digestion of grass-clover cuttings was not possible, instead digested pig manure was used as substitute for digested grass-clover. Nitrous oxide fluxes were monitored between April 2008 and May 2009. In general, application of digested manure had...

  14. Production of N{sub 2}O in grass-clover pastures

    Energy Technology Data Exchange (ETDEWEB)

    Carter, M.S.

    2005-09-01

    Agricultural soils are known to be a considerable source of the strong greenhouse gas nitrous oxide (N{sub 2}O), and in soil N{sub 2}O is mainly produced by nitrifying and denitrifying bacteria. In Denmark, grass-clover pastures are an important component of the cropping system in organic as well as conventional dairy farming, and on a European scale grass-clover mixtures represent a large part of the grazed grasslands. Biological dinitrogen (N{sub 2}) fixation in clover provides a major N input to these systems, but knowledge is sparse regarding the amount of fixed N{sub 2} lost from the grasslands as N2O. Furthermore, urine patches deposited by grazing cattle are known to be hot-spots of N{sub 2}O emission, but the mechanisms involved in the N{sub 2}O production in urine-affected soil are very complex and not well understood. The aim of this Ph.D. project was to increase the knowledge of the biological and physical-chemical mechanisms, which control the production of N2O in grazed grass-clover pastures. Three experimental studies were conducted with the objectives of: 1: assessing the contribution of recently fixed N{sub 2} as a source of N{sub 2}O. 2: examining the link between N{sub 2}O emission and carbon mineralization in urine patches. 3: investigating the effect of urine on the rates and N{sub 2}O loss ratios of nitrification and denitrification, and evaluating the impact of the chemical conditions that arise in urine affected soil. The results revealed that only 3.2 {+-} 0.5 ppm of the recently fixed N{sub 2} was emitted as N2O on a daily basis. Thus, recently fixed N released via easily degradable clover residues appears to be a minor source of N2O. Furthermore, increased N{sub 2}O emission following urine application at rates up to 5.5 g N m{sup -2} was not caused by enhanced denitrification stimulated by labile compounds released from scorched plant roots. Finally, the increase of soil pH and ammonium following urine application led to raised

  15. Joseph Clover and the cobra: a tale of snake envenomation and attempted resuscitation with bellows in London, 1852.

    Science.gov (United States)

    Ball, C

    2010-07-01

    The Industrial Revolution saw the creation of many new jobs, but probably none more curious than that of zookeeper. The London Zoological Gardens, established for members in 1828, was opened to the general public in 1847. In 1852 the "Head Keeper in the Serpent Room", Edward Horatio Girling, spent a night farewelling a friend departing for Australia. He arrived at work in an inebriated state and was bitten on the face by a cobra that he was handling in a less than sensible manner. He was taken by cab to University College Hospital where he was resuscitated by a number of doctors, including Joseph Clover then the resident medical officer to the hospital and later to become the leading anaesthetist in London. Clover recorded this event in his diary along with the resuscitation method used. The patient eventually died but his treatment created a flurry of correspondence in the medical and lay press. Interestingly, the attempted resuscitation was with bellows, which had been abandoned by the Royal Humane Society twenty years earlier Clover records other cases of resuscitation with bellows at University College Hospital during his time as a resident medical officer there (1848 to 1853). There is a casebook belonging to Joseph Clover in the Geoffrey Kaye Museum, in Melbourne. This story is one of the many interesting stories uncovered during a study of this book and Clover's other personal papers.

  16. Homogentisate solanesyl transferase (HST) cDNA’s in maize

    Science.gov (United States)

    Maize white seedling 3 (w3) has served as a model albino-seedling mutant since its discovery in 1923. We show that the w3 phenotype is caused by disruptions in homogentisate solanesyl transferase (HST), an enzyme that catalyzes the committed step in plastoquinone-9 (PQ9) biosynthesis. This reaction ...

  17. Insecticide resistance and glutathione S-transferases in mosquitoes ...

    African Journals Online (AJOL)

    Mosquito glutathione S-transferases (GSTs) have received considerable attention in the last 20 years because of their role in insecticide metabolism producing resistance. Many different compounds, including toxic xenobiotics and reactive products of intracellular processes such as lipid peroxidation, act as GST substrates.

  18. Steroid sulfatase and sulfuryl transferase activities in human brain tumors

    Czech Academy of Sciences Publication Activity Database

    Kříž, L.; Bičíková, M.; Mohapl, M.; Hill, M.; Černý, Ivan; Hampl, R.

    2008-01-01

    Roč. 109, č. 1 (2008), s. 31-39 ISSN 0960-0760 Institutional research plan: CEZ:AV0Z40550506 Keywords : dehydroepiandrosterone * steroid sulfatase * steroid sulfuryl transferase * brain Subject RIV: CC - Organic Chemistry Impact factor: 2.827, year: 2008

  19. The performance of a white clover-based dairy system in comparison with a grass/fertiliser-N system. II. Animal production, economics and environment

    NARCIS (Netherlands)

    Schils, R.L.M.; Boxem, T.; Jagtenberg, C.J.; Verboom, M.C.

    2000-01-01

    The performance of a white clover based dairy system in comparison with a grass/fertiliser-N system was studied during three years. Both systems had 59 cows, plus young stock, on an area of 40.6 ha for grass/clover and 34.4 ha for grass/fertiliser-N. During the grazing season, the cows in both

  20. 21 CFR 862.1030 - Alanine amino transferase (ALT/SGPT) test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Alanine amino transferase (ALT/SGPT) test system... Test Systems § 862.1030 Alanine amino transferase (ALT/SGPT) test system. (a) Identification. An alanine amino transferase (ALT/SGPT) test system is a device intended to measure the activity of the...

  1. Biological activity of clovers - free radical scavenging ability and antioxidant action of six Trifolium species.

    Science.gov (United States)

    Kolodziejczyk-Czepas, Joanna; Nowak, Pawel; Kowalska, Iwona; Stochmal, Anna

    2014-10-01

    Clovers were chosen on the basis of traditional medicine recommendations, agricultural value, or available information on their promising chemical profiles. This study evaluates and compares free radical scavenging and antioxidant properties of six clover species: Trifolium alexandrinum L. (Leguminosae), Trifolium fragiferum L., Trifolium hybridum L., Trifolium incarnatum L., Trifolium resupinatum var. majus Boiss., and Trifolium resupinatum var. resupinatum L. Free radical scavenging activity of the extracts (1.5-50 µg/ml) was estimated by reduction of 1,1-diphenyl-2-picrylhydrazyl (DPPH(•)) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic) acid (ABTS(•)) radicals. The Trifolium extract effects on total antioxidant capacity of blood plasma were determined by the reduction of ABTS(•+) and DPPH(•) radicals, as well as with the use of the ferric reducing ability of plasma (FRAP) assay. The UPLC analysis of chemical profiles of the examined extracts showed the presence of three or four groups of phenolic substances, including phenolic acids, clovamides, isoflavones, and other flavonoids. The measurements of free radical scavenging and ferric reducing ability of the examined clover extracts revealed the strongest effect for T. alexandrinum. Furthermore, antioxidant activity assays in human plasma have shown protective effects of all extracts against peroxynitrite-induced reduction of total antioxidant capacity. Trifolium plants may be a rich source of bioactive substances with antioxidant properties. The examined extracts displayed free radical scavenging action and partly protected blood plasma against peroxynitrite-induced oxidative stress; however, the beneficial effects of T. alexandrinum and T. incarnatum seem to be slightly higher.

  2. Effect of aditive supplementation to ensiled red clover on voluntary intake, digestibility and N balance

    Directory of Open Access Journals (Sweden)

    Marina Vranić

    2009-06-01

    Full Text Available The objective of this study was to determine effect of additive supplementation to red clover silage on ad libitum intake of fresh silage and dry matter (DM, in vivo digestibility of DM, organic matter (OM, OM in DM (D-value, crude protein (CP and nitrogen (N balance. Red clover was harvested at 60% bloom stage. It was ensiled into round bales without an additive (CD and with an additive supplementation (CDA in the amount of 2 L t-1 fresh material. Statistically lower (P<0.001 DM content was recorded in CD (405 g kg-1 fresh sample in comparison with CDA (665 g ST kg-1 fresh sample. Statistically higher CP content (P<0.001 was recorded in CD (127 g kg-1 ST in comparison with CDA (110 g SP kg-1 ST. CD had lower pH (P<0.001 (4.9 in comparison with CDA (5.2. No differences were recorded in NH3 between treatments. Ad libitum intake of fresh silage and silage DM was higher (P<0.001 and P<0.01, respectively in CD in comparison with CDA. Digestibility of DM, OM, CP, D-value and N balance were not different between treatments. It was concluded that the positive effect of additive supplementation to red clover silage on chemical composition, ad libitum intake, digestibility and N balance was not recorded due to applied ensiling technology as additive can improve feeding value of roughage, but it is not a replacement for good management practices.

  3. Characterisation of a Compton suppressed Clover detector for high energy gamma rays (=<11MeV)

    International Nuclear Information System (INIS)

    Saha Sarkar, M.; Kshetri, Ritesh; Raut, Rajarshi; Mukherjee, A.; Sinha, Mandira; Ray, Maitreyi; Goswami, A.; Roy, Subinit; Basu, P.; Majumder, H.; Bhattacharya, S.; Dasmahapatra, B.

    2006-01-01

    Gamma ray spectra of two (p,γ) resonances have been utilised for the characterisation of the Clover detector at energies beyond 5MeV. Apart from the efficiency and the resolution of the detector, the shapes of the full energy peaks as well as the nature of the escape peaks which are also very crucial at higher energies have been analysed with special attention. Proper gain matching in software have checked deterioration in the energy resolution and distortion in the peak shape due to addback. The addback factors show sharp increasing trend even at energies around 11MeV

  4. Characterisation of a Compton suppressed Clover detector for high energy gamma rays (=<11MeV)

    Energy Technology Data Exchange (ETDEWEB)

    Saha Sarkar, M. [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India)]. E-mail: maitrayee.sahasarkar@saha.ac.in; Kshetri, Ritesh [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Raut, Rajarshi [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Mukherjee, A. [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Sinha, Mandira [Gurudas College, Narkeldanga, Kolkata-700054 (India); Ray, Maitreyi [Behala College, Parnashree, Kolkata-700060 (India); Goswami, A. [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Roy, Subinit [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Basu, P. [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Majumder, H. [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Bhattacharya, S. [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India); Dasmahapatra, B. [Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064 (India)

    2006-01-01

    Gamma ray spectra of two (p,{gamma}) resonances have been utilised for the characterisation of the Clover detector at energies beyond 5MeV. Apart from the efficiency and the resolution of the detector, the shapes of the full energy peaks as well as the nature of the escape peaks which are also very crucial at higher energies have been analysed with special attention. Proper gain matching in software have checked deterioration in the energy resolution and distortion in the peak shape due to addback. The addback factors show sharp increasing trend even at energies around 11MeV.

  5. The influence of phosphorus deficiency on growth and nitrogen fixation of white clover plants

    OpenAIRE

    Høgh-Jensen, Dr Henning; Schjoerring, Dr Jan K.; Soussana, Dr Jean-Francois

    2002-01-01

    The effects of P deficiency on growth, N2‐fixation and photosynthesis in white clover (Trifolium repens L.) plants were investigated using three contrasting relative addition rates of P, or following abrupt withdrawal of the P supply. Responses to a constant below‐optimum P supply rate consisted of a decline in N2‐fixation per unit root weight and a small reduction in the efficiency with which electrons were allocated to the reduction of N2 in nodules. Abrupt removal of P arrested nodule grow...

  6. Nucleon form factors and structure functions from Nf=2 Clover fermions

    International Nuclear Information System (INIS)

    Collins, S.; Goeckeler, M.; Haegler, P.

    2010-12-01

    We give an update on our ongoing efforts to compute the nucleon's form factors and moments of structure functions using N f =2 flavours of non-perturbatively improved Clover fermions. We focus on new results obtained on gauge configurations where the pseudo-scalar meson mass is in the range of 170-270 MeV. We compare our results with various estimates obtained from chiral effective theories since we have some overlap with the quark mass region where results from such theories are believed to be applicable. (orig.)

  7. Short-term residual N unaffected by forbs in grass-clover mixtures

    DEFF Research Database (Denmark)

    Dhamala, Nawa Raj; Rasmussen, Jim; Cong, Wenfeng

    2017-01-01

    We determined the effect on residual nitrogen (N) of including forbs (chicory, ribwort plantain and caraway) in perennial ryegrass-red clover mixtures. Although soil N inputs during the grassland phase differed markedly between mixtures, in a pot experiment we found no differences...... in the potentially mineralizable N of the soil or in the dry matter production and N content of the spring barley test crop. The fertilizer value of the grassland mixtures corresponded to 10 g N/m2, irrespective of forb inclusion. Thus, the inclusion of nonlegume forbs did not negatively affect short-term residual N...

  8. Effects of field characteristics on abundance of bumblebees (Bombus spp.) and seed yield in red clover fields

    DEFF Research Database (Denmark)

    Wermuth, Kirsten Haugaard; Dupont, Yoko L.

    2010-01-01

    Red clover is a key floral ressource for bumblebees (Bombus spp.).We here investigate variation within and among red clover fields in species richness and abundance of Bombus spp. in addition to Apis mellifera. Bumblebee individuals were grouped into the following functional groups, based on castes...... and tongue length: (1) all queens, (2) all workers, (3) short-tongued workers and (4) long-tongued workers. In 14 study fields, no spatial or diurnal within-field differences were found in abundances of bee groups. However, seasonal differences were detected. On average 6.3±0.6 Bombus spp. were observed...

  9. Assessment of dietary ratios of red clover and corn silages on milk production and milk quality in dairy cows.

    Science.gov (United States)

    Moorby, J M; Ellis, N M; Davies, D R

    2016-10-01

    Twenty-four multiparous Holstein-Friesian dairy cows were used in a replicated 3×3 Latin square changeover design experiment to test the effects of changing from corn (Zea mays) silage to red clover (Trifolium pratense) silage in graded proportions on feed intakes, milk production, and whole-body N and P partitioning. Three dietary treatments with ad libitum access to 1 of 3 forage mixtures plus a standard allowance of 4kg/d dairy concentrates were offered. The 3 treatment forage mixtures were, on a dry matter (DM) basis: (1) R10: 90% corn silage and 10% red clover silage, (2) R50: 50% corn silage and 50% red clover silage, and (3) R90: 10% corn silage and 90% red clover silage. In each of 3 experimental periods, there were 21d for adaptation to diets, and 7d for measurements. Diet crude protein intakes increased, and starch intakes decreased, as the silage mixture changed from 90% corn to 90% red clover, although the highest forage DM intakes and milk yields were achieved on diet R50. Although milk fat yields were unaffected by diet, milk protein yields were highest with the R 0250 diet. Whole-body partitioning of N was measured in a subset of cows (n=9), and both the daily amount and proportion of N consumed that was excreted in feces and urine increased as the proportion of red clover silage in the diet increased. However, the apparent efficiency of utilization of feed N for milk protein production decreased from 0.33g/g for diet R10 to 0.25g/g for diet R90. The urinary excretion of purine derivatives (sum of allantoin and uric acid) tended to increase, suggesting greater flow of microbial protein from the rumen, as the proportion of red clover silage in the diet increased, and urinary creatinine excretion was affected by diet. Fecal shedding of E. coli was not affected by dietary treatment. In conclusion, even though microbial protein flow may have been greatest from the R 0450 diet, optimum feed intakes and milk yields were achieved on a diet that contained a

  10. Effect of microgravity on stress ethylene and carbon dioxide production in sweet clover (Melilotus alba L.)

    Science.gov (United States)

    Gallegos, Gregory L.; Odom, William R.; Guikema, James A.

    1995-01-01

    The study of higher plant growth and development in the microgravity (micro-g) environment continues to be a challenge. This is in part a result of the available flight qualified hardware with restrictive closed gas environments. This point is underscored by considering that gas exchange of seedlings grown in microgravity may be further limited owing to a thicker layer of water wicked onto the roots and to the absence of convective mixing. We hypothesized that seedlings grown under such conditions will experience greater hypoxia in microgravity than at Earth gravity, and thus produce greater stress ethylene. We compared flight and ground samples of sweet clover seedlings grown in the Fluid Processing Apparatus (FPA) during STS-57 and found them to contain extremely high levels of carbon dioxide (CO2) and stress ethylene. There were time dependent increases for both gases, and seedling growth was greatly inhibited. We repeated these experiments aboard STS-60 using modified chambers which increased, by fifty fold, the air available to the developing seedlings. Sweet clover seed germination and subsequent seedling growth to eight days within the FPA modified with a gas permeable membrane is not compromised by the microgravity environment.

  11. NUTRITIVE VALUES OF SOME ANNUAL CLOVERS (Trifolium sp. AT DIFFERENT GROWTH STAGES

    Directory of Open Access Journals (Sweden)

    A TEKELI

    2006-05-01

    Full Text Available This study was conducted between the years of 2001-2002 in the experimental area and laboratory of Field Crops Department of Agriculture Faculty in Trakya University, Tekirdağ (Turkey. Five different clovers [Persian (Trifolium resupinatum L. var. majus Boiss., Mediterranean (T. spumosum L., narrow-leaved (T. angustifolium L., hedgehog (T. echinatum M. Bieb. and lappa (T. lappaceum L. clovers] were used. Each plot consisted of 8 rows with a length of 5 m. Row spacing of 30 cm and sowing rate of 10 kg ha-1 were used. Sowing times were on 2.25.2001 and on 2.28.2002. Plots were not irrigated and fertilized after sown and harvest. One cut was taken in both years at 4 growing stages such as pre-bud, pre-bloom, 50% bloom and full-bloom. The central 1 m-2 sections was cut at ground level for dry matter. Approximately 500g samples were dried at 55 °C for 24 hours and stored for one day at room temperature then found dry matter. Crude protein (% was determined by Kjeldahl method.

  12. Estimating impact on clover-grass yield caused by traffic intensities

    DEFF Research Database (Denmark)

    Jørgensen, R N; Green, Ole; Kristensen, Kristian

    2009-01-01

    Steer and a 15 m3 Kimadan slurry tanker on two axels, was used to perform the simulated traffic treatment on the parcels. The different traffic intensities are combinations of different tire pressure (1,0 and 2,5 bar), tire load (3000 and 6000 kg), time of year and number of passes (variating from 0 to 8...... components must be determined.   The objective of this paper was to measure yield affects on clover-grass as a consequence of different traffic intensities. The experiments were carried out in the context of a full scale field trial. A 14 hectare full scale grass-clover field trial with 24 different traffic......). The harvesting procedure was preformed with a Haldrup plot harvester modified with RTK-GPS. This paper shows the initial results from measuring the yield affects References M.A. Hamza, M.A.; Anderson, W.K 2005. Soil compaction in cropping systems: A review of the nature, causes and possible solutionsRaper , R...

  13. Morphological traits and yield of red clover (Trifolium pratense L. genotypes with varying inflorescence length

    Directory of Open Access Journals (Sweden)

    Tadeusz Zając

    2013-12-01

    Full Text Available Two red clover breeding strains with elongated flower heads, developed by one of the authors (H.Góral, were studied for forage and seed yield and compared to the standard cultivar 'Nike'. In addition, six morphological shoot traits were measured and their interrelations were computed. The leaf area index (LAI of successive cuts in two harvest years was determined on the basis of shoot density and leaf area of individual shoots. All three genotypes exhibited a high leaf area on shoots and a very high forage productivity. Among the morphological traits only shoot height could be a good selective criterion because it is easily measured, is significantly associated with shoot weight and shoot leaf area and its variation is low. Both strains, particularly the one with longer inflorescences belong to short-lived red clover forms giving satis factory forage and seed yields in the first harvest year. Depending on the strain the number of seeds per elongated inflorescence in the first harvest year was higher by 92 and 42% compared to that of a standard cultivar.

  14. Forage production in mixed grazing systems of elephant grass with arrowleaf clover or forage peanut

    Directory of Open Access Journals (Sweden)

    Daiane Cristine Seibt

    Full Text Available ABSTRACT Most dairy production systems are pasture-based, usually consisting of sole grass species. This system facilitates pasture management, but results in high production costs, mainly because of nitrogen fertilizers. An alternative to making forage systems more sustainable is to introduce legumes into the pasture. Mixed pastures allow better forage distribution over time and reduce fertilization costs. Thus, the objective of this study was to evaluate, throughout the year, three forage systems (FS: FS1 (control - elephant grass (EG, ryegrass (RG, and spontaneous species (SS; FS2 - EG + RG + SS + arrowleaf clover; and FS3 - EG + RG + SS + forage peanut. Elephant grass was planted in rows spaced 4 m apart. Ryegrass was sown between the EG lines, in the winter. Arrowleaf clover was sown according to the respective treatments and forage peanut was preserved. Evaluation was carried out using Holstein cows. The experiment was arranged in a completely randomized design, with three treatments (FS, and three repetitions (paddocks with repeated measurements (grazing cycles. Forage mass achieved 3.46, 3.80, and 3.91 t ha-1 for the treatments FS1, FS2 and FS3, respectively. The forage systems intercropped with legumes produced the best results.

  15. Effect of digestibility of grass-clover silage and concentrate to forage ratio on methane emission from dairy cows

    DEFF Research Database (Denmark)

    Hellwing, Anne Louise Frydendahl; Weisbjerg, Martin Riis

    and D, respectively. TMR forage DM consisted of 2/3 of one of the respective grass-clover silages and 1/3 maize silage, and concentrate (soya meal and wheat) proportion of DM was 20% (low) or 50% (high). Methane emissions from the cows were measured 20-22 hours in one of four chambers working after...

  16. Influence of arbuscular mycorrhizae on biomass production and nitrogen fixation of berseem clover plants subjected to water stress.

    Directory of Open Access Journals (Sweden)

    Sergio Saia

    Full Text Available Several studies, performed mainly in pots, have shown that arbuscular mycorrhizal symbiosis can mitigate the negative effects of water stress on plant growth. No information is available about the effects of arbuscular mycorrhizal symbiosis on berseem clover growth and nitrogen (N fixation under conditions of water shortage. A field experiment was conducted in a hilly area of inner Sicily, Italy, to determine whether symbiosis with AM fungi can mitigate the detrimental effects of drought stress (which in the Mediterranean often occurs during the late period of the growing season on forage yield and symbiotic N2 fixation of berseem clover. Soil was either left under water stress (i.e., rain-fed conditions or the crop was well-watered. Mycorrhization treatments consisted of inoculation of berseem clover seeds with arbuscular mycorrhizal spores or suppression of arbuscular mycorrhizal symbiosis by means of fungicide treatments. Nitrogen biological fixation was assessed using the 15N-isotope dilution technique. Arbuscular mycorrhizal symbiosis was able to mitigate the negative effect of water stress on berseem clover grown in a typical semiarid Mediterranean environment. In fact, under water stress conditions, arbuscular mycorrhizal symbiosis resulted in increases in total biomass, N content, and N fixation, whereas no effect of crop mycorrhization was observed in the well-watered treatment.

  17. A study of the wet deposit and foliar uptake of iodine and strontium on rye-grass and clover

    International Nuclear Information System (INIS)

    Angeletti, Livio; Levi, Emilio; Commission of the European Communities, Ispra

    1977-12-01

    Foliar uptake of iodine and strontium by rye-grass and clover was studied as a function of aspersion intensities. At the same time, the contribution of root sorption to foliar uptake was measured. The effective half-lives of radionuclides of standing and harvested grass were also determined together with their uptake under the action of demineralized water aspersion [fr

  18. Evaluation of physical structure value in spring-harvested grass/clover silage and hay fed to heifers

    DEFF Research Database (Denmark)

    Schulze, A.K.S.; Nørgaard, P.; Byskov, M.V.

    2015-01-01

    The physical structure value of conserved grass/clover forages of spring harvest was evaluated by assessing effects of harvest time, conservation method, iNDF/NDF ratio and NDF intake (NDFI) per kg BW on chewing activity and fecal particle size in dairy heifers. A mixed sward consisting of ryegrass...

  19. Nitrate leaching in maize after cultivation of differently managed grass-clover leys on coarse sand in Denmark

    DEFF Research Database (Denmark)

    Hansen, Elly Møller; Eriksen, Jørgen

    2016-01-01

    When grass-clover leys have been ploughed nitrate leaching may increase. However, management of leys before or after ploughing may affect the leaching risk. We examined the effect of cultivating a six year ley, which the last two years had been treated differently (grazing only; spring cut follow...

  20. Influence of arbuscular mycorrhizae on biomass production and nitrogen fixation of berseem clover plants subjected to water stress.

    Science.gov (United States)

    Saia, Sergio; Amato, Gaetano; Frenda, Alfonso Salvatore; Giambalvo, Dario; Ruisi, Paolo

    2014-01-01

    Several studies, performed mainly in pots, have shown that arbuscular mycorrhizal symbiosis can mitigate the negative effects of water stress on plant growth. No information is available about the effects of arbuscular mycorrhizal symbiosis on berseem clover growth and nitrogen (N) fixation under conditions of water shortage. A field experiment was conducted in a hilly area of inner Sicily, Italy, to determine whether symbiosis with AM fungi can mitigate the detrimental effects of drought stress (which in the Mediterranean often occurs during the late period of the growing season) on forage yield and symbiotic N2 fixation of berseem clover. Soil was either left under water stress (i.e., rain-fed conditions) or the crop was well-watered. Mycorrhization treatments consisted of inoculation of berseem clover seeds with arbuscular mycorrhizal spores or suppression of arbuscular mycorrhizal symbiosis by means of fungicide treatments. Nitrogen biological fixation was assessed using the 15N-isotope dilution technique. Arbuscular mycorrhizal symbiosis was able to mitigate the negative effect of water stress on berseem clover grown in a typical semiarid Mediterranean environment. In fact, under water stress conditions, arbuscular mycorrhizal symbiosis resulted in increases in total biomass, N content, and N fixation, whereas no effect of crop mycorrhization was observed in the well-watered treatment.

  1. Does introduction of clover in an agricultural grassland affect the food base and functional diversity of Collembola?

    NARCIS (Netherlands)

    Annibale, D' Alessandra; Sechi, Valentina; Larsen, Thomas; Christensen, Søren; Krogh, Paul Henning; Eriksen, Jørgen

    2017-01-01

    Introduction of legumes (i.e. white clover) in agricultural grasslands is a common practice to improve yields, but how this affects soil fauna populations, particularly mesofauna, is still poorly understood. We investigated taxonomical and functional differences of Collembola communities between

  2. Uptake and Distribution of Added Selenite and Selenate by Barley and Red Clover as Influenced by Sulphur

    DEFF Research Database (Denmark)

    Nielsen, Gunnar Gissel

    1973-01-01

    The uptake of added Se from selenite and selenate by barley and red clover was investigated in a pot experiment. Much more of selenate than of selenite was taken up but the Se concentrations in the plants declined more with time when selenate was the source. Increasing sulphate addition to the soil...

  3. Environmental life cycle assessments of producing maize, grass-clover, ryegrass and winter wheat straw for biorefinery

    DEFF Research Database (Denmark)

    Parajuli, Ranjan; Kristensen, Ib Sillebak; Knudsen, Marie Trydeman

    2017-01-01

    The aim of this study is to assess the potential environmental impacts of producing maize, grass-clover, ryegrass, and straw from winter wheat as biomass feedstocks for biorefinery. The Life Cycle Assessment (LCA) method included the following impact categories: Global Warming Potential (GWP100),...

  4. A note on the effects of paddock size on the white clover content of swards grazed by sheep

    NARCIS (Netherlands)

    Wolf, de P.L.; Schulte, R.P.O.; Lantinga, E.A.

    2004-01-01

    The maintenance of a high white clover content in mixed swards under sheep grazing has been a challenge to date. This paper presents the results of an experiment in which the effect of the length of a grazing period on the botanical composition of a mixed sward was studied. Paddocks ranging in size

  5. White clover fractions as protein source for monogastrics: dry matter digestibility and protein digestibility-corrected amino acid scores.

    Science.gov (United States)

    Stødkilde, Lene; Damborg, Vinni K; Jørgensen, Henry; Laerke, Helle N; Jensen, Søren K

    2018-05-01

    The present study aimed to evaluate the use of white clover as an alternative protein source for monogastrics. White clover plant and leaves were processed using a screw-press resulting in a solid pulp and a juice from which protein was acid-precipitated. The chemical composition of all fractions was determined and digestibility of dry matter (DM) and protein was assessed in an experiment with growing rats. Protein concentrates were produced with crude protein (CP) contents of 451 g kg -1 and 530 g kg -1 DM for white clover plant and leaves, respectively, and a pulp with CP contents of 313 and 374 g kg -1 DM from plant and leaves, respectively. The amino acid composition ranged from 4.72 to 6.49 g per 16 g of nitrogen (N) for lysine, 1.82-2.6 g per 16 g N for methionine and cysteine, and 3.66-5.24 g per 16 g N for threonine. True faecal digestibility of protein varied from 0.81 to 0.88, whereas DM digestibility was in the range 0.72-0.80. Methionine and cysteine were found to be limiting in all fractions, regardless of the reference group used. A high digestibility of white clover protein was found irrespective of the physical fractionation. Together with a well-balanced amino acid composition, this makes white clover a promising protein source for monogastrics. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  6. Soil properties and clover establishment six years after surface application of calcium-rich by-products

    Energy Technology Data Exchange (ETDEWEB)

    Ritchey, K.D.; Belesky, D.P.; Halvorson, J.J. [USDA ARS, Beaver, WV (US). Appalachian Farming Systems Research Center

    2004-12-01

    Calcium-rich soil amendments can improve plant growth by supplying Ca and reducing detrimental effects of soil acidity, but solubility and neutralizing capacity of Ca sources vary. Our objectives were to evaluate effects of calcitic dolomite and several coal combustion by-products on soil properties at various depths 6 yr after surface application and their influence on grass-clover herbage accumulation. Calcium and Mg soil amendments were surface-applied to an acidic grassland in 1993, and orchardgrass (Dactylis glomerata L.) and tall fescue (Lolium arundinaceum (Schreb.) Darbyshire) were oversown in 1994. In 1998, amendment treatment plots were split to accommodate sod seeding with red clover (Trifolium pratense L.) or white clover (T. repens L.) as well as a nonseeded control. No N fertilizer was applied after sod seeding. Six years after amendment application, reductions in soil Al and Mn and increases in Ca and pH from 4654 kg ha{sup -1} calcitic dolomite, 15 000 kg ha{sup -1} fluidized bed combustion residue, or 526 kg ha{sup -1} MgO amendment were greatest in the surface 2.5 cm while rates of gypsum as high as 32 000 kg ha{sup -1} left little residual effect except for decreases in Mg. Percentage clover in the sward tripled as pH increased from 4.3 to 5.0 while herbage mass increased 75% as clover percentage increased. Herbage mass was generally more closely correlated with properties of soil samples collected from the surface 2.5 cm than from deeper samples.

  7. Utilization of kura clover-reed canarygrass silage versus alfalfa silage by lactating dairy cows.

    Science.gov (United States)

    Kammes, K L; Heemink, G B H; Albrecht, K A; Combs, D K

    2008-08-01

    The mixture of kura clover (Trifolium ambiguum M. Bieb.) and reed canarygrass (Phalaris arundinacea L.) has proven to be extremely persistent in the northern United States, but information about dairy cow performance on this mixture is lacking. Twenty lactating Holstein cows were used in a crossover design to compare dry matter (DM) intake and milk production from diets containing kura clover-reed canarygrass silage (KRS) or alfalfa (Medicago sativa L.) silage (AS). Forages were cut, wilted, ensiled in horizontal plastic bags, and allowed to ferment for at least 50 d before beginning the feeding experiment. The KRS was approximately 40% kura clover and 60% reed canarygrass. Treatments were total mixed rations formulated with either 57% of total DM from 1) AS or 2) KRS. Experimental periods were 28 d, with the first 14 d for diet adaptation and the last 14 d for measurement of intake and milk production. The neutral detergent fiber (NDF) concentrations of AS and KRS were 37.3 and 47.3%, respectively. The fermentation analyses indicated that both silages underwent a restricted fermentation, producing primarily lactic acid and some acetic acid. Dry matter intake (24.2 vs. 22.8 kg) and 4% fat-corrected milk (32.8 vs. 30.9 kg) were significantly higher for cows fed AS than for cows fed KRS. Cows consumed less NDF (6.7 vs. 8.0 kg) and less digestible NDF (3.0 vs. 4.4 kg) when fed AS diets compared with KRS diets, but the pool of ruminally undegraded NDF was similar (3.7 kg) between diets. Cows produced 1.5 kg of milk/kg of DM consumed regardless of the diet, indicating that digestible NDF of KRS was utilized with similar efficiency as the cell wall constituents of AS, but the intake of cows fed KRS may have been limited by rumen fill. Milk fat concentration tended to be higher for cows fed AS, but the milk true protein concentration and yields of fat and protein did not differ by treatment. Milk urea nitrogen content was higher when cows consumed AS (16.4 mg/ d

  8. Red Clover

    Science.gov (United States)

    ... cancer or other hormone-sensitive cancers. Keep in Mind Tell all your health care providers about any ... Privacy and Policies Accessibility en Español FOIA Site Map Contact Us U.S. Department of Health & Human Services , ...

  9. Meat goat kids finished on alfalfa, red clover or orchardgrass pastures: carcass merit and meat quality.

    Science.gov (United States)

    Turner, K E; Cassida, K A; Zerby, H N

    2014-12-01

    This experiment was conducted in 2005-2007 to evaluate carcass and chevon (goat meat) quality parameters when meat-goat kids (n=72) were finished on alfalfa (Medicago sativa L; ALF); red clover (Trifolium pratense L.; RCG); or orchardgrass (Dactylis glomerata L.; OGR) pastures. Carcass conformation score was greater (P=0.08) when meat-goat kids were finished on ALF compared to OGR with RCG intermediate. Chevon meat samples from goats finished on the three pasture treatments did not differ in ash, intramuscular fat, or crude protein content or in concentrations of omega6 and omega3 fatty acids, or the omega6 to omega3 ratio. Goats finished on OGR had higher (Pmeat-goat kids finished on ALF, RCG, or ORG produced desirable carcass weights for most niche markets in the USA. Chevon is a low-fat meat option with high desirable fatty acids for human diets. Published by Elsevier Ltd.

  10. Measured and simulated performance of Compton-suppressed TIGRESS HPGe clover detectors

    Science.gov (United States)

    Schumaker, M. A.; Hackman, G.; Pearson, C. J.; Svensson, C. E.; Andreoiu, C.; Andreyev, A.; Austin, R. A. E.; Ball, G. C.; Bandyopadhyay, D.; Boston, A. J.; Chakrawarthy, R. S.; Churchman, R.; Drake, T. E.; Finlay, P.; Garrett, P. E.; Grinyer, G. F.; Hyland, B.; Jones, B.; Maharaj, R.; Morton, A. C.; Phillips, A. A.; Sarazin, F.; Scraggs, H. C.; Smith, M. B.; Valiente-Dobón, J. J.; Waddington, J. C.; Watters, L. M.

    2007-01-01

    Tests of the performance of a 32-fold segmented HPGe clover detector coupled to a 20-fold segmented Compton-suppression shield, which form a prototype element of the TRIUMF-ISAC Gamma-Ray Escape-Suppressed Spectrometer (TIGRESS), have been made. Peak-to-total ratios and relative efficiencies have been measured for a variety of γ-ray energies. These measurements were used to validate a GEANT4 simulation of the TIGRESS detectors, which was then used to create a simulation of the full 12-detector array. Predictions of the expected performance of TIGRESS are presented. These predictions indicate that TIGRESS will be capable, for single 1 MeV γ rays, of absolute detection efficiencies of 17% and 9.4%, and peak-to-total ratios of 54% and 61% for the "high-efficiency" and "optimized peak-to-total" configurations of the array, respectively.

  11. Response of irradiated Egyptian clover (Trifolium alexandrinum L.) to water stress

    International Nuclear Information System (INIS)

    Hamideldin, N; Hussein, O.S

    2009-01-01

    Egyptian clover (Trifolium alexandrinum L.) is one of the main leguminous forage crops in Egypt. Gamma rays represent one of the important physical activators for growth and productivity. The seed were irradiated using three doses of gamma radiation (100,200 and 300 Gy) and two levels of irrigation (10 and 20 days) after germination. Gamma radiation treatments stimulated growth criteria in normal irrigation treatment, while drought irrigation treatment stunted these criteria. The most significant dose was 100 Gy, which stimulated growth criteria, chlorophylls a , b, total chlorophyll and carotenoids in both irrigation treatments used and growth hormones under drought condition. Drought stress and gamma irradiation increase amino acid content and caused change in band concentration of esterase and peroxidase isozyme .Also, 13 protein bands appeared only in treated plant with 100 Gy under drought condition. It could be concluded that the low dose (100 Gy) of gamma irradiation release the effect of drought stress.

  12. New uses of clover-grass mixtures in the structure of fodder crops on arable land

    Directory of Open Access Journals (Sweden)

    Jiří Sláma

    2010-01-01

    Full Text Available The use of clover-grasses in the structure of fodder crops grown on arable soil, especially those with intergeneric hybrids as the main component part, could avert the negative current trend, i.e. further decreasing the area of perennial fodder plants or fodder crops as a whole on arable soil. They have an irreplaceable role in crop sequences and in preserving the cultural character of the countryside, above all due to the fact that they improve soil fertility and microbial life in the soil and that they have an excellent pre-produce value, and, at the same time, they are applied in various farming systems (both conventional and ecological and in various climatic conditions, and agricultural businesses are well equipped for growing, harvesting and storing them. In the Czech Republic, the area of fodder crops grown on arable soil was decreased from 1,019.9 thousand hectares to mere 396.7 thousand hectares between 1980 and 2009, which is 15.6 % of the total area of arable soil whereas perennial fodder plants only take up 8.5 %. Fodder from clover crops and clover-grass growths on arable soil are one of the main resources of voluminous fodder for dairy cows. Most of this fodder is preserved through a fermentation process (silages, hay storage; a smaller part is fed as fresh fodder, or serves for production of hay. Silages made with perennial fodder plants are the most important source of both proteins and other nutrients for ruminants, especially for high-yielding milch cows. The basis of fodder production systems are the conservative elements of the landscape area (geomorphology in combination with the progressive elements (weather conditions, plants and human labour and relict ones, the representative of which is the soil. The fodder production systems in Europe are divided into five main fodder production zones. From this point of view, the areas where short-term clover-grass mixtures are grown on arable soil could be classed with Zone 4, i

  13. Sexual polyploidization in red clover Poliploidização sexual em trevo vermelho

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    Carine Simioni

    2006-02-01

    Full Text Available Because sexual polyploidization broadens genetic basis and supply plant breeders with more variability for the selection process, it can be useful in red clover breeding. This paper reports results of three crossing cycles, starting from a parental generation of tetraploid red clover plants (female parent, and diploids from the Quiñiqueli cultivar, selected for production of more than 1% of giant pollen grains (male parent aiming to obtain tetraploid plants to be used in red clover breeding programs. Crosses in the next generations were performed by mutual cross-pollinations. Chromosome number chimerism and high pollen sterility were detected in F1, F2 and F3, but there was a trend towards increasing seed production and seed viability along the generations, probably due to successful competition between fertile and sterile gametes. The identification of fertile triploids, as well as their recurrent formation along the generations, indicates that triploid block is not complete in red clover, and that triploids may be successfully used as a bridge for the production of sexual polyploids.Porque a poliploidização sexual amplia a base genética e proporciona aos melhoristas maior variabilidade para o processo de seleção, ela pode ser uma ferramenta útil ao melhoramento de trevo vermelho. Com o objetivo de obter plantas tetraplóides que possam ser utilizadas em programas de melhoramento de trevo vermelho, este trabalho relata resultados de três ciclos de cruzamentos, partindo de uma população parental de plantas tetraplóides de trevo vermelho, como genitores femininos, e de diplóides da cultivar Quiñiqueli, selecionados para produção de mais de 1% de grãos de pólen gigantes, como genitores masculinos. Nas outras gerações, os cruzamentos foram realizados por polinizações cruzadas mútuas. Quimerismo para número cromossômico e alta esterilidade de pólen foram detectados em F1 , F2 e F3, mas houve uma tendência para aumento da

  14. Interaction of pleuromutilin derivatives with the ribosomal peptidyl transferase center

    DEFF Research Database (Denmark)

    Long, K. S.; Hansen, L. K.; Jakobsen, L.

    2006-01-01

    Tiamulin is a pleuromutilin antibiotic that is used in veterinary medicine. The recently published crystal structure of a tiamulin-50S ribosomal subunit complex provides detailed information about how this drug targets the peptidyl transferase center of the ribosome. To promote rational design...... mutant strain is resistant to tiamulin and pleuromutilin, but not valnemulin, implying that valnemulin is better able to withstand an altered rRNA binding surface around the mutilin core. This is likely due to additional interactions made between the valnemulin side chain extension and the rRNA binding...

  15. Ozone and Water Stress: Effects on the Behaviour of Two White Clover Biotypes

    Directory of Open Access Journals (Sweden)

    Massimo Fagnano

    Full Text Available ozone pollution, water stress, stomata conductance, ozone uptake, clover, OTC.Ozone is a strong oxidizing pollutant which derives by alteration of the photolytic NOx cycle and it accumulates in the troposphere spreading in rural areas and therefore determining injuries on natural vegetation and crops. Since its penetration occurs mainly through stomata, all factors which alter plant-atmosphere relations could be able to modify plant response to ozone. Interaction between ozone and water stress in Mediterranean environment was studied on ozone resistant and sensitive biotypes of white clover, which were grown in charcoal filtered and notfiltered Open Top Chambers in factorial combination with different levels of water supply. Measurements of biomass, leaf area and stomatal conductance were made during the growth period. Ozone injuries were estimated as not-filtered/filtered OTC yield ratio; the stomatal flux of ozone was estimated multiplying stomata conductance x diffusivity ratio between ozone and water vapour (0.613 x ozone hourly concentrations. The hourly values of ozone uptake were cumulated throughout the cropping periods of the two years. In the sensitive biotype, water stress reduced yield losses due to ozone from 38% to 22%, as well as yield losses due to water stress were reduced by the presence of ozone from 43% to 29%, while no interaction between ozone and water stress was observed in the resistant biotype. Biomass yield losses of the sensitive biotype were strictly correlated to cumulated ozone uptake (R2 = 0.99, while biomass yield losses of the resistant biotype were not affected by the ozone fluxes variations created by the treatments. Flux based models could better estimate yield losses due to ozone in Mediterranean environments in which other stresses could be contemporary present; therefore, the new European directives might replace the actual thresholds based

  16. Quantitative historical change in bumblebee (Bombus spp. assemblages of red clover fields.

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    Yoko L Dupont

    Full Text Available BACKGROUND: Flower visiting insects provide a vitally important pollination service for many crops and wild plants. Recent decline of pollinating insects due to anthropogenic modification of habitats and climate, in particular from 1950's onwards, is a major and widespread concern. However, few studies document the extent of declines in species diversity, and no studies have previously quantified local abundance declines. We here make a quantitative assessment of recent historical changes in bumblebee assemblages by comparing contemporary and historical survey data. METHODOLOGY/PRINCIPAL FINDINGS: We take advantage of detailed, quantitative historical survey data from the 1930's on bumblebee (Bombus spp. abundances and species composition in red clover (Trifolium pratense fields, an important floral resource and an attractant of all bumblebee species. We used the historical survey data as a pre-industrialization baseline, and repeated the same sampling protocol at nearly the same localities at present, hence setting up a historical experiment. We detected historical changes in abundances (bees/m(2 of both workers (the "pollinatory units" and queens (effective population size, in addition to species composition. In particular, long-tongued bumblebee species showed consistent and dramatic declines in species richness and abundances throughout the flowering season of red clover, while short-tongued species were largely unaffected. Of 12 Bombus species observed in the 1930's, five species were not observed at present. The latter were all long-tongued, late-emerging species. CONCLUSIONS/SIGNIFICANCE: Because bumblebees are important pollinators, historical changes in local bumblebee assemblages are expected to severely affect plant reproduction, in particular long-tubed species, which are pollinated by long-tongued bumblebees.

  17. Forage mass and the nutritive value of pastures mixed with forage peanut and red clover

    Directory of Open Access Journals (Sweden)

    Ricardo Lima de Azevedo Junior

    2012-04-01

    Full Text Available The objective of this research was to estimate three pasture-based systems mixed with elephantgrass + spontaneous growth species, annual ryegrass, for pasture-based system 1; elephantgrass + spontaneous growth species + forage peanut, for pasture-based system 2; and elephantgrass + spontaneous growth species + annual ryegrass + red clover, for pasture-based system 3. Elephantgrass was planted in rows 4 m apart from each other. During the cool-season, annual ryegrass was sown in the alleys between the rows of elephantgrass; forage peanut and red clover were sown in the alleys between the elephantgrass according to the respective treatment. The experimental design was totally randomized in the three treatments (pasture-based systems, two replicates (paddocks in completely split-plot time (grazing cycles. Holstein cows receiving 5.5 kg-daily complementary concentrate feed were used in the evaluation. Pre-grazing forage mass, botanical composition and stocking rate were evaluated. Samples of simulated grazing were collected to analyze organic matter (OM, neutral detergent fiber (NDF, crude protein (CP and organic matter in situ digestibility (OMISD. Nine grazing cycles were performed during the experimental period (341 days. The average dry matter values for pre-grazing and stocking rate were 3.34; 3.46; 3.79 t/ha, and 3.28; 3.34; 3.60 AU/ha for each respective pasture-based system. Similar results were observed between the pasture-based systems for OM, NDF, CP and OMISD. Considering forage mass, stocking rate and nutritive value, the pasture-based system intercropped with forage legumes presented better performance.

  18. Medium-term response of microbial community to rhizodeposits of white clover and ryegrass and tracing of active processes induced by 13C and 15N labelled exudates

    DEFF Research Database (Denmark)

    Kusliene, Gedrime; Rasmussen, Jim; Kuzyakov, Yakov

    2014-01-01

    and actinomycetes was unaffected by plant species, but pool of Gram-negative and Gram-positive bacteria was greater under white clover at the 10 percent significance level. In the short term, microorganisms more actively utilised fresh exudates (13C-labelled) of ryegrass than of white clover. We expected ryegrass...... microbial groups in soil under white clover (Trifolium repens L.) and ryegrass (Lolium perenne L.) following leaf-labelling with 13C-bicarbonate and 15N-urea. In this way microbial N and 15N and the composition of PLFAs reflect the medium-term (two months) response of microorganisms to rhizodeposits......, whereas the 13C-label of the PLFAs reflects the short-term (one week) utilisation of root exudates following labelling of shoots. In the medium term, microbial biomass N and 15N were greater under the ryegrass, whereas total PLFA was higher under white clover. The relative abundance of fungi...

  19. A glutathione s-transferase confers herbicide tolerance in rice

    Directory of Open Access Journals (Sweden)

    Tingzhang Hu

    2014-07-01

    Full Text Available Plant glutathione S-transferases (GSTs have been a focus of attention due to their role in herbicide detoxification. OsGSTL2 is a glutathione S-transferase, lambda class gene from rice (Oryza sativa L.. Transgenic rice plants over-expressing OsGSTL2 were generated from rice calli by the use of an Agrobacterium transformation system, and were screened by a combination of hygromycin resistance, PCR and Southern blot analysis. In the vegetative tissues of transgenic rice plants, the over-expression of OsGSTL2 not only increased levels of OsGSTL2 transcripts, but also GST and GPX expression, while reduced superoxide. Transgenic rice plants also showed higher tolerance to glyphosate and chlorsulfuron, which often contaminate agricultural fields. The findings demonstrate the detoxification role of OsGSTL2 in the growth and development of rice plants. It should be possible to apply the present results to crops for developing herbicide tolerance and for limiting herbicide contamination in the food chain.

  20. Interaction of Pleuromutilin Derivatives with the Ribosomal Peptidyl Transferase Center

    Science.gov (United States)

    Long, Katherine S.; Hansen, Lykke H.; Jakobsen, Lene; Vester, Birte

    2006-01-01

    Tiamulin is a pleuromutilin antibiotic that is used in veterinary medicine. The recently published crystal structure of a tiamulin-50S ribosomal subunit complex provides detailed information about how this drug targets the peptidyl transferase center of the ribosome. To promote rational design of pleuromutilin-based drugs, the binding of the antibiotic pleuromutilin and three semisynthetic derivatives with different side chain extensions has been investigated using chemical footprinting. The nucleotides A2058, A2059, G2505, and U2506 are affected in all of the footprints, suggesting that the drugs are similarly anchored in the binding pocket by the common tricyclic mutilin core. However, varying effects are observed at U2584 and U2585, indicating that the side chain extensions adopt distinct conformations within the cavity and thereby affect the rRNA conformation differently. An Escherichia coli L3 mutant strain is resistant to tiamulin and pleuromutilin, but not valnemulin, implying that valnemulin is better able to withstand an altered rRNA binding surface around the mutilin core. This is likely due to additional interactions made between the valnemulin side chain extension and the rRNA binding site. The data suggest that pleuromutilin drugs with enhanced antimicrobial activity may be obtained by maximizing the number of interactions between the side chain moiety and the peptidyl transferase cavity. PMID:16569865

  1. A novel method for screening the glutathione transferase inhibitors

    Directory of Open Access Journals (Sweden)

    Węgrzyn Grzegorz

    2009-03-01

    Full Text Available Abstract Background Glutathione transferases (GSTs belong to the family of Phase II detoxification enzymes. GSTs catalyze the conjugation of glutathione to different endogenous and exogenous electrophilic compounds. Over-expression of GSTs was demonstrated in a number of different human cancer cells. It has been found that the resistance to many anticancer chemotherapeutics is directly correlated with the over-expression of GSTs. Therefore, it appears to be important to find new GST inhibitors to prevent the resistance of cells to anticancer drugs. In order to search for glutathione transferase (GST inhibitors, a novel method was designed. Results Our results showed that two fragments of GST, named F1 peptide (GYWKIKGLV and F2 peptide (KWRNKKFELGLEFPNL, can significantly inhibit the GST activity. When these two fragments were compared with several known potent GST inhibitors, the order of inhibition efficiency (measured in reactions with 2,4-dinitrochlorobenzene (CDNB and glutathione as substrates was determined as follows: tannic acid > cibacron blue > F2 peptide > hematin > F1 peptide > ethacrynic acid. Moreover, the F1 peptide appeared to be a noncompetitive inhibitor of the GST-catalyzed reaction, while the F2 peptide was determined as a competitive inhibitor of this reaction. Conclusion It appears that the F2 peptide can be used as a new potent specific GST inhibitor. It is proposed that the novel method, described in this report, might be useful for screening the inhibitors of not only GST but also other enzymes.

  2. Acrolein-detoxifying isozymes of glutathione transferase in plants.

    Science.gov (United States)

    Mano, Jun'ichi; Ishibashi, Asami; Muneuchi, Hitoshi; Morita, Chihiro; Sakai, Hiroki; Biswas, Md Sanaullah; Koeduka, Takao; Kitajima, Sakihito

    2017-02-01

    Acrolein is a lipid-derived highly reactive aldehyde, mediating oxidative signal and damage in plants. We found acrolein-scavenging glutathione transferase activity in plants and purified a low K M isozyme from spinach. Various environmental stressors on plants cause the generation of acrolein, a highly toxic aldehyde produced from lipid peroxides, via the promotion of the formation of reactive oxygen species, which oxidize membrane lipids. In mammals, acrolein is scavenged by glutathione transferase (GST; EC 2.5.1.18) isozymes of Alpha, Pi, and Mu classes, but plants lack these GST classes. We detected the acrolein-scavenging GST activity in four species of plants, and purified an isozyme showing this activity from spinach (Spinacia oleracea L.) leaves. The isozyme (GST-Acr), obtained after an affinity chromatography and two ion exchange chromatography steps, showed the K M value for acrolein 93 μM, the smallest value known for acrolein-detoxifying enzymes in plants. Peptide sequence homology search revealed that GST-Acr belongs to the GST Tau, a plant-specific class. The Arabidopsis thaliana GST Tau19, which has the closest sequence similar to spinach GST-Acr, also showed a high catalytic efficiency for acrolein. These results suggest that GST plays as a scavenger for acrolein in plants.

  3. Characterisation of a compton suppressed clover detector for high energy gamma rays (5 MeV ≤ E ≤ 11 MeV)

    International Nuclear Information System (INIS)

    Saha Sarkar, M.; Kshetri, Ritesh; Raut, Rajarshi; Mukherjee, A.; Goswami, A.; Ray, S.; Basu, P.; Majumder, H.; Bhattacharya, S.; Dasmahapatra, B.; Sinha, Mandira; Ray, Maitreyee

    2004-01-01

    The Clover detectors in their add back mode have been seen to be excellent tools for detecting high energy gamma rays (≥ 2 MeV). Recently studies were carried out on the characteristics of a Compton suppressed Clover germanium detector up to 5 MeV using a radioactive 66 Ga (T 1/2 =9.41 h) source for the first time

  4. Environmental Impact Research Program: White Clover (Trifolium repens). Section 7.3.5, US Army Corps of Engineers Wildlife Resources Management Manual.

    Science.gov (United States)

    1986-07-01

    types where they are common (Kohn and Mooty 1971). Rumen analyses of white-tailed deer in Montana showed that forbs comprised 80% of summer samples...frequency of occurrence in deer rumens collected from January through March in the Northern Black Hills. Bluegrass- white clover meadows are also...CAUTIONS AND LIMITATIONS White clover, like most legumes, occasionally causes bloat in animals, particularly cattle and sheep that have overfilled

  5. Effects of feeding level and NDF content of grass-clover silages on chewing activity, fecal particle size and NDF digestibility in dairy heifers

    DEFF Research Database (Denmark)

    Schulze, Anne-Katrine Skovsted; Weisbjerg, Martin Riis; Nørgaard, Peder

    2014-01-01

    intake (PRumination time per kg DM intake (Pcontents (Prumination with greater...... NDF content (Prumination time increased with greater NDF content (Pcontent (P...The objective of this study was to assess effects of feed intake and NDF content of highly digestible grass-clover silage on chewing behavior, fecal particle size distribution and apparent digestibility in restrictively fed heifers. Four grass-clover silages (Lolium perenne, Trifolium pratense...

  6. Metabolisable protein supply to lactating dairy cows increased with increasing dry matter concentration in grass-clover silage

    DEFF Research Database (Denmark)

    Johansen, Marianne; Hellwing, Anne Louise Frydendahl; Lund, Peter

    2017-01-01

    The aim of this experiment was to study the effect of increased dry matter (DM) concentration in grass-clover silage, obtained by extending the pre-wilting period before ensiling, on the amount of metabolisable protein (MP) supplied to lactating dairy cows. Spring growth and first regrowth of grass...... and faeces, respectively, were collected over 94 h to cover the diurnal variation, pooled, and subsequently analysed. Rumen fluid was collected in same sampling procedure. To estimate the duodenal flow of microbial protein, microbes were isolated from the rumen and analysed for amino acids (AA) and purines...... flow of AA. The higher duodenal flow of AA derived from a lower rumen degradation of feed protein and a tendency towards a higher microbial synthesis in the rumen. Fibre digestibility and CH4 production were not affected by silage DM concentration. In conclusion, MP concentration in grass-clover silage...

  7. Effects of soil properties and P fertilizers on trace element uptake of red clover in a pot experiment

    International Nuclear Information System (INIS)

    Osztoics, E.; Bujtas, K.

    1999-01-01

    The impacts of superphosphate and Algerian phosphate rock and their various application rates on soil pH and on the availability of trace elements by red clover were studied in a pot experiment on several types of acidic soils from the Carpathian basin. The differences among the soils' original pH and texture, and those differences in soil pH, which resulted from the application of different P forms and rotes were reflected in the Mn, Ni, Al, Co, Sr, Cd and Cr contents of red clover. Plant concentrations of those elements were smaller on the slightly acidic than on the strongly and extremely strongly acidic soils. Elemental concentrations were generally higher when there was less time between two cuts, and decreased in the later cuts. Refs. 11 (author)

  8. Assessment of Protective Effect of Some Modern Agrochemicals against Ozone-Induced Stress in Sensitive Clover and Tobacco Cultivars.

    Science.gov (United States)

    Blum, Oleg; Didyk, Nataliya; Pavluchenko, Nataliya; Godzik, Barbara

    2011-01-01

    Some modern agrochemicals with antioxidant potential were tested for their protective effect against ozone injury using clover and tobacco ozone-sensitive cultivars as model plants subjected to ambient ozone at two sites (Kyiv city in Ukraine and Szarów village in Poland). All used agrochemicals showed partial protective effects against ozone injury on clover and tobacco. Conducted studies confirmed the effectiveness of modern fungicides belonging to strobilurin group as protectants of sensitive crops against ozone damage. The effectiveness of new growth regulators "Emistym C" and "Agrostymulin" was showed for the first time. Out of the studied agrochemicals, fungicide "Strobi" and natural growth regulator "Emistym C" demonstrated the best protective effects. These agrochemicals present promise for further studies of their possible utilization for enhancement of ozone tolerance of sensitive crops.

  9. Optimization of a protocol for direct organogenesis of red clover (Trifolium pratense L. meristems for breeding purposes

    Directory of Open Access Journals (Sweden)

    JUAN C CARRILLO

    2004-01-01

    Full Text Available A series of experiments were carried out in order to optimize a protocol for the direct organogenesis of Chilean red clover germplasm. A range of cultivars were used to analyze the effect of explant source (crown or stem meristems of vegetative plants, culture media and plant growth regulators. Our findings showed that stem meristems were easier to obtain, presented lower levels of contamination and a better development than crown meristems. The L2 medium showed better results than B5 and MS media for the cultivars and experimental lines studied. L2 medium supplemented with 0.003 mg/l of 4-amino-3,5,6-trichloropicolinic acid and 1.0 mg/l of 6-benzylaminopurine gave consistently better results and will be applied in our breeding program to propagate, maintain and eliminate viruses from elite red clover clones.

  10. The beneficial effect of dual inoculation of vesicular-arbuscular mycorrhizae + rhizobium on growth of white clover

    Directory of Open Access Journals (Sweden)

    Lin, XG.

    1993-01-01

    Full Text Available Investigation on the effect of phosphorus on vesicular-arbuscular mycorrhizal infection, and dual inoculation of vesicular-arbuscular mycorrhizae + rhizobium on growth of white clover under field microplots and pot experiments was conducted on fluvo-aquic soils of semi-arid region in north China. The results showed that 60 kg P205 ha in form of superphosphate was the most favorable phosphorus level for vesicular-arbuscular mycorrhizal infection ; mycorrhizal infection, nodulation, dry weight of shoots and roots, total uptake of nitrogen, phosphorus and other elements, the final yields and recovery of phosphorus of white clover were significantly increased by vesicular-arbuscular mycorrhizal inoculation and dual inoculation with vesicular-arbuscular mycorrhizal fungi and rhizobium. The highest response of inoculation was obtained by adding fertilizer phosphorus at the level of 60 kg P205 ha in form of superphosphate.

  11. Effect of Pseudomonas fluorescens and pyoverdine on the phytoextraction of cesium by red clover in soil pots and hydroponics.

    Science.gov (United States)

    Hazotte, Alice; Péron, Olivier; Gaudin, Pierre; Abdelouas, Abdesselam; Lebeau, Thierry

    2018-05-12

    With the aim of improving the phytoextraction rate of cesium (Cs), the effect of Pseudomonas fluorescens ATCC 17400 and its siderophore pyoverdine (PVD) on the uptake of Cs by red clover was studied in soil pots. This work also provides a mechanistic understanding of the Cs-bacteria (or PVD)-illite-plant interactions by using a simplified experimental design, i.e., hydroponics with either Cs in solution or Cs-spiked illite in suspension. For soil spiked with 11.2 mmol kg -1 (1480 mg kg -1 ) of Cs, 0.43% of total Cs was taken up by red clover in 12 days (119 μmol g -1 (16 mg g -1 ) of Cs dry matter in roots and 40 μmol g -1 (5 mg g -1 ) in shoots). In hydroponics with Cs in solution (0.1 mmol L -1 or 13 mg L -1 ), 75% of Cs was taken up vs. only 0.86% with Cs-spiked illite suspension. P. fluorescens and PVD did not increase Cs concentrations in aboveground parts and roots of red clover and even decreased them. The damaging effect of PVD on red clover growth was demonstrated with the biomass yielding 66% of the control in soil pots (and 100% mortality after 12 days of exposition) and only 56% in hydroponics (78% with illite in suspension). Nonetheless, PVD and, to a lesser extent, P. fluorescens increased the translocation factor up to a factor of 2.8. This study clearly showed a direct damaging effect of PVD and to a lower extent the retention of Cs by biofilm covering both the roots and illite, both resulting in the lower phytoextraction efficiency.

  12. The beneficial effect of dual inoculation of vesicular-arbuscular mycorrhizae + rhizobium on growth of white clover

    OpenAIRE

    Lin, XG.; Hao, WY.; Wu, TH.

    1993-01-01

    Investigation on the effect of phosphorus on vesicular-arbuscular mycorrhizal infection, and dual inoculation of vesicular-arbuscular mycorrhizae + rhizobium on growth of white clover under field microplots and pot experiments was conducted on fluvo-aquic soils of semi-arid region in north China. The results showed that 60 kg P205 ha in form of superphosphate was the most favorable phosphorus level for vesicular-arbuscular mycorrhizal infection ; mycorrhizal infection, nodulation, dry weight ...

  13. Towards an optimal sampling strategy for assessing genetic variation within and among white clover (Trifolium repens L. cultivars using AFLP

    Directory of Open Access Journals (Sweden)

    Khosro Mehdi Khanlou

    2011-01-01

    Full Text Available Cost reduction in plant breeding and conservation programs depends largely on correctly defining the minimal sample size required for the trustworthy assessment of intra- and inter-cultivar genetic variation. White clover, an important pasture legume, was chosen for studying this aspect. In clonal plants, such as the aforementioned, an appropriate sampling scheme eliminates the redundant analysis of identical genotypes. The aim was to define an optimal sampling strategy, i.e., the minimum sample size and appropriate sampling scheme for white clover cultivars, by using AFLP data (283 loci from three popular types. A grid-based sampling scheme, with an interplant distance of at least 40 cm, was sufficient to avoid any excess in replicates. Simulations revealed that the number of samples substantially influenced genetic diversity parameters. When using less than 15 per cultivar, the expected heterozygosity (He and Shannon diversity index (I were greatly underestimated, whereas with 20, more than 95% of total intra-cultivar genetic variation was covered. Based on AMOVA, a 20-cultivar sample was apparently sufficient to accurately quantify individual genetic structuring. The recommended sampling strategy facilitates the efficient characterization of diversity in white clover, for both conservation and exploitation.

  14. Analysis of glutathione S-transferase (M1, T1 and P1) gene ...

    African Journals Online (AJOL)

    Glutathione S-transferase enzymes are active in detoxifying a wide number of endogenous and exogenous chemical carcinogens and subsequently, are crucial in protecting the DNA. Several studies show some differences in association of glutathione S-transferase M1, T1 and P1 genetic polymorphisms with the risk of ...

  15. From glutathione transferase to pore in a CLIC

    CERN Document Server

    Cromer, B A; Morton, C J; Parker, M W; 10.1007/s00249-002-0219-1

    2002-01-01

    Many plasma membrane chloride channels have been cloned and characterized in great detail. In contrast, very little is known about intracellular chloride channels. Members of a novel class of such channels, called the CLICs (chloride intracellular channels), have been identified over the last few years. A striking feature of the CLIC family of ion channels is that they can exist in a water- soluble state as well as a membrane-bound state. A major step forward in understanding the functioning of these channels has been the recent crystal structure determination of one family member, CLIC1. The structure confirms that CLICs are members of the glutathione S- transferase superfamily and provides clues as to how CLICs can insert into membranes to form chloride channels. (69 refs).

  16. Kunitz Proteinase Inhibitors Limit Water Stress Responses in White Clover (Trifolium repens L.) Plants.

    Science.gov (United States)

    Islam, Afsana; Leung, Susanna; Nikmatullah, Aluh; Dijkwel, Paul P; McManus, Michael T

    2017-01-01

    The response of plants to water deficiency or drought is a complex process, the perception of which is triggered at the molecular level before any visible morphological responses are detected. It was found that different groups of plant proteinase inhibitors (PIs) are induced and play an active role during abiotic stress conditions such as drought. Our previous work with the white clover ( Trifolium repens L.) Kunitz Proteinase Inhibitor ( Tr-KPI ) gene family showed that Tr-KPIs are differentially regulated to ontogenetic and biotic stress associated cues and that, at least some members of this gene family may be required to maintain cellular homeostasis. Altered cellular homeostasis may also affect abiotic stress responses and therefore, we aimed to understand if distinct Tr-PKI members function during drought stress. First, the expression level of three Tr-KPI genes, Tr-KPI1 , Tr-KPI2 , and Tr-KPI5 , was measured in two cultivars and one white clover ecotype with differing capacity to tolerate drought. The expression of Tr-KPI1 and Tr-KPI5 increased in response to water deficiency and this was exaggerated when the plants were treated with a previous period of water deficiency. In contrast, proline accumulation and increased expression of Tr-NCED1 , a gene encoding a protein involved in ABA biosynthesis, was delayed in plants that experienced a previous drought period. RNAi knock-down of Tr-KPI1 and Tr-KPI5 resulted in increased proline accumulation in leaf tissue of plants grown under both well-watered and water-deficit conditions. In addition, increased expression of genes involved in ethylene biosynthesis was found. The data suggests that Tr-KPIs , particularly Tr-KPI5 , have an explicit function during water limitation. The results also imply that the Tr-KPI family has different in planta proteinase targets and that the functions of this protein family are not solely restricted to one of storage proteins or in response to biotic stress.

  17. Reaction of cocksfoot (Dactylis glomerata L. in grass-clover mixture on N fertilization and grazing

    Directory of Open Access Journals (Sweden)

    Marcela Andreata-Koren

    2009-09-01

    Full Text Available Grazing is the most common way of using a hill and mountain areas because of their specific agro-ecological conditions. Cocksfoot is a grass with high productivity and quality, and it is very good for sowing in the sward for grazing. Because of its good adaptability to different growing conditions, especially in very dry and cold areas, it is excellent in relation to some other good grasses, which can not be raised in such areas. The aim of the experiment was to determine effect of N application (0-N0 and 150 kg ha-1 year-1-N150 and rotational grazing by cattle (C and sheep (S, and their interaction on the cocksfoot sown in a mixture of smooth-stalked meadow grass (Poa pratensis L. and white clover (Trifolium repens L. in hill mountain areas. In a three-year average, the application of 150 kg ha-1 N had significant impact on cocksfoot population density (number of tillers m-2, and it was 51.4 % higher than the recorded one before grazing (P<0.05 and 42.2 % higher after grazing (P<0.01 in comparison to N0. The application of 150 kg ha-1 N resulted in significantly higher cocksfoot dry matter (DM yield for 38.6 % (P<0.01 and 15.3 % higher cocksfoot share in the total mixture in relation to N0 (P<0.01. Grazing management and grazing management interaction with N rate did not significantly affect the population density of individual years. However, in the three-year average, grazing management significantly affected cocksfoot DM (P<0.01 and its percentage in the total DM mixture (P<0.01. Cattle grazing resulted in 9.9 % higher cocksfoot DM yield and 15.2 % higher cocksfoot percentage in pasture. Interaction of grazing management and N-level had significant influence on the percentage of cocksfoot DM in grass-clover mixture. On cattle grazed areas fertilized with 150 kg ha-1 N, the percentage of cocksfoot DM was the highest (74. 07%, while the lowest percentage of cocksfoot DM was recorded on the sheep grazed areas without N (55.12%.

  18. Kunitz Proteinase Inhibitors Limit Water Stress Responses in White Clover (Trifolium repens L. Plants

    Directory of Open Access Journals (Sweden)

    Afsana Islam

    2017-10-01

    Full Text Available The response of plants to water deficiency or drought is a complex process, the perception of which is triggered at the molecular level before any visible morphological responses are detected. It was found that different groups of plant proteinase inhibitors (PIs are induced and play an active role during abiotic stress conditions such as drought. Our previous work with the white clover (Trifolium repens L. Kunitz Proteinase Inhibitor (Tr-KPI gene family showed that Tr-KPIs are differentially regulated to ontogenetic and biotic stress associated cues and that, at least some members of this gene family may be required to maintain cellular homeostasis. Altered cellular homeostasis may also affect abiotic stress responses and therefore, we aimed to understand if distinct Tr-PKI members function during drought stress. First, the expression level of three Tr-KPI genes, Tr-KPI1, Tr-KPI2, and Tr-KPI5, was measured in two cultivars and one white clover ecotype with differing capacity to tolerate drought. The expression of Tr-KPI1 and Tr-KPI5 increased in response to water deficiency and this was exaggerated when the plants were treated with a previous period of water deficiency. In contrast, proline accumulation and increased expression of Tr-NCED1, a gene encoding a protein involved in ABA biosynthesis, was delayed in plants that experienced a previous drought period. RNAi knock-down of Tr-KPI1 and Tr-KPI5 resulted in increased proline accumulation in leaf tissue of plants grown under both well-watered and water-deficit conditions. In addition, increased expression of genes involved in ethylene biosynthesis was found. The data suggests that Tr-KPIs, particularly Tr-KPI5, have an explicit function during water limitation. The results also imply that the Tr-KPI family has different in planta proteinase targets and that the functions of this protein family are not solely restricted to one of storage proteins or in response to biotic stress.

  19. Functional analysis and localisation of a delta-class glutathione S-transferase from Sarcoptes scabiei.

    Science.gov (United States)

    Pettersson, Eva U; Ljunggren, Erland L; Morrison, David A; Mattsson, Jens G

    2005-01-01

    The mite Sarcoptes scabiei causes sarcoptic mange, or scabies, a disease that affects both animals and humans worldwide. Our interest in S. scabiei led us to further characterise a glutathione S-transferase. This multifunctional enzyme is a target for vaccine and drug development in several parasitic diseases. The S. scabiei glutathione S-transferase open reading frame reported here is 684 nucleotides long and yields a protein with a predicted molecular mass of 26 kDa. Through phylogenetic analysis the enzyme was classified as a delta-class glutathione S-transferase, and our paper is the first to report that delta-class glutathione S-transferases occur in organisms other than insects. The recombinant S. scabiei glutathione S-transferase was expressed in Escherichia coli via three different constructs and purified for biochemical analysis. The S. scabiei glutathione S-transferase was active towards the substrate 1-chloro-2,4-dinitrobenzene, though the positioning of fusion partners influenced the kinetic activity of the enzyme. Polyclonal antibodies raised against S. scabiei glutathione S-transferase specifically localised the enzyme to the integument of the epidermis and cavities surrounding internal organs in adult parasites. However, some minor staining of parasite intestines was observed. No staining was seen in host tissues, nor could we detect any antibody response against S. scabiei glutathione S-transferase in sera from naturally S. scabiei infected dogs or pigs. Additionally, the polyclonal sera raised against recombinant S. scabiei glutathione S-transferase readily detected a protein from mites, corresponding to the predicted size of native glutathione S-transferase.

  20. Polish Yellow Sweet Clover (Melilotus officinalis L. Honey, Chromatographic Fingerprints, and Chemical Markers

    Directory of Open Access Journals (Sweden)

    Izabela Jasicka-Misiak

    2017-01-01

    Full Text Available A case study of Polish Melilotus officinalis honey was presented for the first time. Gas chromatography–mass spectrometry (GC-MS (after steam distillation, Soxhlet extraction, ultrasonic solvent extraction, and solid phase extraction (SPE and targeted high performance liquid chromatography with a photodiode array detector (HPLC-PAD were applied to determine the characteristic components of honey. While ubiquitous in most honeys, carbohydrates, terpene derivatives, and phenylacetic acid dominated in the Soxhlet extracts (25.54% and in the application of SPE (13.04%. In addition, lumichrome (1.85% was found, and may be considered as a marker of this honey. Due to the presence of these compounds, Polish yellow sweet clover honey is similar to French lavender honeys. The major compounds determined in the methanolic extract were (+-catechine (39.7% and gallic acid (up to 30%, which can be regarded as specific chemical markers of the botanical origin of melilot honey. With respect to total phenolic and flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl (DPPH assays were determined spectrophotometrically. The honey exhibited a moderate antioxidant activity, typical for light honeys, which correlates well with its phenolic and flavonoid composition.

  1. Proportional crosstalk correction for the segmented clover at iThemba LABS

    International Nuclear Information System (INIS)

    Bucher, T D; Noncolela, S P; Lawrie, E A; Dinoko, T R S; Easton, J L; Erasmus, N; Lawrie, J J; Mthembu, S H; Mtshali, W X; Shirinda, O; Orce, J N

    2017-01-01

    Reaching new depths in nuclear structure investigations requires new experimental equipment and new techniques of data analysis. The modern γ -ray spectrometers, like AGATA and GRETINA are now built of new-generation segmented germanium detectors. These most advanced detectors are able to reconstruct the trajectory of a γ -ray inside the detector. These are powerful detectors, but they need careful characterization, since their output signals are more complex. For instance for each γ -ray interaction that occurs in a segment of such a detector additional output signals (called proportional crosstalk), falsely appearing as an independent (often negative) energy depositions, are registered on the non-interacting segments. A failure to implement crosstalk correction results in incorrectly measured energies on the segments for two- and higher-fold events. It affects all experiments which rely on the recorded segment energies. Furthermore incorrectly recorded energies on the segments cause a failure to reconstruct the γ -ray trajectories using Compton scattering analysis. The proportional crosstalk for the iThemba LABS segmented clover was measured and a crosstalk correction was successfully implemented. The measured crosstalk-corrected energies show good agreement with the true γ -ray energies independent on the number of hit segments and an improved energy resolution for the segment sum energy was obtained. (paper)

  2. Effects of Seeding Density and Nitrogen Fertilizer on the Productivity of Egyptian Clover

    Directory of Open Access Journals (Sweden)

    Jwan G. Rafaat

    2013-11-01

    Full Text Available The study was carried out to show the effect of different levels of nitrogen fertilizer 0, 20, 40 and 60 kg urea/ha, and two seeding rates 15 and 30 kg/ha. The study was conducted at Bakrajo research field during the winter season 2011-2012 to some growth characteristics of Egyptian clover, such as plant height, dry leaf weight percent, dry stem weight percent, leave stem ratio, fresh yield t/ha, dry yield t/ha and dry matter percent. The experiment was designed as (R.C.B.D. The results can be summarized as follow; significant differences were observed between all three cuts, and the third cut was superior in almost characters especially in the forage yield. The application of 40 and 60 kg urea gave maximum yield. Using 15 kg seeds/ha showed superior value due to fresh yield in compare to 30 kg for all cutting, while the dry yield responded non-significantly to seeding rates.

  3. Effect of forage supplements on the incidence of bloat in dairy cows grazing high clover pastures.

    Science.gov (United States)

    Phillips, C J; James, N L; Murray-Evans, J P

    1996-08-17

    The effect of offering forage supplements of different compositions was examined in two experiments with cows grazing high clover swards. In the first experiment strawmix supplements of high or low energy content (11 and 9 MJ metabolisable energy/kg dry matter [DM]) and high or low crude protein content (17 and 4 g/kg DM) were offered for periods of three weeks. The energy and protein contents were varied by the content of molasses and soyabean meal, respectively. The high energy, high protein supplement increased the incidence of bloat, and the low energy, high protein supplement reduced it, compared with grazing alone. Bloat was most evident in the first two weeks of each feeding period, suggesting that the cows partially adapted to the diets within three weeks. In the second experiment silage supplements reduced the incidence of bloat among cows grazing both tall and short swards. The most suitable forages to feed when there is a risk of bloat are those that are slowly fermented in the rumen but are eaten in sufficient quantity to reduce periods of rapid herbage intake.

  4. Fecal microbiota of lambs fed purple prairie clover (Dalea purpurea Vent.) and alfalfa (Medicago sativa).

    Science.gov (United States)

    Huang, Qianqian; Holman, Devin B; Alexander, Trevor; Hu, Tianming; Jin, Long; Xu, Zhongjun; McAllister, Tim A; Acharya, Surya; Zhao, Guoqi; Wang, Yuxi

    2018-01-01

    The present study assessed the effect of purple prairie clover (PPC) and PPC condensed tannins (CT) on the fecal microbiota of lambs using high-throughput 16S rRNA gene pyrosequencing. A total of 18 individual lambs were randomly divided into three groups and fed either green chop alfalfa (Alf), a 40:60 (DM basis; Mix) mixture of Alf and PPC, or Mix supplemented with polyethylene glycol (Mix-P) for 18 days. Fecal samples were collected on days 13 through 18 using digital rectal retrieval. The DNA of fecal samples was extracted and the microbial 16S rRNA gene amplicons were sequenced using 454 pyrosequencing. Regardless of diet, the bacterial community was dominated by Firmicutes and Bacteroidetes with many sequences unclassified at the genus level. Forage type and CT had no effect on the fecal microbial composition at the phylum level or on α-diversity. Compared to the Alf diet, the Mix diet reduced the relative abundance of Akkermansia (P = 0.03) and Asteroleplasma (P = 0.05). Fecal microbial populations in Alf and Mix-P clustered separately from each other when assessed using unweighted UniFrac (P < 0.05). These results indicate that PPC CT up to 36 g/kg DM in the diet had no major effect on fecal microbial flora at the phyla level and exerted only minor effects on the genera composition of fecal microbiota in lambs.

  5. Genetic variability of isoflavones in the USDA red clover core collection

    Directory of Open Access Journals (Sweden)

    Graziele P. Ramos

    2012-11-01

    Full Text Available Red clover is one of the most utilized forage in agriculture and contains many of the isoflavones known for their human health benefits. The objectives of this study were: i to quantify, using HPLC analysis, isoflavones in 77 accessions from the USDA core collection and a Brazilian line; ii to verify possible relationships depending on their origin, improvement status or maturity type and; iii to verify the seasonal variation. The isoflavone mean contents were 29.27 µg g-1 of dry material for daidzein, 163.69 µg g-1 for genistein, 11353.29 µg g-1 for formononetin and 6568.8 µg g-1 for biochanin A. Clustering was mainly influenced by the total amount of isoflavones and partially due to maturity type, improvement status and geographic origin. The seasonal evaluation demonstrated an increase of concentration during winter, and decrease during spring. These results highlighted accessions that can be used to develop new varieties with low or high isoflavones concentration.

  6. A study of the red clover extract trinovin by ESR HPLC/MS and UVS

    International Nuclear Information System (INIS)

    Troup, G.; Hutton, D.; Hunter, C.; Hewitt, D.; Mulinacci, N.; Romani, A.; Pinelli, P.; Mancini, P.

    1999-01-01

    Full text: Trinovin is an extract of red clover, recently released on the dietary supplement market. It is recommended for 'Men's Health', because it contains the phenolics (isoflavones) genistein, biochanin, daidzein and formononetin, said to act as 'phytoestrogens', and is therefore a possible help in prostate gland problems. An Electron Spin Resonance (ESR) study (∼9.1Ghz, room temperature) revealed at least 3 different free radical lines, one with hyperfine structure, consistent with the listed molecules. Accordingly, HPLC/DAD (High Performance Liquid Chromatography/Diode Array Detector) and HPLC/Mass Spectroscopy analyses were performed in order to evaluate the quali-quantitative contents of flavonoidic compounds. The HPLC profile shows two main isoflavones and another three compounds, one of them being a quercetin glycoside. The quercetin glycosides are flavonoidic derivatives abundant in plant materials and present in wine. We can therefore say: even if the phytoestrogen properties claimed for Trinovin turn out to be less than hoped for, the antioxidants contained are very powerful, and so possibly helpful in protection against many diseases, including cancers, atherosclerosis, diabetic retinal bleeding, and non-alcoholic dementia

  7. In vitro isoflavonoid production and analysis in natural tetraploid Trifolium pratense (red clover calluses

    Directory of Open Access Journals (Sweden)

    Tugba Ercetin

    2012-09-01

    Full Text Available Isoflavones are polyphenolic phytoestrogens, predominantly found in leguminous plants. Trifolium pratense L., Fabaceae (red clover, is rich in isoflavones that possess estrogenic activity due to their similar molecular structure and effectiveness in preventing health conditions such as menopause, osteoporosis, cardiovascular disease, hypertension and hormone-dependent cancers. In this study, presence and amount of various phytoestrogens in the tetraploid plant and in the calluses derived from the plants were investigated. Calluses were generated from explants obtained from natural tetraploid T. pratense seedlings. The best callus formation was obtained from hypocotyl explants cultured in Phillips Collins and Gamborg B5 media containing different plant growth regulators. Flowers of plants and calluses were analysed for formononetin, biochanin A, genistein and daidzein contents using HPLC. In HPLC analysis, high levels of formononetin (0.249 µg/mg were determined in natural tetraploid T. pratense flowers in addition to genistein and biochanin A. In calluses, highest isoflavone content (1.15 µg/mg formononetin was observed in modified Gamborg B5 medium. Biochanin A content of calluses and the plant were found to be nearly the same. But formononetin and genistein contents of the calluses in this medium were found to be respectively 4.62 and 21.39 folds higher than the tetraploid plant.

  8. In vitro isoflavonoid production and analysis in natural tetraploid Trifolium pratense (red clover calluses

    Directory of Open Access Journals (Sweden)

    Tugba Ercetin

    2012-10-01

    Full Text Available Isoflavones are polyphenolic phytoestrogens, predominantly found in leguminous plants. Trifolium pratense L., Fabaceae (red clover, is rich in isoflavones that possess estrogenic activity due to their similar molecular structure and effectiveness in preventing health conditions such as menopause, osteoporosis, cardiovascular disease, hypertension and hormone-dependent cancers. In this study, presence and amount of various phytoestrogens in the tetraploid plant and in the calluses derived from the plants were investigated. Calluses were generated from explants obtained from natural tetraploid T. pratense seedlings. The best callus formation was obtained from hypocotyl explants cultured in Phillips Collins and Gamborg B5 media containing different plant growth regulators. Flowers of plants and calluses were analysed for formononetin, biochanin A, genistein and daidzein contents using HPLC. In HPLC analysis, high levels of formononetin (0.249 µg/mg were determined in natural tetraploid T. pratense flowers in addition to genistein and biochanin A. In calluses, highest isoflavone content (1.15 µg/mg formononetin was observed in modified Gamborg B5 medium. Biochanin A content of calluses and the plant were found to be nearly the same. But formononetin and genistein contents of the calluses in this medium were found to be respectively 4.62 and 21.39 folds higher than the tetraploid plant.

  9. Mitigation of salt stress in white clover (Trifolium repens) by Azospirillum brasilense and its inoculation effect.

    Science.gov (United States)

    Khalid, Muhammad; Bilal, Muhammad; Hassani, Danial; Iqbal, Hafiz M N; Wang, Hang; Huang, Danfeng

    2017-12-01

    Salinity is one of the increasingly serious environmental problems worldwide for cultivating agricultural crops. The present study was aimed to ascertain the potential of beneficial soil bacterium Azospirillum brasilense to alleviate saline stress in Trifolium repens. Experimental plants (white clover) were grown from seeds and inoculated with or without A. brasilense bacterial strain supplemented with 0, 40, 80, or 120 mM NaCl into soil. The growth attributes including, shoot heights, root lengths, fresh and dry weights, leaf area and chlorophyll content were significantly enhanced in T. repens plants grown in A. brasilense inoculated soil than un-inoculated controls, particularly under elevated salinity conditions (40, 80 and 120 mM NaCl). Malondialdehyde content of leaf was recorded to be declined under saline conditions. Moreover, the K + /Na + ratio was also improved in bacterium-inoculated plants, since A. brasilense significantly reduced the root and shoot Na + level under high salty environment. Results revealed that soil inoculation with A. brasilense could significantly promote T. repens growth under both non-saline and saline environments, and this study might be extended to other vegetables and crops for the germination and growth enhancement.

  10. The role of glutathione transferases in renal cell carcinoma

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    Ćorić Vesna

    2016-01-01

    Full Text Available Mounting evidence suggest that members of the subfamily of cytosolic glutathione S-transferases (GSTs possess roles far beyond the classical glutathione-dependent enzymatic conjugation of electrophilic metabolites and xenobiotics. Namely, monomeric forms of certain GSTs are capable of forming protein: protein interactions with protein kinases and regulate cell apoptotic pathways. Due to this dual functionality of cytosolic GSTs, they might be implicated in both the development and the progression of renal cell carcinoma (RCC. Prominent genetic heterogeneity, resulting from the gene deletions, as well as from SNPs in the coding and non-coding regions of GST genes, might affect GST isoenzyme profiles in renal parenchyma and therefore serve as a valuable indicator for predicting the risk of cancer development. Namely, GSTs are involved in the biotransformation of several compounds recognized as risk factors for RCC. The most potent carcinogen of polycyclic aromatic hydrocarbon diol epoxides, present in cigarette smoke, is of benzo(apyrene (BPDE, detoxified by GSTs. So far, the relationship between GST genotype and BPDE-DNA adduct formation, in determining the risk for RCC, has not been evaluated in patients with RCC. Although the association between certain individual and combined GST genotypes and RCC risk has been debated in a the literature, the data on the prognostic value of GST polymorphism in patients with RCC are scarce, probably due to the fact that the molecular mechanism supporting the role of GSTs in RCC progression has not been clarified as yet.

  11. Glutathione transferase-mediated benzimidazole-resistance in Fusarium graminearum.

    Science.gov (United States)

    Sevastos, A; Labrou, N E; Flouri, F; Malandrakis, A

    2017-09-01

    Fusarium graminearum laboratory mutants moderately (MR) and highly (HR) benzimidazole-resistant, carrying or not target-site mutations at the β 2 -tubulin gene were utilized in an attempt to elucidate the biochemical mechanism(s) underlying the unique BZM-resistance paradigm of this fungal plant pathogen. Relative expression analysis in the presence or absence of carbendazim (methyl-2-benzimidazole carbamate) using a quantitative Real Time qPCR (RT-qPCR) revealed differences between resistant and the wild-type parental strain although no differences in expression levels of either β 1 - or β 2 -tubulin homologue genes were able to fully account for two of the highly resistant phenotypes. Glutathione transferase (GST)-mediated detoxification was shown to be -at least partly- responsible for the elevated resistance levels of a HR isolate bearing the β 2 -tubulin Phe200Tyr resistance mutation compared with another MR isolate carrying the same mutation. This benzimidazole-resistance mechanism is reported for the first time in F. graminearum. No indications of detoxification involved in benzimidazole resistance were found for the rest of the isolates as revealed by GST and glutathione peroxidase (GPx) activities and bioassays using monoxygenase and hydrolase detoxification enzyme inhibiting synergists. Interestingly, besides the Phe200Tyr mutation-carrying HR isolate, the remaining highly-carbendazim resistant phenotypes could not be associated with any of the target site modification/overproduction, detoxification or reduced uptake-increased efflux mechanisms. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Glutathione Transferase GSTπ In Breast Tumors Evaluated By Three Techniques

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    Rafael Molina

    1993-01-01

    Full Text Available The glutathione transferases are involved in intracellular detoxification reactions. One of these, GSTπ, is elevated in some breast cancer cells, particularly cells selected for resistance to anticancer agents. We evaluated GSTπ expression in 60 human breast tumors by three techniques, immunohistochemistry, Northern hybridization, and Western blot analysis. There was a significant positive correlation between the three methods, with complete concordance seen in 64% of the tumors. There was strong, inverse relationship between GSTπ expression and steroid receptor status with all of the techniques utili zed. [n addition, there was a trend toward higher GSTπ expression in poorly differentiated tumors, but no correlation was found between tumor GSTπ content and DNA ploidy or %S-phase. GSTπ expression was also detected in adjacent benign breast tissue as well as infiltrating lymphocytes; this expression may contribute to GSTπ measurements using either Northern hybridization or Western blot analysis. These re sults suggest that immunohistochemistry is the method of choice for measuring GSTπ in breast tumors.

  13. Glycoproteins and sialyl transferase of human B lymphoblastoid cell lines

    International Nuclear Information System (INIS)

    Lui, S.W.L.; Ng, M.H.

    1980-01-01

    We used two radiolabeling methods to study glycoproteins on the surface of lymphoblastoid cells. One of the methods affects tritiation of residues which are oxidized with galactose oxidase and the other causes tritiation of neuraminic acid residues. This approach was shown to allow a better resolution of cell surface glycoproteins than if either method were used alone. Glycoproteins of B 1 - 19 cells which harbor the Epstein-Barr virus genomes were compared with those of its parental cell line, BJAB, which does not harbor the viral genomes. These studies did not reveal a unique viral protein. A 28,000 mol. wt. glycoprotein was found to be the most prominent neuraminic acidlabeled product of B 1 - 19 cells and also of the two other cell lines, Raji and Ly38, which harbor the EBV genomes. A similar molecular weight species from BJAB cells identified by galactose oxidase labeling might be deficient in neuraminic acid residues as it was poorly labeled by the periodate oxidation method. The neuraminic acid content and level of sialyl transferase of BJAB cells were found to be lower than those of the other cell lines studied. (auth.)

  14. S-Nitrosation destabilizes glutathione transferase P1-1.

    Science.gov (United States)

    Balchin, David; Stoychev, Stoyan H; Dirr, Heini W

    2013-12-23

    Protein S-nitrosation is a post-translational modification that regulates the function of more than 500 human proteins. Despite its apparent physiological significance, S-nitrosation is poorly understood at a molecular level. Here, we investigated the effect of S-nitrosation on the activity, structure, stability, and dynamics of human glutathione transferase P1-1 (GSTP1-1), an important detoxification enzyme ubiquitous in aerobes. S-Nitrosation at Cys47 and Cys101 reduces the activity of the enzyme by 94%. Circular dichroism spectroscopy, acrylamide quenching, and amide hydrogen-deuterium exchange mass spectrometry experiments indicate that the loss of activity is caused by the introduction of local disorder at the active site of GSTP1-1. Furthermore, the modification destabilizes domain 1 of GSTP1-1 against denaturation, smoothing the unfolding energy landscape of the protein and introducing a refolding defect. In contrast, S-nitrosation at Cys101 alone introduces a refolding defect in domain 1 but compensates by stabilizing the domain kinetically. These data elucidate the physical basis for the regulation of GSTP1-1 by S-nitrosation and provide general insight into the consequences of S-nitrosation on protein stability and dynamics.

  15. Biologically fixed N2 as a source for N2O production in a grass–clover mixture, measured by 15N2 (erratum i vol. 74 p. 203)

    DEFF Research Database (Denmark)

    Carter, Mette Sustmann; Ambus, P.

    2006-01-01

    The contribution of biologically fixed dinitrogen (N-2) to the nitrous oxide (N2O) production in grasslands is unknown. To assess the contribution of recently fixed N-2 as a source of N2O and the transfer of fixed N from clover to companion grass, mixtures of white clover and perennial ryegrass...

  16. High Statistics Analysis using Anisotropic Clover Lattices: (IV) The Volume Dependence of the Light Hadron Masses

    Energy Technology Data Exchange (ETDEWEB)

    Beane, S R; Detmold, W; Lin, H W; Luu, T C; Orginos, K; Parreno, A; Savage, M J; Torok, A; Walker-Loud, A

    2011-07-01

    The volume dependence of the octet baryon masses and relations among them are explored with Lattice QCD. Calculations are performed with nf = 2 + 1 clover fermion discretization in four lattice volumes, with spatial extent L ? 2.0, 2.5, 3.0 and 4.0 fm, with an anisotropic lattice spacing of b_s ? 0.123 fm in the spatial direction, and b_t = b_s/3.5 in the time direction, and at a pion mass of m_\\pi ? 390 MeV. The typical precision of the ground-state baryon mass determination is

  17. Climate Clever Clovers: New Paradigm to Reduce the Environmental Footprint of Ruminants by Breeding Low Methanogenic Forages Utilizing Haplotype Variation

    Directory of Open Access Journals (Sweden)

    Parwinder Kaur

    2017-09-01

    Full Text Available Mitigating methane production by ruminants is a significant challenge to global livestock production. This research offers a new paradigm to reduce methane emissions from ruminants by breeding climate-clever clovers. We demonstrate wide genetic diversity for the trait methanogenic potential in Australia’s key pasture legume, subterranean clover (Trifolium subterraneum L.. In a bi-parental population the broadsense heritability in methanogenic potential was moderate (H2 = 0.4 and allelic variation in a region of Chr 8 accounted for 7.8% of phenotypic variation. In a genome-wide association study we identified four loci controlling methanogenic potential assessed by an in vitro fermentation system. Significantly, the discovery of a single nucleotide polymorphism (SNP on Chr 5 in a defined haplotype block with an upstream putative candidate gene from a plant peroxidase-like superfamily (TSub_g18548 and a downstream lectin receptor protein kinase (TSub_g18549 provides valuable candidates for an assay for this complex trait. In this way haplotype variation can be tracked to breed pastures with reduced methanogenic potential. Of the quantitative trait loci candidates, the DNA-damage-repair/toleration DRT100-like protein (TSub_g26967, linked to avoid the severity of DNA damage induced by secondary metabolites, is considered central to enteric methane production, as are disease resistance (TSub_g26971, TSub_g26972, and TSub_g18549 and ribonuclease proteins (TSub_g26974, TSub_g26975. These proteins are good pointers to elucidate the genetic basis of in vitro microbial fermentability and enteric methanogenic potential in subterranean clover. The genes identified allow the design of a suite of markers for marker-assisted selection to reduce rumen methane emission in selected pasture legumes. We demonstrate the feasibility of a plant breeding approach without compromising animal productivity to mitigate enteric methane emissions, which is one of the most

  18. The clover technique for the treatment of complex tricuspid valve insufficiency: midterm clinical and echocardiographic results in 66 patients.

    Science.gov (United States)

    Lapenna, Elisabetta; De Bonis, Michele; Verzini, Alessandro; La Canna, Giovanni; Ferrara, David; Calabrese, Maria Chiara; Taramasso, Maurizio; Alfieri, Ottavio

    2010-06-01

    This study assesses the results of the 'clover technique' (suturing together the middle point of the free edges of the tricuspid leaflets) for the treatment of tricuspid regurgitation (TR) due to severe prolapse or tethering. From 2001, 66 patients with severe TR due to prolapsing or tethered leaflets underwent 'clover repair'. Annuloplasty was associated in 64 patients (97%). The aetiology of TR was degenerative in 52 cases (79%), post-traumatic in eight (12%) and secondary to dilated cardiomyopathy (DCM) in six (9%). The main mechanism of TR was prolapse/flail of one leaflet in 15 patients (23%), of two leaflets in 31 (47%) and of all three leaflets in 14 (21%). The remaining six patients (9%) presented with severe leaflets' tethering. Four deaths (6%) occurred during hospitalisation and one patient died 3.6 years after surgery. Survival was 91 + or - 4.1% at 5 years. Follow-up of the 62 hospital survivors was 100% complete (mean length 3.5 + or - 1.6 years, range 13 months-7.1 years). At the last echocardiogram, no or mild TR was detected in 55 (88.7%) patients, moderate (2+/4+) in six (9.6%) and severe (4+/4+) in one patient (1.6%). Mean tricuspid valve area and gradient were 4.3 + or - 0.6 cm(2) and 2.8 + or -1.4 mmHg. In six patients, stress echocardiography was performed and no signs of tricuspid stenosis were detected. At the multivariable analysis, the degree of TR at hospital discharge was identified as the only predictor of TR > or = 2+ at follow-up. Midterm clinical and echocardiographic results confirm the role of the 'clover technique' in the surgical treatment of TR due to lesions, which are unlikely to be effectively treatable by annuloplasty alone. Copyright 2010 European Association for Cardio-Thoracic Surgery. Published by Elsevier B.V. All rights reserved.

  19. The Effects of Ensiled Berseem Clover and Citrus Pulp Mixture on Performance of Zel Fattened Lambs

    Directory of Open Access Journals (Sweden)

    maedeh feyz

    2016-08-01

    Full Text Available Introduction Feed contributes about 75% of the total cost of animal production, therefore utilizing of by-products such as Berseem clover and citrus pulp, as nutritive and low cost components of rations would decrease the production cost. In north of Iran over autumn and winter, utilizing of these by-products in making of silage as feed for ruminants provides good feed ingredient especially in feedlot operations, also eliminates pathogens, and reduces the effect of drugs and pesticides that are used locally without a serious control or discipline. However, little information available on utilizing silage made from these local by-products. The objectives of this research were to investigate the effects of ensiled Berseem clover and orange peels mixture on intake, digestibility, chewing behavior and performance of Zel fattening lambs. Materials and methods Twenty male Zel lambs fed with five experimental rations containing basal concentrate and 35% Berseem clover silage as: 1 without additives, 2 supplemented with 40% dried orange peels, 3 supplemented with 40% dried tangerine peel, 4 supplemented with 35% dried tangerine peel and 5% ground barley and 5 supplemented with 35% dried orange peels and 5% ground barley. Lambs were housed in individual box and fed ad libitum, twice daily at 09:00 and 21:00 h with total mixed rations as experimental treatments, allowing for at least 10% residuals (as-fed basis. Water and mineralized salt stone were available throughout the experiment. Feed particle size distribution, geometric mean and the standard deviation of geometric mean were determined by dry sieving in four replicates, using two set of Penn State particle separator. Feed, feces and orts were analyzed for dry matter, Kjeldahl N, ether extract, organic matter and ash at 605°C, neutral and acid detergent fiber (NDF and ADF when α-amylase being added for concentrates during NDF extraction; sodium sulfite was not added. Neutral detergent fiber was

  20. Acetate:succinate CoA-transferase in the hydrogenosomes of Trichomonas vaginalis: Identification and characterization

    NARCIS (Netherlands)

    K.W.A. Grinsven; S. Rosnowsky (Silke); S.W.H. van Weelden (Susanne); S. Pütz (Simone); M. van der Giezen (Mark); W. Martin (William); J.J. van Hellemond (Jaap); A.G.M. Tielens (Aloysius); K. Henze (Katrin)

    2008-01-01

    textabstractAcetate:succinate CoA-transferases (ASCT) are acetate-producing enzymes in hydrogenosomes, anaerobically functioning mitochondria and in the aerobically functioning mitochondria of trypanosomatids. Although acetate is produced in the hydrogenosomes of a number of anaerobic microbial

  1. Hydrographs Showing Groundwater Level Changes for Selected Wells in the Chambers-Clover Creek Watershed and Vicinity, Pierce County, Washington

    Science.gov (United States)

    Justin, G.B.; Julich, R.; Payne, K.L.

    2009-01-01

    Selected groundwater level hydrographs for the Chambers-Clover Creek watershed (CCCW) and vicinity, Washington, are presented in an interactive web-based map to illustrate changes in groundwater levels in and near the CCCW on a monthly and seasonal basis. Hydrographs are linked to points corresponding to the well location on an interactive map of the study area. Groundwater level data and well information from Federal, State, and local agencies were obtained from the U.S. Geological Survey National Water Information System (NWIS), Groundwater Site Inventory (GWSI) System.

  2. Distribution of radionuclides in leaf-stem biomass of lupine and clover under production of protein concentrates

    International Nuclear Information System (INIS)

    Novikov, Yu.F.; Lobach, G.A.; Buzenko, T.A.; Zaretskaya, T.P.

    1993-01-01

    The basic regularities of radionuclide distribution between the obtained products have been studied using the fractionation of lupine and clover phytomass as an example. The content of radionuclides in protein concentrates has been shown to be strongly related to the crop species. A scheme and a regime of the fractionation of leaf-stem lupine biomass contaminated with cesium radioisotopes and strontium-90 which ensured the minimizing of their residual content in protein-vitaminic and protein concentrates have been selected with due accout of experimental data

  3. Perturbative determination of c{sub sw} for plaquette and Symanzik gauge action and stout link clover fermions

    Energy Technology Data Exchange (ETDEWEB)

    Horsley, R. [Edinburgh Univ. (United Kingdom). School of Physics; Perlt, H.; Schiller, A. [Leipzig Univ. (Germany). Inst. fuer Theoretische Physik; Rakow, P.E.L. [Liverpool Univ. (United Kingdom). Theoretical Physics Division, Dept. of Mathematical Sicences; Schierholz, G. [Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)

    2008-06-15

    Using plaquette and Symanzik improved gauge action and stout link clover fermions we determine the improvement coefficient c{sub SW} in one-loop lattice perturbation theory from the off-shell quark-quark-gluon three-point function. In addition, we compute the coefficients needed for the most general form of quark field improvement and present the one-loop result for the critical hopping parameter {kappa}{sub c}. We discuss mean field improvement for c{sub SW} and {kappa}{sub c} and the choice of the mean field coupling for the actions we have considered. (orig.)

  4. Fatty acid composition of ruminal digesta and longissimus muscle from lambs fed silage mixtures including red clover, sainfoin, and timothy.

    Science.gov (United States)

    Campidonico, L; Toral, P G; Priolo, A; Luciano, G; Valenti, B; Hervás, G; Frutos, P; Copani, G; Ginane, C; Niderkorn, V

    2016-04-01

    This work investigated the effects of feeding silage mixtures of a plant containing polyphenol oxidase (PPO; red clover [; RC]), a plant containing tannins (sainfoin [; SF]), and a grass species not containing these compounds (timothy [; T]) on ruminal and intramuscular (i.m.) fatty acids of lambs. Forty 4-mo-old castrated male Romane lambs, divided into 5 groups, received 1 of the following silages: 1) T (100%), 2) a binary mixture of timothy and tannin-containing sainfoin ( cv. Perly; 50:50 [T-SF]), 3) a binary mixture of timothy and PPO-containing red clover ( cv. Mervius; 50:50 [T-RC]), 4) a ternary mixture of timothy, sainfoin, and red clover containing both tannins and PPO (50:25:25, respectively [T-SF-RC]), and 5) a binary mixture of tannin-containing sainfoin and PPO-containing red clover (50:50 [SF-RC]). In the rumen digesta, the partial or total replacement of T with forage legumes was associated with greater concentrations of PUFA ( forage legumes in the silage favored the accumulation of 18:3 -3 ( Forage legumes decreased the -11 18:1 to 30% of T in rumen digesta ( Forage legumes decreased the total concentration of branched-chain fatty acids in the rumen digesta (on average, -28%; comparison with T. The dietary treatment tended to affect the proportion of MUFA ( = 0.081) and of PUFA ( = 0.079) in the i.m. fat of the LM, respectively, at the highest and lowest numerical value in the T group. The sum of -3 fatty acids was less in the T and T-SF groups compared with the mixture of legumes without T (SF-RC; comparison with lambs given T-RC, T-SF-RC, and SF-RC. Rumenic acid (-9 -11 CLA) was detected at a greater percentage in the LM from the animals fed the T silage compared with animals fed the T-SF-RC treatment ( = 0.004). Contrarily, -9 -12 -15 18:3 was found at a greater concentration in the muscle from lambs in the SF-RC treatment compared with lambs in the other treatments ( forage for growing lambs in terms of i.m. fatty acid composition.

  5. Design and fabrication of 4π Clover Detector Array Assembly for gamma-spectroscopy studies using thermal neutrons

    International Nuclear Information System (INIS)

    Kumar, Manish; Kamble, S.R.; Chaudhari, A.T.; Sabharwal, T.P.; Pathak, Kavindra; Prasad, N.K.; Kinage, L.A.; Biswas, D.C.; Bhagwat, P.V.

    2017-01-01

    Nuclear spectroscopy has been studied earlier from the measurement of prompt gamma rays produced in reactions with thermal neutrons from CIRUS reactor. For studying the prompt γ-spectroscopy using thermal neutrons from Dhruva Reactor, BARC, the development of a dedicated beam line (R-3001) is in progress. In this beam line a detector assembly consisting of Clover Ge detectors will be used. This experimental setup will be utilized to investigate nuclear structure using prompt (n,γ) reactions and also to study the spectroscopy of neutron-rich fission-fragment nuclei

  6. Effect of Plant Density on Growth Characteristics and Yield of Summer Savory (Satureja hortensis L. and Persian Clover (Trifolium resupinatum L. Intercropping

    Directory of Open Access Journals (Sweden)

    f Hassanzadeh Aval

    2012-02-01

    Full Text Available Abstract In order to evaluate intercropping of summer savory (Satureja hortensis L. and Persian clover (Trifolium resupinatum L., an experiment was conducted in the Agricultural Research Station of Ferdowsi University of Mashhad in 2004 growing season. Treatments were sole cropping of Persian clover (eight rows, double-row intercropping of Persian clover and summer savory with 27, 40 and 80 plants.m-2 (eight rows and sole cropping of summer savory with 27, 40 and 80 plants m-2 (eight rows. For this purpose a complete randomized block design with 4 replications was used. Shoot and stem percentage of summer savory in sole crop treatments were significantly higher than in intercrop. In sole crop treatments, these parameters decreased by increasing plant density, in contrast to the intercrop. Leaf percentage and leaf and flower to stem ratio of summer savory in sole crop treatments were significantly lower than in intercrop. In sole crop treatments, these parameters were increased by increasing plant density, in contrast to intercrop. Effect of different treatments on essential oil percentage of summer savory was not significant. In sole cropping of Persian clover treatment, dry weight of vegetative organs and stem percentage of Persian clover in the first harvest, was lower than other treatments. By decreasing plant density these parameters were decreased in intercropping. In the second and third harvests a reversed pattern was observed. The highest Area Time Equivalent Ratio was obtained in intercropping of persian clover and summer savory with 27 plants.m-2. Keywords: Intercropping, Plant density, Satureja hortensis, Trifolium resupinatum, Essential oil percentage, Area Time Equivalent Ratio

  7. Estimating legume N-2 fixation in grass-clover mixtures of a grazed organic cropping system using two N-15 methods

    DEFF Research Database (Denmark)

    Vinther, F.P.; Jensen, E.S.

    2000-01-01

    The input of Nitrogen (N) through symbiotic N-2 fixation (SNF) in grass-clover mixtures was determined in an organic cropping. system for grazing during 3 years. The mixture of perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) was established by undersowing in spring...... barley (Hordeum vulgare L.) and maintained subsequently for two production years. Dinitrogen fixation was determined using the N-15 isotope dilution techniques and two labelling procedures. Using either pre-labelling of the soil with immobilisation of the N-15 by addition of a carbon source before...

  8. Dinitrogen fixation in white clover grown in pure stand and mixture with ryegrass estimated by the immobilized 15N isotope dilution method

    DEFF Research Database (Denmark)

    Jørgensen, F.V.; Jensen, E.S.; Schjørring, J.K.

    1999-01-01

    Dinitrogen fixation in white clover (Trifolium repens L.) grown in pure stand and mixture with perennial ryegrass (Lolium perenne L.) was determined in the field using N-15 isotope dilution and harvest of the shoots. The apparent transfer of clover N to perennial ryegrass was simultaneously...... assessed. The soil was labelled either by immobilizing N-15 in organic matter prior to establishment of the sward or by using the conventional labelling procedure in which N-15 fertilizer is added after sward establishment. Immobilization of N-15 in the soil organic matter has not previously been used...

  9. Early interspecific interference in the wheat/faba bean (Triticum aestivum/ Vicia faba ssp. minor and rapeseed/squarrosum clover (Brassica napus var. oleifera/Trifolium squarrosum intercrops

    Directory of Open Access Journals (Sweden)

    Paolo Benincasa

    2012-04-01

    Full Text Available Most of research on intercrops evaluate performances and interference between species on the basis of final yield, while little knowledge is available on the interference in early stages and at the root level, at least for cultivated intercrops. In fact, in the few studies on this subject species are often combined minding at experimental needs (e.g. common substrate, temperature and water requirements, easy root separation more than at their actual use in the farm. The present work evaluates interspecific interference during early developmental stages for two intercrops of agricultural interest: soft wheat-faba bean and rapeseed-squarrosum clover. Improving this knowledge would help intercrop growth modelling and rational cultivation. The experiments were carried out on soft wheat (Triticum aestivum, faba bean (Vicia faba var. minor, rapeseed (Brassica napus var. oleifera and squarrosum clover (Trifolium squarrosum, germinated and grown until 32 days after sowing (DAS as one-species stands or as wheat/faba bean and rapeseed/squarrosum clover intercrops, with different densities and proportions for the two species in each couple. Germination was studied in controlled-temperature chamber, plantlet growth was studied on pots in the greenhouse. During germination no interspecific interference was observed for both wheat/faba bean and rapeseed/squarrosum clover intercrops. During plantlet growth, interspecific interference occurred in both intercrops causing variations in whole plant and root dry matter accumulation. In the wheat/faba bean intercrop each species suffered from the competitive effect of the companion species and faba bean was the dominant species when present in lower proportion than wheat. The unexpectedly high aggressivity of faba bean may be explained either with the greater seed size that could have represented an initial advantage within the short duration of the experiment or with the competition towards wheat for substrate N

  10. Glutathione S-transferase gene polymorphisms in presbycusis.

    Science.gov (United States)

    Ateş, Nurcan Aras; Unal, Murat; Tamer, Lülüfer; Derici, Ebru; Karakaş, Sevim; Ercan, Bahadir; Pata, Yavuz Selim; Akbaş, Yücel; Vayisoğlu, Yusuf; Camdeviren, Handan

    2005-05-01

    Glutathione and glutathione-related antioxidant enzymes are involved in the metabolism and detoxification of cytotoxic and carcinogenic compounds as well as reactive oxygen species. Reactive oxygen species generation occurs in prolonged relative hypoperfusion conditions such as in aging. The etiology of presbycusis is much less certain; however, a complex genetic cause is most likely. The effect of aging shows a wide interindividual range; we aimed to investigate whether profiles of (glutathione S-transferase (GST) M1, T1 and P1 genotypes may be associated with the risk of age-related hearing loss. We examined 68 adults with presbycusis and 69 healthy controls. DNA was extracted from whole blood, and the GSTM1, GSTT1 and GSTP1 polymorphisms were determined using a real-time polymerase chain reaction and fluorescence resonance energy transfer with a Light-Cycler Instrument. Associations between specific genotypes and the development of presbycusis were examined by use of logistic regression analyses to calculate odds ratios and 95% confidence intervals. Gene polymorphisms at GSTM1, GSTT1, and GSTP1 in subjects with presbycusis were not significantly different than in the controls (p > 0.05). Also, the combinations of different GSTM1, GSTT1, and GSTP1 genotypes were not an increased risk of presbycusis (p > 0.05). We could not demonstrate any significant association between the GSTM1, GSTT1, and GSTP1 polymorphism and age-related hearing loss in this population. This may be because of our sample size, and further studies need to investigate the exact role of GST gene polymorphisms in the etiopathogenesis of the presbycusis.

  11. Effect of maturity and conservation of grass/clover on digestibility and rumen pH in heifers

    DEFF Research Database (Denmark)

    Koch, A.S.; Nørgaard, P.; Weisbjerg, Martin Riis

    2013-01-01

    The study aimed to evaluate effects of maturity and conservation of primary growth grass/clover on apparent digestibility and rumen pH. Two batches of mixed ryegrass, red and white clover harvested in 2009 on May 9 and 25 were conserved as either silage or hay. The forages early silage (ES) and hay...... (EH), and late silage (LS) and hay (LH) had DM contents of 45, 84, 25 and 83%, and NDF contents of 32, 44, 42 and 50% of DM, respectively. Forages were fed as sole feed to four Jersey heifers of 435±30 kg BW in a 4×4 Latin square experiment. Feeding level was 90% of individual ad libitum intake......, divided in two daily meals at 0800 and 1530 h. Potentially digestible NDF (DNDF) was determined after 288 h in situ. Apparent digestibility of OM and NDF was estimated using Cr3O2 as marker. Rumen fluid pH in the medial and ventral rumen was measured with 1 h intervals from 0730 to 1530 h. Data...

  12. Effect of Mycorrhizal Fungi and Trifluralin Herbicide on Emergence, Growth and Root Colonization of Clover (Trifolium repens L.

    Directory of Open Access Journals (Sweden)

    Hassan Shahgholi

    2016-09-01

    Full Text Available Introduction: Herbicides, despite of their control of weeds, have the potential to affect sensitive crops in rotation and also beneficial non-targeted soil microbes including vesicular arbuscular mycorrhiza (VAM fungi (6. AM fungi can increase the growth of crops through increasing uptake of phosphorus and insoluble micronutrients, and indirectly by improving soil quality parameters (30. However, several authors have reported different effects of herbicides on VAM symbiosis, which ranges from no adverse effects to slightly or highly toxic effects (6. Pesticides have also been reported to stimulate colonization of plant roots by AM fungi (27. Therefore, the objective of this study was to investigate the interaction effects of mycorrhizal fungi and Trifluralin herbicide on the growth and root colonization of clover. Materials and Methods: A factorial experiment was arranged in randomized complete block design with three replicates at the College of Agricultural, University of Shahrood during 2012. Treatments were included three levels of mycorrhiza inoculation, M1: non mycorrhiza (control, M2: Glommus mosseae and M3: Glommus intraradices and herbicide treatments were included four levels of Trifluralin(T1: 0, T2: 1000, T3: 1500 and T4: 2000 ml ha-1. In mycorrhizal treatments, 20 g inoculums were thoroughly mixed with soil. Seeds of clover (Trifolium repens L. were sown in the pots maintained near the field in order to provide normal environmental conditions. Seedlings were thinned to two plants per pot at three leaf stages. At the time of harvesting, the emergence and growth characteristics of clover and root colonization was also registered. Statistical analyses of data were performed with statistical software MSTATC. Significant differences between means refer to the probability level of 0.05 calculated by LSD test. Results and Discussion: The results showed that emergence, uniformity (EU values decreased and time to 10% (D10 and 90% (D90 of

  13. The Growth Rate and Efficiency of Rumen Microbial Protein Digestion of Red Clover Silage (Trifolium pratense cv. Sabatron)

    International Nuclear Information System (INIS)

    Asih Kurniawati

    2004-01-01

    (Trifolium pratense cv. Sabatron). Red clover silage supplemented with different level of carbohydrates has been examined using the in-vitro gas production technique. Cumulative gas production, hydro.gen sulfite production, and ammonia was followed and used as indicators of microbial growth rate and extent of protein degradation. Microbial nitrogen production, VFA, and efficiency microbial production was used as indicator of nitrogen use efficiency. 15 N was used as a microbial marker to estimate the amount of nitrogen incorporation into microbial protein. Supplementation of Red clover with increasing 5 levels; 0 g; 0.625 g; 0.15 g; 0.225 g and 0.3 g of maize starch led to graded increase in microbial growth and protein degradation. This was reflected in the increasing gas production and the accumulation of hydrogen sulfite. Diurnal change in ammonia production reflected the microbial utilization of ammonia for protein synthesis. Protein microbe (P<0.001) as VFA (P<0.001) increased due to carbohydrate addition as well as utilization of nitrogen (P<0.001). There was also the efficiency of nitrogen utilization which increased significantly. This result suggested that energy supply can increased efficiency of nitrogen use in the rumen and may reduce nitrogen losses into the environment. (author)

  14. Response of biomass and nitrogen yield of white clover to radiation and atmospheric CO2 concentration

    International Nuclear Information System (INIS)

    Manderscheid, R.; Bender, J.; Schenk, U.; Weigel, H.J.

    1997-01-01

    The objectives of the present study were to test (i) whether the effect of season-long CO 2 enrichment on plant dry matter production of white clover (Trifolium repens cv. Karina) depends on the temperature or can solely be explained by changes in radiation use efficiency, and (ii) whether the atmospheric CO 2 concentration affects the relationship between tissue %N and plant biomass. Plants were grown in pots with adequate nutrient and water supply and were exposed to ambient and above ambient CO 2 concentrations (approximately +80 ppm, +160 ppm, +280 ppm) in open-top chambers for two seasons. Nitrogen fertilizer was given only before the experiment started to promote N 2 fixation. Plants were clipped to a height of 5 cm, when the canopy had reached a height of about 20 cm and when the CO 2 effect had not been diminished due to self-shading of the leaves. Photon exposure (400–700 nm) measured above the canopy was linearly related to the above ground biomass, the leaf area index and the nitrogen yield (r 2 > 0.94). The slopes of the curves depended on the CO 2 concentration. Since most of the radiation (>90%) was absorbed by the foliage, the slopes were used to calculate the CO 2 effect on the radiation use efficiency of biomass production, which is shown to increase curvilinearly between 380 and 660 ppm CO 2 from 2.7 g MJ −1 to 3.9 g MJ −1 . CO 2 enrichment increased above ground biomass by increasing the leaf number, the individual leaf weight and the leaf area; specific leaf weight was not affected. The relative CO 2 response varied between harvests; there was a slight but not significant positive relationship with mean daytime temperature. At the beginning of the season, plant nitrogen concentration in the above ground biomass was decreased by CO 2 enrichment. However, at later growth stages, when the plants depended solely on N 2 fixation, nitrogen concentration was found to be increased when the nitrogen concentration value was adjusted for the decrease

  15. Human glutathione transferases catalyzing the bioactivation of anticancer thiopurine prodrugs.

    Science.gov (United States)

    Eklund, Birgitta I; Gunnarsdottir, Sjofn; Elfarra, Adnan A; Mannervik, Bengt

    2007-06-01

    cis-6-(2-Acetylvinylthio)purine (cAVTP) and trans-6-(2-acetylvinylthio)guanine (tAVTG) are thiopurine prodrugs provisionally inactivated by an alpha,beta-unsaturated substituent on the sulfur of the parental thiopurines 6-mercaptopurine (6-MP) and 6-thioguanine (6-TG). The active thiopurines are liberated intracellularly by glutathione (GSH) in reactions catalyzed by glutathione transferases (GSTs) (EC 2.5.1.18). Catalytic activities of 13 human GSTs representing seven distinct classes of soluble GSTs have been determined. The bioactivation of cAVTP and tAVTG occurs via a transient addition of GSH to the activated double bond of the S-substituent of the prodrug, followed by elimination of the thiopurine. The first of these consecutive reactions is rate-limiting for thiopurine release, but GST-activation of this first addition is shifting the rate limitation to the subsequent elimination. Highly active GSTs reveal the transient intermediate, which is detectable by UV spectroscopy and HPLC analysis. LC/MS analysis of the reaction products demonstrates that the primary GSH conjugate, 4-glutathionylbuten-2-one, can react with a second GSH molecule to form the 4-(bis-glutathionyl)butan-2-one. GST M1-1 and GST A4-4 were the most efficient enzymes with tAVTG, and GST M1-1 and GST M2-2 had highest activity with cAVTP. The highly efficient GST M1-1 is polymorphic and is absent in approximately half of the human population. GST P1-1, which is overexpressed in many cancer cells, had no detectable activity with cAVTP and only minor activity with tAVTG. Other GST-activated prodrugs have targeted GST P1-1-expressing cancer cells. Tumors expressing high levels of GST M1-1 or GST A4-4 can be predicted to be particularly vulnerable to chemotherapy with cAVTP or tAVTG.

  16. Breeding biologies, pollinators and seed beetles of two prairie-clovers, Dalea ornata and D. searlsiae (Fabaceae: Amorpheae), from the Intermountain West USA

    Science.gov (United States)

    Two prairie-clovers, Dalea ornata and D. searlsiae, are perennial forbs that flower during early summer throughout the Colombia Plateau and Great Basin of the western USA, respectively. Their seed is desirable for use in rangeland restoration. We experimentally characterized the breeding biologies ...

  17. Effects of forest management on running buffalo clover (Trifolium stoloniferum Muhl. Ex A. Eaton) distribution and abundance in the Fernow Experimental Forest

    Science.gov (United States)

    J.Q. Burkhart; J.R. Rentch; T.M. Schuler

    2013-01-01

    Identifying habitat preferences of species of concern is fundamental to the practice of conservation, but disturbances and other environmental processes can substantially affect suitability. Trifolium stoloniferum, or running buffalo clover, is a federally endangered plant species that occurs on the Fernow Experimental Forest in West Virginia....

  18. The combined effect of fertiliser nitrogen and phosphorus on herbage yield and change in soil nutrients of a grass/clover and grass-only sward

    NARCIS (Netherlands)

    Schils, R.L.M.; Snijders, P.J.M.

    2004-01-01

    The combined effect of reduced nitrogen ( N ) and phosphorus ( P ) application on the production of grass- only and grass/ clover swards was studied in a five- year cutting experiment on a marine clay soil, established on newly sown swards. Furthermore, changes in soil N, P and carbon ( C ) were

  19. Disconnected quark loop contributions to nucleon observables using Nf=2 twisted clover fermions at the physical value of the light quark mass

    International Nuclear Information System (INIS)

    Abdel-Rehim, Abdou; Kallidonis, Christos; Koutsou, Giannis

    2015-11-01

    We compute the disconnected quark loops contributions entering the determination of nucleon observables, by using a N f =2 ensemble of twisted mass fermions with a clover term at a pion mass m π =133 MeV. We employ exact deflation and implement all calculations in GPUs, enabling us to achieve large statistics and a good signal.

  20. Disconnected quark loop contributions to nucleon observables using N{sub f}=2 twisted clover fermions at the physical value of the light quark mass

    Energy Technology Data Exchange (ETDEWEB)

    Abdel-Rehim, Abdou; Kallidonis, Christos; Koutsou, Giannis [Cyprus Institute, Nicosia (Cyprus). Computation-based Science and Technology Research Center; Alexandrou, Constantia; Constantinou, Martha; Hadjiyiannakou, Kyriakos [Cyprus Institute, Nicosia (Cyprus). Computation-based Science and Technology Research Center; Cyprus Univ. (Cyprus). Dept. of Physics; Jansen, Karl [DESY Zeuthen (Germany). NIC; Aviles-Casco, Alejandro Vaquero [INFN Sezione di Milano-Bicocca, Milano (Italy)

    2015-11-15

    We compute the disconnected quark loops contributions entering the determination of nucleon observables, by using a N{sub f}=2 ensemble of twisted mass fermions with a clover term at a pion mass m{sub π}=133 MeV. We employ exact deflation and implement all calculations in GPUs, enabling us to achieve large statistics and a good signal.

  1. Amino acid profile of metabolisable protein in lactating dairy cows is affected by dry matter concentration in grass-clover silage

    DEFF Research Database (Denmark)

    Johansen, Marianne; Lund, Peter; Weisbjerg, Martin Riis

    2018-01-01

    Our previous study showed that supply of metabolisable protein (MP) to lactating dairy cows increased with increasing dry matter (DM) concentration in grass-clover silage. The aim of this study was to examine how amino acid (AA) profile of MP was affected by silage DM concentration. Eight grass-c...

  2. Isolation of Burkholderia cepacia JB12 from lead- and cadmium-contaminated soil and its potential in promoting phytoremediation with tall fescue and red clover.

    Science.gov (United States)

    Jin, Zhong Min; Sha, Wei; Zhang, Yan Fu; Zhao, Jing; Ji, Hongyang

    2013-07-01

    Phytoremediation combined with suitable microorganisms and biodegradable chelating agents can be a means of reclaiming lands contaminated by toxic heavy metals. We investigated the ability of a lead- and cadmium-resistant bacterial strain (JB12) and the biodegradable chelator ethylenediamine-N,N'-disuccinic acid (EDDS) to improve absorption of these metals from soil by tall fescue and red clover. Strain JB12 was isolated from contaminated soil samples, analysed for lead and cadmium resistance, and identified as Burkholderia cepacia. Tall fescue and red clover were grown in pots to which we added JB12, (S,S)-EDDS, combined JB12 and EDDS, or water only. Compared with untreated plants, the biomass of plants treated with JB12 was significantly increased. Concentrations of lead and cadmium in JB12-treated plants increased significantly, with few exceptions. Plants treated with EDDS responded variably, but in those treated with combined EDDS and JB12, heavy metal concentrations increased significantly in tall fescue and in the aboveground parts of red clover. We conclude that JB12 is resistant to lead and cadmium. Its application to the soil improved the net uptake of these heavy metals by experimental plants. The potential for viable phytoremediation of lead- and cadmium-polluted soils with tall fescue and red clover combined with JB12 was further enhanced by the addition of EDDS.

  3. Annual maize and perennial grass-clover strip cropping for increased resource use efficiency and productivity using organic farming practice as a model

    DEFF Research Database (Denmark)

    Hauggaard-Nielsen, Henrik; Johansen, Anders; Carter, Mette Sustmann

    2013-01-01

    A cropping system was designed to fulfill the increasing demand for biomass for food and energy without decreasing long term soil fertility. A field experiment was carried out including alternating strips of annual maize (Zea mays L.) and perennial ryegrass (Lolium perenne L.) – clover (Trifolium...

  4. Short-term effects of a dung pat on N2 fixation and total N uptake in a perennial ryegrass/white clover mixture

    DEFF Research Database (Denmark)

    Jørgensen, F.V.; Jensen, E.S.

    1997-01-01

    The short-term effects of a simulated cattle dung pat on N-2 fixation and total uptake of N in a perennial ryegrass/white clover mixture was studied in a container experiment using sheep faeces mixed with water to a DM content of 13%. We used a new N-15 cross-labelling technique to determine...

  5. Biochanin A (an isoflavone produced by red clover) promotes weight gain of steers grazed in mixed grass pastures and fed dried-distillers grains

    Science.gov (United States)

    Biochanin A (BCA) is an isoflavone produced by red clover (Trifloium pratense L.) that can inhibit hyper-ammonia producing bacteria (HAB) to reduce deamination in the rumen and increase the feed amino acids available for gastric digestion. An in vitro experiment was conducted to evaluate the effect...

  6. The effect of additives in silages of pure timothy and timothy mixed with red clover on chemical composition and in vitro rumen fermentation characteristics

    NARCIS (Netherlands)

    Hetta, M.; Cone, J.W.; Gustavsson, A.M.; Martinsson, K.

    2003-01-01

    The aim was to compare the effects of additives on direct cut silages of pure timothy and timothy mixed with tetraploid red clover. First and second growth cuts were ensiled during three consecutive years, 1994, 1995 and 1996, either without any additive or with the addition of formic acid, or

  7. Transcription of Biotic Stress Associated Genes in White Clover (Trifolium repens L.) Differs in Response to Cyst and Root-Knot Nematode Infection.

    Science.gov (United States)

    Islam, Afsana; Mercer, Chris F; Leung, Susanna; Dijkwel, Paul P; McManus, Michael T

    2015-01-01

    The transcription of four members of the Kunitz proteinase inhibitor (KPI) gene family of white clover (Trifolium repens L.), designated as Tr-KPI1, Tr-KPI2, Tr-KPI4 and Tr-KPI5, was investigated at both local infection (roots) and systemic (leaf tissue) sites in white clover in response to infection with the clover root knot nematode (CRKN) Meloidogyne trifoliophila and the clover cyst nematode (CCN) Heterodera trifolii. Invasion by the CRKN resulted in a significant decrease in transcript abundance of Tr-KPI4 locally at both 4 days post-infection (dpi) and at 8 dpi, and an increase in transcription of Tr-KPI1 systemically at 8 dpi. In contrast, an increase in transcript abundance of all four Tr-KPI genes locally at 4 and 8 dpi, and an increase of Tr-KPI1, Tr-KPI2, and Tr-KPI5 at 8 dpi systemically was observed in response to infection with the CCN. Challenge of a resistant (R) genotype and a susceptible (S) genotype of white clover with the CCN revealed a significant increase in transcript abundance of all four Tr-KPI genes locally in the R genotype, while an increase in abundance of only Tr-KPI1, Tr-KPI2, and Tr-KPI5 was observed in the S genotype, and only at 4 dpi. The transcript abundance of a member of the1-AMINOCYCLOPROPANE-1-CARBOXYLATE (ACC) SYNTHASE gene family from white clover (Tr-ACS1) was significantly down-regulated locally in response to CRKN infection at 4 and 8 dpi and at 4 dpi, systemically, while abundance increased locally and systemically at 8 dpi in response to CCN challenge. Conversely, the abundance of the jasmonic acid (JA) signalling gene, CORONATINE-INSENSITIVE PROTEIN 1 from white clover (Tr-COI1) increased significantly at 8 dpi locally in response to CRKN infection, but decreased at 8 dpi in response to CCN infection. The significance of this differential regulation of transcription is discussed with respect to differences in infection strategy of the two nematode species.

  8. Microbial protein synthesis, digestion and lactation responses of cows to grass or grass-red clover silage diet supplemented with barley or oats

    Directory of Open Access Journals (Sweden)

    A. VANHATALO

    2008-12-01

    Full Text Available The study was conducted to evaluate effects of silage type (grass-red clover vs. pure grass and grain supplement (oats vs. barley on rumen fermentation, post-ruminal nutrient flows, diet digestion and milk production. Four primiparous Finnish Ayrshire cows fitted with cannulae in the rumen and duodenum were used in a 4 × 4 Latin square experiment with four 28-d experimental periods and 2 × 2 factorial arrangements of treatments. Using red clover-containing (40% silage rather than pure grass silage had minor effects on rumen fermentation or diet digestion but increased non-ammonia nitrogen (N flow in terms of increased flows of microbial and dietary N entering to the small intestine. This was reflected as a reduced ruminal N degradability on grass-red clover diets. Furthermore, grass-red clover diets in comparison to grass silage diets increased milk lactose concentration and yields of milk, protein and lactose. Feeding oats in replacement for barley had minor effects on rumen fermentation or post-ruminal non-ammonia N flows but reduced digestibility of organic matter and neutral detergent fibre in the diet. Using oats rather than barley increased yields of milk and lactose but reduced milk protein concentration. Oats also increased proportions of C18:0 and C18:1 in milk fat and reduced those of C10:0 to C16:0. It is concluded that inclusion of red clover and replacement of barley with oats in grass silage based diets have beneficial effects in dairy cow production.;

  9. Utilization of protein in red clover and alfalfa silages by lactating dairy cows and growing lambs.

    Science.gov (United States)

    Broderick, Glen A

    2018-02-01

    Feeding trials were conducted with lactating cows and growing lambs to quantify effects of replacing dietary alfalfa silage (AS) with red clover silage (RCS) on nutrient utilization. The lactation trial had a 2 × 4 arrangement of treatments: AS or RCS fed with no supplement, rumen-protected Met (RPM), rumen-protected Lys (RPL), or RPM plus RPL. Grass silage was fed at 13% of dry matter (DM) with AS to equalize dietary neutral detergent fiber (NDF) and crude protein contents. All diets contained (DM basis) 5% corn silage and 16% crude protein. Thirty-two multiparous (4 ruminally cannulated) plus 16 primiparous Holstein cows were blocked by parity and days in milk and fed diets as total mixed rations in an incomplete 8 × 8 Latin square trial with four 28-d periods. Production data (over the last 14 d of each period) and digestibility and excretion data (at the end of each period) were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Although DM intake was 1.2 kg/d greater on AS than RCS, milk yield and body weight gain were not different. However, yields of fat and energy-corrected milk as well as milk content of fat, true protein, and solids-not-fat were greater on AS. Relative to AS, feeding RCS increased milk and energy-corrected milk yield per unit of DM intake, milk lactose content, and apparent N efficiency and reduced milk urea. Relative to AS, apparent digestibility of DM, organic matter, NDF, and acid detergent fiber were greater on RCS, whereas apparent and estimated true N digestibility were lower. Urinary N excretion and ruminal concentrations of ammonia, total AA, and branched-chain volatile fatty acids were reduced on RCS, indicating reduced ruminal protein degradation. Supplementation of RPM increased intake, milk true protein, and solids-not-fat content and tended to increase milk fat content. There were no silage × RPM interactions, suggesting that RPM was equally limiting on both AS and RCS. Supplementation of RPL did not

  10. Presence of a Ca2+-sensitive CDPdiglyceride-inositol transferase in canine cardiac sarcoplasmic reticulum

    International Nuclear Information System (INIS)

    Kasinathan, C.; Kirchberger, M.A.

    1988-01-01

    Sarcoplasmic reticulum (SR) and plasma membranes from canine left ventricle were used to evaluate the presence of the enzyme CDPdiglyceride-inositol transferase in these membranes. (K + ,-Ca 2+ )-ATPase activity, a marker for SR, was 79.2 +/- 5.0 (SE) and 11.2 +/- 2.0 μmol x mg -1 x h -1 in SR and plasma membrane preparations, respectively, and (Na + , K + )-ATPase activity, a marker for plasma membranes, was 5.6 +/- 1.2 and 99.2 +/- 8.0 μmol x mg -1 x h -1 , respectively. Contamination of SR and plasma membrane preparations by mitochondria was estimated to be 2% and 8%, respectively, and by Golgi membranes, 0.9% and 1.8%, respectively. The transferase activity detected in the plasma membrane preparation could be accounted for largely, but not entirely, by contaminating SR membranes. The pH optimum for the SR transferase activity was between 8.0 and 9.0. Ca 2+ inhibited the enzyme, half-maximal inhibition occurring at about 10 μM Ca 2+ . No loss of [ 3 H]PtdIns could be detected when membranes were incubated in the presence or absence of Ca 2+ . The Ca 2+ inhibition of the transferase was noncompetitive with respect to CDP-dipalmitin while that with respect to myo-inositol was slightly noncompetitive at low [Ca 2+ ] and became uncompetitive at higher [Ca 2+ ]. It is concluded that CDPdiglyceride-inositol transferase is present on SR membranes and is sensitive to micromolar Ca 2+ . The data are consistent with a putative role for the inhibition of the SR transferase by Ca 2+ and acidic pH in the protection of the SR against calcium overload in ischemic myocardium

  11. Evaluation of physical structure value in spring-harvested grass/clover silage and hay fed to heifers

    DEFF Research Database (Denmark)

    Schulze, A.K.S.; Nørgaard, P.; Byskov, M.V.

    2015-01-01

    The physical structure value of conserved grass/clover forages of spring harvest was evaluated by assessing effects of harvest time, conservation method, iNDF/NDF ratio and NDF intake (NDFI) per kg BW on chewing activity and fecal particle size in dairy heifers. A mixed sward consisting of ryegrass...... of 315, 436, 414 and 503 g/kg DM, respectively. Forages were fed as sole feed to four Jersey heifers of 435±30 kg BW in a 4×4 Latin square experiment. Feeding level was 90% of individual ad libitum intake, divided equally across two daily meals offered at 0800 and 1530 h. Chewing activity was estimated...... from recorded jaw movements (JM) oscillations continuously logged for 96 h and summarized per 24 h as mean effective rumination time and eating time. Eating behavior was further observed during four 20-min test meals. Weight proportion of large feces particles (>1.0 mm) and geometric mean fecal...

  12. Effects of grazing strategy on limiting nitrate leaching in grazed grass-clover pastures on coarse sandy soil

    DEFF Research Database (Denmark)

    Hansen, Elly Møller; Eriksen, Jørgen; Søegaard, Karen

    2012-01-01

    -term mean. The experiment was initiated in a 4-yr-old grass-clover sward in south Denmark. Three treatments were as follows grazing only (G), spring cut followed by grazing (CG) and both spring and autumn cuts with summer grazing (CGC). Nitrate leaching was calculated by extracting water isolates from 80 cm......Urinations of ruminants on grazed pastures increase the risk of nitrate leaching. The study investigated the effect of reducing the length of the grazing season on nitrate leaching from a coarse sandy, irrigated soil during 2006–2007 and 2007–2008. In both years, precipitation was above the long...... depth using ceramic suction cups. Because of considerable variation in measured nitrate concentrations, the 32 installed suction cups per treatment were insufficient to reveal differences between treatments. However, weighted nitrate leaching estimations for G, CG and CGC showed estimated mean nitrate N...

  13. Does introduction of clover in an agricultural grassland affect the food base and functional diversity of Collembola?

    DEFF Research Database (Denmark)

    D'Annibale, Alessandra; Sechi, Valentina; Larsen, Thomas

    2017-01-01

    plots with either perennial ryegrass (Lolium perenne L.), whiteclover (Trifolium repens L.) or a mixture of both in a Danish agricultural grassland 6 and 14 months after establishing the leys (September and May, respectively). Diet preferences were investigated via stable isotope analyses (SIA...... in the white clover than ryegrass plots. Changes in taxa specific density and traits distribution as a response to the C:N ratio of plant material, suggest that plant material quality was the main factor affecting the collembolan community,especially when comparing the two sampling occasions. Functional...... richness decreased under conditions of low quality material. In contrast to our hypothesis, population densities did not increase under mixture treatment and functional richness decreased. Our results suggest that habitat changes, via different plant composition, can affect some functional groups, having...

  14. Epidermal growth factor regulation of glutathione S-transferase gene expression in the rat is mediated by class Pi glutathione S-transferase enhancer I.

    OpenAIRE

    Matsumoto, M; Imagawa, M; Aoki, Y

    2000-01-01

    Using chloramphenicol acetyltransferase assays we showed that epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha), and 3,3',4,4',5-pentachlorobiphenyl (PenCB) induce class Pi glutathione S-transferase (GSTP1) in primary cultured rat liver parenchymal cells. GSTP1 enhancer I (GPEI), which is required for the stimulation of GSTP1 expression by PenCB, also mediates EGF and TGF alpha stimulation of GSTP1 gene expression. However, hepatocyte growth factor and insulin did no...

  15. Rhizobium leguminosarum bv. trifolii rosR is required for interaction with clover, biofilm formation and adaptation to the environment

    Directory of Open Access Journals (Sweden)

    Piersiak Tomasz

    2010-11-01

    Full Text Available Abstract Background Rhizobium leguminosarum bv. trifolii is a symbiotic nitrogen-fixing bacterium that elicits nodules on roots of host plants Trifolium spp. Bacterial surface polysaccharides are crucial for establishment of a successful symbiosis with legumes that form indeterminate-type nodules, such as Trifolium, Pisum, Vicia, and Medicago spp. and aid the bacterium in withstanding osmotic and other environmental stresses. Recently, the R. leguminosarum bv. trifolii RosR regulatory protein which controls exopolysaccharide production has been identified and characterized. Results In this work, we extend our earlier studies to the characterization of rosR mutants which exhibit pleiotropic phenotypes. The mutants produce three times less exopolysaccharide than the wild type, and the low-molecular-weight fraction in that polymer is greatly reduced. Mutation in rosR also results in quantitative alterations in the polysaccharide constituent of lipopolysaccharide. The rosR mutants are more sensitive to surface-active detergents, antibiotics of the beta-lactam group and some osmolytes, indicating changes in the bacterial membranes. In addition, the rosR mutants exhibit significant decrease in motility and form a biofilm on plastic surfaces, which differs significantly in depth, architecture, and bacterial viability from that of the wild type. The most striking effect of rosR mutation is the considerably decreased attachment and colonization of root hairs, indicating that the mutation affects the first stage of the invasion process. Infection threads initiate at a drastically reduced rate and frequently abort before they reach the base of root hairs. Although these mutants form nodules on clover, they are unable to fix nitrogen and are outcompeted by the wild type in mixed inoculations, demonstrating that functional rosR is important for competitive nodulation. Conclusions This report demonstrates the significant role RosR regulatory protein plays in

  16. Purification of human hepatic glutathione S-transferases and the development of a radioimmunoassay for their measurement in plasma

    International Nuclear Information System (INIS)

    Hayes, J.D.; Gilligan, D.; Beckett, G.J.

    1983-01-01

    A purification scheme is described for six human hepatic glutathione S-transferases from a single liver. Five of the transferases comprised Ya monomers and had a molecular mass of 44000. The remaining enzyme comprised Yb monomers and had a molecular mass of 47000. Data are presented demonstrating that there are at least two distinct Ya monomers. A radioimmunoassay has been developed that has sufficient precision and sensitivity to allow direct measurement of glutathione S-transferase concentrations in unextracted plasma. A comparison of aminotransferase and glutathione S-transferase levels, in three patients who had taken a paracetamol overdose, indicated that glutathione S-transferase measurements provided a far more sensitive index of hepatocellular integrity than the more conventional aminotransferase measurements. (Auth.)

  17. Purification of human hepatic glutathione S-transferases and the development of a radioimmunoassay for their measurement in plasma

    Energy Technology Data Exchange (ETDEWEB)

    Hayes, J.D.; Gilligan, D.; Beckett, G.J. (Edinburgh Univ. (UK). Dept. of Clinical Chemistry); Chapman, B.J. (Royal Infirmary, Edinburgh (UK))

    1983-10-31

    A purification scheme is described for six human hepatic glutathione S-transferases from a single liver. Five of the transferases comprised Ya monomers and had a molecular mass of 44000. The remaining enzyme comprised Yb monomers and had a molecular mass of 47000. Data are presented demonstrating that there are at least two distinct Ya monomers. A radioimmunoassay has been developed that has sufficient precision and sensitivity to allow direct measurement of glutathione S-transferase concentrations in unextracted plasma. A comparison of aminotransferase and glutathione S-transferase levels, in three patients who had taken a paracetamol overdose, indicated that glutathione S-transferase measurements provided a far more sensitive index of hepatocellular integrity than the more conventional aminotransferase measurements.

  18. Glutathione s-transferase isoenzymes in relation to their role in detoxification of xenobiotics

    NARCIS (Netherlands)

    Vos, R.M.E.

    1989-01-01

    The glutathione S-transferases (GST) are a family of isoenzymes serving a major part in the biotransformation of many reactive compounds. The isoenzymes from rat, man and mouse are divided into three classes, alpha, mu and pi, on the basis of similar structural and enzymatic

  19. Isolation and characterization of an auxin-inducible glutathione S-transferase gene of Arabidopsis thaliana

    NARCIS (Netherlands)

    Kop, D.A.M. van der; Schuyer, M.; Scheres, B.J.G.; Zaal, B.J. van der; Hooykaas, P.J.J.

    1996-01-01

    Genes homologous to the auxin-inducible Nt103 glutathione S-transferase (GST) gene of tobacco, were isolated from a genomic library of Arabidopsis thaliana. We isolated a λ clone containing an auxin-inducible gene, At103-1a, and part of a constitutively expressed gene, At103-1b. The coding regions

  20. Global deletion of glutathione S-Transferase A4 exacerbates developmental nonalcoholic steatohepatitis

    Science.gov (United States)

    We established a mouse model of developmental nonalcoholic steatohepatitis (NASH) by feeding a high polyunsaturated fat liquid diet to female glutathione-S-transferase 4-4 (Gsta4-/-)/peroxisome proliferator activated receptor a (Ppara-/-) double knockout 129/SvJ mice for 12 weeks from weaning. We us...

  1. Involvement of human glutathione S-transferase isoenzymes in the conjugation of cyclophosphamide metabolites with glutathione

    NARCIS (Netherlands)

    Dirven, H.A.A.M.; Ommen, B. van; Bladeren, P.J. van

    1994-01-01

    Alkylating agents can be detoxified by conjugation with glutathione (GSH). One of the physiological significances of this lies in the observation that cancer cells resistant to the cytotoxic effects of alkylating agents have higher levels of GSH and high glutathione S-transferase (GST) activity.

  2. The role of glutathione S-transferase and claudin-1 gene polymorphisms in contact sensitization

    DEFF Research Database (Denmark)

    Ross-Hansen, K; Linneberg, A; Johansen, J D

    2013-01-01

    BACKGROUND: Contact sensitization is frequent in the general population and arises from excessive or repeated skin exposure to chemicals and metals. However, little is known about its genetic susceptibility. OBJECTIVES: To determine the role of polymorphisms of glutathione S-transferase (GST) genes...

  3. Effects of curcumin on cytochrome P450 and glutathione S-transferase activities in rat liver.

    NARCIS (Netherlands)

    Oetari, S.; Sudibyo, M.; Commandeur, J.N.M.; Samhoedi, R.; Vermeulen, N.P.E.

    1996-01-01

    The stability of curcumin, as well as the interactions between curcumin and cytochrome P450s (P450s) and glutathione S-transferases (GSTs) in rat liver, were studied. Curcumin is relatively unstable in phosphate buffer at pH 7.4. The stability of curcumin was strongly improved by lowering the pH or

  4. Bisubstrate Kinetics of Glutathione S-Transferase: A Colorimetric Experiment for the Introductory Biochemistry Laboratory

    Science.gov (United States)

    Stefanidis, Lazaros; Scinto, Krystal V.; Strada, Monica I.; Alper, Benjamin J.

    2018-01-01

    Most biochemical transformations involve more than one substrate. Bisubstrate enzymes catalyze multiple chemical reactions in living systems and include members of the transferase, oxidoreductase, and ligase enzyme classes. Working knowledge of bisubstrate enzyme kinetic models is thus of clear importance to the practicing biochemist. However,…

  5. Association study on glutathione S-transferase omega 1 and 2 and familial ALS

    NARCIS (Netherlands)

    van de Giessen, Elsmarieke; Fogh, Isabella; Gopinath, Sumana; Smith, Bradley; Hu, Xun; Powell, John; Andersen, Peter; Nicholson, Garth; Al Chalabi, Ammar; Shaw, Christopher E.

    2008-01-01

    Glutathione S-transferase omega 1 and 2 (GSTO1 and 2) protect from oxidative stress, a possible pathogenic mechanism underlying the pathogenesis of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. Significant association of age of onset in Alzheimer's

  6. Primary overproduction of urate caused by a partial deficiency of hypoxanthine-guanine phosphoribosyl transferase

    International Nuclear Information System (INIS)

    Cassidy, M.; Gregory, M.C.; Harley, E.H.

    1980-01-01

    Inherited enzyme deficiencies are found in a small proportion of patients with gout who produce an excess of uric acid. The clinical, biochemical and therapeutic aspects of a case of hyperuricaemia caused by an atypical mutant hypoxanthine-guanine phophoribosyl transferase are presented. Urate overproduction was moderate and controlled by allopurinol therapy

  7. Glutathione S-transferase genotype and p53 mutations in adenocarcinoma of the small intestine

    DEFF Research Database (Denmark)

    Pedersen, Lisbeth Nørum; Kaerlev, L; Stubbe Teglbjaerg, P

    2003-01-01

    Adenocarcinoma of the small intestine (ASI) is a rare disease of unknown aetiology. The glutathione S-transferase M1 (GSTM1) enzyme catalyses the detoxification of compounds involved in carcinogenesis of adenocarcinoma of the stomach, colon and lung, including constituents of tobacco smoke. We in...

  8. Antibiotic inhibition of the movement of tRNA substrates through a peptidyl transferase cavity

    DEFF Research Database (Denmark)

    Porse, B T; Rodriguez-Fonseca, C; Leviev, I

    1996-01-01

    The present review attempts to deal with movement of tRNA substrates through the peptidyl transferase centre on the large ribosomal subunit and to explain how this movement is interrupted by antibiotics. It builds on the concept of hybrid tRNA states forming on ribosomes and on the observed movem...

  9. Inhibition of rat, mouse, and human glutathione S-transferase by eugenol and its oxidation products

    NARCIS (Netherlands)

    Rompelberg, C.J.M.; Ploemen, J.H.T.M.; Jespersen, S.; Greef, J. van der; Verhagen, H.; Bladeren, P.J. van

    1996-01-01

    The irreversible and reversible inhibition of glutathione S-transferases (GSTs) by eugenol was studied in rat, mouse and man. Using liver cytosol of human, rat and mouse, species differences were found in the rate of irreversible inhibition of GSTs by eugenol in the presence of the enzyme

  10. Functional characterization of glutathione S-transferases associated with insecticide resistance in Tetranychus urticae

    NARCIS (Netherlands)

    Pavlidi, N.; Tseliou, V.; Riga, M.; Nauen, R.; Van Leeuwen, T.; Labrou, N.E.; Vontas, J.

    2015-01-01

    The two-spotted spider mite Tetranychus urticae is one of the most important agricultural pests world-wide. It is extremely polyphagous and develops resistance to acaricides. The overexpression of several glutathione S-transferases (GSTs) has been associated with insecticide resistance. Here, we

  11. Microbial biomass, microbial diversity, soil carbon storage, and stability after incubation of soil from grass-clover pastures of different age

    DEFF Research Database (Denmark)

    Müller-Stöver, Dorette Sophie; Hauggaard-Nielsen, Henrik; Eriksen, Jørgen

    2012-01-01

    A laboratory incubation study with clover grass pasture soils of seven different ages (0, 1, 2, 3, 4, 5, and 16 production years) was carried out to determine initial soil carbon (C) and nitrogen (N) stocks and potentials for greenhouse gas emissions (N2O and CO2). Compared with the soil from...... the recently established pasture, an increase of total soil C and N was observed along with pasture age. Greenhouse gas emissions were low and not significantly different among the soils from younger pastures (0-5 years), but especially N2O emissions increased markedly in the soil from 16-year-old grass......-clover. Low emissions might mainly be due to an early C limitation occurring in the soils from younger pastures, which was also corroborated by decreasing levels of cold water-extractable C and early shifts within the microbial community. However, higher emissions from the old pasture soil were offset by its...

  12. Bicarbonate as tracer for plant assimilated C and homogeneity of 14C and 15N distribution in ryegrass and white clover tissue by alternative labeling approaches

    DEFF Research Database (Denmark)

    Rasmussen, Jim; Kusliene, Gedrime; Jacobsen, Ole Stig

    2013-01-01

    that 15N also had a heterogeneous distribution (up to two orders of magnitude). Conclusion Bicarbonate can efficiently be used to introduce 14C or 13C into plant via the leaf-labeling method. Both 14C and 15N showed heterogeneous distribution in the plant, although the distribution of 15N was more even......Aims: Application of carbon (C) and nitrogen (N) isotopes is an essential tool to study C and N flows in plant-soil-microorganisms systems. When targeting single plants in a community the tracers need to be added via e.g., leaf-labeling or stem-feeding approaches. In this study we: (i) investigated...... if bicarbonate can be used to introduce 14C (or 13C) into white clover and ryegrass, and (ii) compared the patterns of 14C and 15N allocation in white clover and ryegrass to evaluate the homogeneity of tracer distribution after two alternative labeling approaches. Methods Perennial ryegrass and white clover were...

  13. Arbuscular mycorrhiza enhanced arsenic resistance of both white clover (Trifolium repens Linn.) and ryegrass (Lolium perenne L.) plants in an arsenic-contaminated soil

    International Nuclear Information System (INIS)

    Dong Yan; Zhu Yongguan; Smith, F. Andrew; Wang Youshan; Chen Baodong

    2008-01-01

    In a compartmented cultivation system, white clover (Trifolium repens Linn.) and ryegrass (Lolium perenne L.), with their roots freely intermingled, or separated by 37 μm nylon mesh or plastic board, were grown together in an arsenic (As) contaminated soil. The influence of AM inoculation on plant growth, As uptake, phosphorus (P) nutrition, and plant competitions were investigated. Results showed that both plant species highly depended on mycorrhizas for surviving the As contamination. Mycorrhizal inoculation substantially improved plant P nutrition, and in contrast markedly decreased root to shoot As translocation and shoot As concentrations. It also showed that mycorrhizas affected the competition between the two co-existing plant species, preferentially benefiting the clover plants in term of nutrient acquisition and biomass production. Based on the present study, the role of AM fungi in plant adaptation to As contamination, and their potential use for ecological restoration of As contaminated soils are discussed. - Both white clover and ryegrass highly depend on the mycorrhizal associations for surviving heavy arsenic contamination

  14. Arbuscular mycorrhiza enhanced arsenic resistance of both white clover (Trifolium repens Linn.) and ryegrass (Lolium perenne L.) plants in an arsenic-contaminated soil

    Energy Technology Data Exchange (ETDEWEB)

    Dong Yan; Zhu Yongguan [Department of Soil Environmental Science, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, 18 Shuangqing Road, Beijing 100085 (China); Smith, F. Andrew [Soil and Land Systems, School of Earth and Environmental Sciences, Waite Campus, University of Adelaide, Adelaide, SA 5005 (Australia); Wang Youshan [Institute of Plant Nutrition and Resources, Beijing Academy of Agriculture and Forestry, Beijing 100089 (China); Chen Baodong [Department of Soil Environmental Science, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, 18 Shuangqing Road, Beijing 100085 (China)], E-mail: bdchen@rcees.ac.cn

    2008-09-15

    In a compartmented cultivation system, white clover (Trifolium repens Linn.) and ryegrass (Lolium perenne L.), with their roots freely intermingled, or separated by 37 {mu}m nylon mesh or plastic board, were grown together in an arsenic (As) contaminated soil. The influence of AM inoculation on plant growth, As uptake, phosphorus (P) nutrition, and plant competitions were investigated. Results showed that both plant species highly depended on mycorrhizas for surviving the As contamination. Mycorrhizal inoculation substantially improved plant P nutrition, and in contrast markedly decreased root to shoot As translocation and shoot As concentrations. It also showed that mycorrhizas affected the competition between the two co-existing plant species, preferentially benefiting the clover plants in term of nutrient acquisition and biomass production. Based on the present study, the role of AM fungi in plant adaptation to As contamination, and their potential use for ecological restoration of As contaminated soils are discussed. - Both white clover and ryegrass highly depend on the mycorrhizal associations for surviving heavy arsenic contamination.

  15. Photoaffinity labelling of the active site of the rat glutathione transferases 3-3 and 1-1 and human glutathione transferase A1-1.

    OpenAIRE

    Cooke, R J; Björnestedt, R; Douglas, K T; McKie, J H; King, M D; Coles, B; Ketterer, B; Mannervik, B

    1994-01-01

    The glutathione transferases (GSTs) form a group of enzymes responsible for a wide range of molecular detoxications. The photoaffinity label S-(2-nitro-4-azidophenyl)glutathione was used to study the hydrophobic region of the active site of the rat liver GST 1-1 and 2-2 isoenzymes (class Alpha) as well as the rat class-Mu GST 3-3. Photoaffinity labelling was carried out using a version of S-(2-nitro-4-azidophenyl)glutathione tritiated in the arylazido ring. The labelling occurred with higher ...

  16. Red clover-derived isoflavones and mammographic breast density: a double-blind, randomized, placebo-controlled trial [ISRCTN42940165

    International Nuclear Information System (INIS)

    Atkinson, Charlotte; Warren, Ruth ML; Sala, Evis; Dowsett, Mitch; Dunning, Alison M; Healey, Catherine S; Runswick, Shirley; Day, Nicholas E; Bingham, Sheila A

    2004-01-01

    Isoflavones are hypothesized to protect against breast cancer, but it is not clear whether they act as oestrogens or anti-oestrogens in breast tissue. Our aim was to determine the effects of taking a red clover-derived isoflavone supplement daily for 1 year on mammographic breast density. Effects on oestradiol, follicle-stimulating hormone (FSH), luteinizing hormone (LH), lymphocyte tyrosine kinase activity and menopausal symptoms were also assessed. A total of 205 women (age range 49–65 years) with Wolfe P2 or DY mammographic breast patterns were randomly assigned to receive either a red clover-derived isoflavone tablet (26 mg biochanin A, 16 mg formononetin, 1 mg genistein and 0.5 mg daidzein) or placebo. Change in mammographic breast density, serum oestradiol, FSH, LH, menopausal symptoms and lymphocyte tyrosine kinase activity from baseline to 12 months were assessed. A total of 177 women completed the trial. Mammographic breast density decreased in both groups but the difference between the treatment and placebo was not statistically significant. There was a significant interaction between treatment group and oestrogen receptor (ESR1) PvuII polymorphism for the change in estimated percentage breast density (mean ± standard deviation): TT isoflavone 1.4 ± 12.3% and TT placebo -9.6 ± 14.2%; CT isoflavone -5.2 ± 12.0% and CT placebo -2.8 ± 10.3%; and CC isoflavone -3.4 ± 9.7% and CC placebo -1.1 ± 9.5%. There were no statistically significant treatment effects on oestradiol, FSH, or LH (assessed only in postmenopausal women), or on lymphocyte tyrosine kinase activity. Baseline levels of menopausal symptoms were low, and there were no statistically significant treatment effects on frequency of hot flushes or other menopausal symptoms. In contrast to studies showing that conventional hormone replacement therapies increase mammographic breast density, the isoflavone supplement did not increase mammographic breast density in this population of women

  17. Changes During Growth in Cell Wall Components of Berseem Clover Under Different Cutting Treatments in a Mediterranean Region

    Directory of Open Access Journals (Sweden)

    Giuditta De Santis

    2007-09-01

    Full Text Available Forage digestibility of berseem clover (Trifolium Alexandrinum L. is influenced by plant cell wall composition. This study was conducted to evaluate the effects of different cutting treatments on cell-wall components of the herbage and plant fractions in two Mediterranean berseem genotypes during growth and to examine the relationship between digestibility and cell wall components in these plant fractions. Cutting treatments were initiated at sixth internode elongation (A and early flowering (B and there was an uncut control treatment (C. Spring growth of genotypes of Egyptian (cv. Giza 10 and Italian (cv. Sacromonte origins was harvested in each of two years beginning 196 days after sowing and thereafter every 6 days (twelve harvests in total to measure cell wall components. Neutral detergent fibre (NDF, acid detergent fibre (ADF and acid detergent lignin (ADL concentrations were determined for leaf, stem, and total forage of each cultivar at each harvest. Without defoliation, NDF, ADF and ADL concentrations of herbage increased linearly to a maximum of 528, 375 and 84 g kg-1 DM, respectively, by 220 days after planting, when plant height reached maximum values, then plateaued until 257 days after planting. Plant parts differed in cell-wall concentration, with stems being of higher fibre components than leaves, in the two cutting treatments. Herbage and plant fraction fibre concentrations were negatively correlated with digestibility for all cutting treatments. Defoliation induced a reduction in fibre concentrations and plant height, although a rapid regrowth was observed after cutting, in both treatments A and B. However, delaying defoliation to the early flowering stage (B increased fibre components concentrations both at cutting time and during the growing season and significantly reduced the regrowth height. Conversely, plants cut at the sixth internode elongation (treatment A showed lower concentrations of fibre fractions than initiating

  18. Effect of harvest time of red and white clover silage on chewing activity and particle size distribution in boli, rumen content and faeces in cows.

    Science.gov (United States)

    Kornfelt, L F; Nørgaard, P; Weisbjerg, M R

    2013-06-01

    The study examined the effects of harvest time of red and white clover silage on eating and ruminating activity and particle size distribution in feed boli, rumen content and faeces in cows. The clover crops were harvested at two stages of growth and ensiled in bales. Red clover crops had 36% and 45% NDF in dry matter (DM) at early (ER) and late (LR) harvest, respectively, and the white clover crops had 19% and 29% NDF in DM at the early (EW) and late (LW) harvest, respectively. The silages were fed restrictively (80% of ad libitum intake) twice daily to four rumen cannulated non-lactating Jersey cows (588 ± 52 kg) in a 4 × 4 Latin square design. Jaw movements (JM) were recorded for 96 h continuously. Swallowed boli, rumen mat, rumen fluid and faeces samples were collected, washed in nylon bags (0.01 mm pore size) and freeze-dried before dry sieving through 4.750, 2.360, 1.000, 0.500, 0.212 and 0.106 mm into seven fractions. The length (PL) and width (PW) values of rumen and faeces particles within each fraction were measured by use of image analysis. The eating activity (min/kg DM intake; P rumen mat (P rumen fluid (P rumen mat and faeces, but only one peak (mode 1) for PL values. There was no difference in the mean and mode 1 PW and PL value in rumen mat between the four treatments. The mean PL, mode PL, mode 2 PW and mean PW in faeces were highest for LR (P rumen mat and faeces particles are most likely related to the leaves and the stems/petioles. In conclusion, the mean total chewing activity per kg DM was lowest for the white clover silage and increased for both silages due to later harvest time. The mean particle size in boli was smallest for LR, whereas the mean PL and PW in faeces were highest for the LR.

  19. Assessment of Qualitative and Quantitative Characters of Two Persian Clover Ecotypes Inoculated by Rhizobium leguminosarum biovartrifoli and Pesudomonas putida Bacteria

    Directory of Open Access Journals (Sweden)

    R Azamei

    2016-02-01

    Full Text Available Introduction Over the past decades, world attitude has changed towards the reduction of environmental pollutants. Harmful effects of synthetic fertilizers on environment have been identified. Bio-fertilizers are not harmful to the environment, but also they have favorable effects on plant growth processes. Soil biotechnology can be defined as the study of soil organisms and their metabolic processes which may have positive effects on plant yields. The main goal of this study is to asess the biotechnology fertilizers beneficial effects on soil organisms and their subsequently to maximize the yield. It is also our desire conside the soil quality, hygiene and environmental protection along this process. Among the strain of nitrogen-fixing bacteria, symbiotic bacteria such as rhizobium bacteria are important and essential in planning the sustainable farming systems. Several studies have shown that crop varieties which inoculated with rhizobium and pseudomonas were superior in yield production and performance. Material and Methods An experiment was designed as factorial performed in randomized complete block design (RCBD with three replications in Agricultural Research Center of Golpayegan (Isfahan during 2010 – 2011. the purpose of this study was to evaluate the effects of inoculation of two ecotypes of Persian clover by various strains of Rhizobium leguminosarum. Biovar trifoli bacteria accompanied with plant growth promoting rhizobacteria (PGPR Pseudomonas putida was employed to find certain qualitative and quantitative characteristics of clover yield, The main plots included two local ecotypes of Persian clover; Arak Haft Chin (V1 and Isfahan Haft Chin (V2, the subplots included inoculation by two strain of Rhizobium; Rb-3, Rb-13 and one strain of Pseudomonas; PS -168.4 cuts were performed during the experiment and 60 kg/ha seed was used for cultivation based on local knowledge. According to recommendations of the Institute of Soil and Water

  20. Nitrogen-doped two-dimensional porous carbon sheets derived from clover biomass for high performance supercapacitors

    Science.gov (United States)

    Wang, Cunjing; Wu, Dapeng; Wang, Hongju; Gao, Zhiyong; Xu, Fang; Jiang, Kai

    2017-09-01

    Highly porous carbon sheets were prepared from fresh clover stems under air atmosphere via a facile potassium chloride salt-sealing technique, which not only avoids using the high cost inert gas protection but also spontaneously introduce multi-level porosity into the carbon structure taking advantage of the trace of oxygen in the molten salt system. The as-obtained porous carbon sheets possess high specific surface area of 2244 m2 g-1 and interconnected hierarchical pore structures from micro-to macro-scale, which provide abundant storage active sites and fast ion diffusion channels. In addition, the spontaneously formed N (2.55 at%) and O (6.94 at%) doping sites not only improve the electron conductivity of the electrode but also enhance the specific capacitance by introducing pseudocapacitance. When employed as supercapacitor electrodes, a high specific capacitance of 436 F g-1 at 1 A g-1 and an excellent rate capacity with capacitance remaining 290 F g-1 at 50 A g-1 are demonstrated. Furthermore, the assembled symmetric supercapacitor delivers a high specific capacitance of 420 F g-1 at 0.5 A g-1, excellent energy density of 58.4 Wh kg-1 and good cycling stability which retains 99.4% of the initial capacitance at 5 A g-1 after 30,000 cycles.

  1. Interannual variation in nitrous oxide emissions from perennial ryegrass/white clover grassland used for dairy production.

    Science.gov (United States)

    Burchill, William; Li, Dejun; Lanigan, Gary J; Williams, Micheal; Humphreys, James

    2014-10-01

    Nitrous oxide (N2 O) emissions are subject to intra- and interannual variation due to changes in weather and management. This creates significant uncertainties when quantifying estimates of annual N2 O emissions from grazed grasslands. Despite these uncertainties, the majority of studies are short-term in nature (Nitrous oxide emissions were measured from fertilized and grazed perennial ryegrass/white clover grassland (WC) and from perennial ryegrass plots that were not grazed and did not receive N input (GB), over 4 years from 2008 to 2012 in Ireland (52°51'N, 08°21'W). The annual N2 O-N emissions (kg ha(-1); mean ± SE) ranged from 4.4 ± 0.2 to 34.4 ± 5.5 from WC and from 1.7 ± 0.8 to 6.3 ± 1.2 from GB. Interannual variation in N2 O emissions was attributed to differences in annual rainfall, monthly (December) soil temperatures and variation in N input. Such substantial interannual variation in N2 O emissions highlights the need for long-term studies of emissions from managed pastoral systems. © 2014 The Authors. Global Change Biology Published by John Wiley & Sons Ltd.

  2. Nitrogen Fertilizer Sources and Application Timing Affects Wheat and Inter-Seeded Red Clover Yields on Claypan Soils

    Directory of Open Access Journals (Sweden)

    Kelly A. Nelson

    2014-11-01

    Full Text Available Controlled-release N fertilizer, such as polymer-coated urea (PCU, may be a fall N management option for wheat (Triticum aestivum L. grown in poorly-drained claypan soils. Field research evaluated (1 urea release from fall-applied PCU in 2006 and 2007; (2 broadcast fall-spring split (25%:75% of N sources; and (3 a single fall (100% application of PCU, urea, urea plus NBPT (N-(n-butyl thiophosphoric triamide] (U + NBPT, ammonium nitrate (AN, or urea ammonium nitrate (UAN at 0, 56, 84, and 112 kg·N·ha−1 on wheat yield, wheat biomass, N uptake by wheat, and frost-seeded red clover (FSC (Trifolium pratense L. forage yield (2004–2007. PCU applied in fall released less than 30% urea by February. Urea released from PCU by harvest was 60% and 85% in 2006 and 2007, respectively. In poorly-drained soils, wheat yields ranked PCU > AN > U + NBPT > urea ≥ UAN over the rates evaluated for fall-only application. PCU was a viable fall-applied N source, with yields similar to or greater than urea or U + NBPT split-applied. Split-N applications of AN, urea, UAN, and U + NBPT generally resulted in greater wheat yields than a fall application. Enhanced efficiency fertilizers provide farmers with flexible options for maintaining high yielding production systems.

  3. Carcass parameters and meat quality in meat-goat kids finished on chicory, birdsfoot trefoil, or red clover pastures.

    Science.gov (United States)

    Turner, K E; Cassida, K A; Zerby, H N; Brown, M A

    2015-07-01

    This study was conducted during the 2009 and 2010 grazing seasons to assess carcass parameters and chevon (goat meat) quality when meat-goat kids (n=72) were finished on pastures of red clover (Trifolium pratense L.; RCL), birdsfoot trefoil (Lotus corniculatus L.; BFT), or chicory (Cichorium intybus L.; CHIC). Final live weight (Pgoats were finished on RCL compared to CHIC with BFT being intermediate. Ribeye area, backfat thickness, body wall thickness, internal fat score, and leg score were not different (P>0.10) among treatments when adjusted for the covariate of carcass weight. Finishing meat-goat kids on RCL, BFT, or CHIC impacted concentrations of fatty acids (FAs) 18:1 trans-10, 18:1 cis-11, 18:2, 18:3 polyunsaturated fatty acids (PUFAs), omega-6, omega-3, and PUFA:saturated fatty acid ratio in longissimus lumborum samples. Finishing meat-goat kids on CHIC, RCL, or BFT pastures produced carcass weights acceptable for most ethnic markets in the USA. Published by Elsevier Ltd.

  4. Arbuscular mycorrhizal fungi can decrease the uptake of uranium by subterranean clover grown at high levels of uranium in soil

    International Nuclear Information System (INIS)

    Rufyikiri, Gervais; Huysmans, Lien; Wannijn, Jean; Hees, May van; Leyval, Corinne; Jakobsen, Iver

    2004-01-01

    Subterranean clover inoculated or not with the arbuscular mycorrhizal (AM) fungus Glomus intraradices was grown on soil containing six levels of 238 U in the range 0-87 mg kg -1 . Increasing U concentration in soil enhanced the U concentration in roots and shoots of both mycorrhizal and nonmycorrhizal plants but had no significant effects on plant dry matter production or root AM colonization. Mycorrhizas increased the shoot dry matter and P concentration in roots and shoots, while in most cases, it decreased the Ca, Mg and K concentrations in plants. The AM fungus influenced U concentration in plants only in the treatment receiving 87 mg U kg -1 soil. In this case, U concentration in shoots of nonmycorrhizal plants was 1.7 times that of shoots of mycorrhizal plants. These results suggested that mycorrhizal fungi can limit U accumulation by plants exposed to high levels of U in soil. - Plant mycorrhization may decrease U concentration in shoots of plants grown at high level of U in soil

  5. A novel plant glutathione S-transferase/peroxidase suppresses Bax lethality in yeast

    DEFF Research Database (Denmark)

    Kampranis, S C; Damianova, R; Atallah, M

    2000-01-01

    The mammalian inducer of apoptosis Bax is lethal when expressed in yeast and plant cells. To identify potential inhibitors of Bax in plants we transformed yeast cells expressing Bax with a tomato cDNA library and we selected for cells surviving after the induction of Bax. This genetic screen allows...... for the identification of plant genes, which inhibit either directly or indirectly the lethal phenotype of Bax. Using this method a number of cDNA clones were isolated, the more potent of which encodes a protein homologous to the class theta glutathione S-transferases. This Bax-inhibiting (BI) protein was expressed...... in Escherichia coli and found to possess glutathione S-transferase (GST) and weak glutathione peroxidase (GPX) activity. Expression of Bax in yeast decreases the intracellular levels of total glutathione, causes a substantial reduction of total cellular phospholipids, diminishes the mitochondrial membrane...

  6. Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity

    DEFF Research Database (Denmark)

    Pedersen, Johannes W; Bennett, Eric P; Schjoldager, Katrine T-B G

    2011-01-01

    UDP-GalNAc:polypeptide a-N-acetylgalactosaminyltransferases (GalNAc-Ts) constitute a family of up to 20 transferases that initiate mucin-type O-glycosylation. The transferases are structurally composed of catalytic and lectin domains. Two modes have been identified for the selection...... of glycosylation sites by GalNAc-Ts: confined sequence recognition by the catalytic domain alone, and concerted recognition of acceptor sites and adjacent GalNAc-glycosylated sites by the catalytic and lectin domains, respectively. Thus far, only the catalytic domain has been shown to have peptide sequence...... on sequences of mucins MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC7 as well as a random glycopeptide bead library, we examined the binding properties of four different lectin domains. The lectin domains of GalNAc-T1, -T2, -T3, and -T4 bound different subsets of small glycopeptides. These results indicate...

  7. Development of isoform-specific sensors of polypeptide GalNAc-transferase activity

    DEFF Research Database (Denmark)

    Song, Lina; Bachert, Collin; Schjoldager, Katrine T

    2014-01-01

    sequence influenced their activity and required modification, which we carried out based on previous in vitro work. Significantly, the modified T2 and T3 sensors were activated only in cells lacking their corresponding isozymes. Thus, we have developed T2- and T3-specific sensors that will be valuable......Humans express up to 20 isoforms of GalNAc-transferase (herein T1-T20) that localize to the Golgi apparatus and initiate O-glycosylation. Regulation of this enzyme family affects a vast array of proteins transiting the secretory pathway and diseases arise upon misregulation of specific isoforms....... Surprisingly, molecular probes to monitor GalNAc-transferase activity are lacking and there exist no effective global or isoform-specific inhibitors. Here we describe the development of T2- and T3-isoform specific fluorescence sensors that traffic in the secretory pathway. Each sensor yielded little signal...

  8. Caractérisation biochimique et fonctionnelle de glutathion-S-transferases (GSTs) chez Phanerochaete chrysosporium

    OpenAIRE

    Anak Ngadin , Andrew

    2011-01-01

    Phanerochaete chrysosporium is a ligninolytic fungus widely studied because of its capacities to degrade wood and xenobiotics through an extracellular enzymatic system. Its genome has been sequenced and has provided researchers with a complete inventory of the predicted proteins produced by this organism. This has allowed the description of many protein superfamilies. Among them, Glutathione S-transferases (GSTs) constitute a complex and widespread superfamily classified as enzymes of seconda...

  9. Imidazopyridine and Pyrazolopiperidine Derivatives as Novel Inhibitors of Serine Palmitoyl Transferase.

    Science.gov (United States)

    Genin, Michael J; Gonzalez Valcarcel, Isabel C; Holloway, William G; Lamar, Jason; Mosior, Marian; Hawkins, Eric; Estridge, Thomas; Weidner, Jeffrey; Seng, Thomas; Yurek, David; Adams, Lisa A; Weller, Jennifer; Reynolds, Vincent L; Brozinick, Joseph T

    2016-06-23

    To develop novel treatments for type 2 diabetes and dyslipidemia, we pursued inhibitors of serine palmitoyl transferase (SPT). To this end compounds 1 and 2 were developed as potent SPT inhibitors in vitro. 1 and 2 reduce plasma ceramides in rodents, have a slight trend toward enhanced insulin sensitization in DIO mice, and reduce triglycerides and raise HDL in cholesterol/cholic acid fed rats. Unfortunately these molecules cause a gastric enteropathy after chronic dosing in rats.

  10. The phosphopantetheinyl transferases: catalysis of a post-translational modification crucial for life

    DEFF Research Database (Denmark)

    Beld, Joris; Sonnenschein, Eva; Vickery, Christopher R.

    2013-01-01

    Covering: up to 2013 Although holo-acyl carrier protein synthase, AcpS, a phosphopantetheinyl transferase (PPTase), was characterized in the 1960s, it was not until the publication of the landmark paper by Lambalot et al. in 1996 that PPTases garnered wide-spread attention being classified...... knowledge on this class of enzymes that post-translationally install a 4′-phosphopantetheine arm on various carrier proteins....

  11. A 4'-phosphopantetheinyl transferase mediates non-ribosomal peptide synthetase activation in Aspergillus fumigatus.

    Science.gov (United States)

    Neville, Claire; Murphy, Alan; Kavanagh, Kevin; Doyle, Sean

    2005-04-01

    Aspergillus fumigatus is a significant human pathogen. Non-ribosomal peptide (NRP) synthesis is thought to be responsible for a significant proportion of toxin and siderophore production in the organism. Furthermore, it has been shown that 4'-phosphopantetheinylation is required for the activation of key enzymes involved in non-ribosomal peptide synthesis in other species. Here we report the cloning, recombinant expression and functional characterisation of a 4'-phosphopantetheinyl transferase from A. fumigatus and the identification of an atypical NRP synthetase (Afpes1), spanning 14.3 kb. Phylogenetic analysis has shown that the NRP synthetase exhibits greatest identity to NRP synthetases from Metarhizium anisolpiae (PesA) and Alternaria brassicae (AbrePsy1). Northern hybridisation and RT-PCR analysis have confirmed that both genes are expressed in A. fumigatus. A 120 kDa fragment of the A. fumigatus NRP synthetase, containing a putative thiolation domain, was cloned and expressed in the baculovirus expression system. Detection of a 4'-phosphopantetheinylated peptide (SFSAMK) from this protein, by MALDI-TOF mass spectrometric analysis after coincubation of the 4'-phosphopantetheinyl transferase with the recombinant NRP synthetase fragment and acetyl CoA, confirms that it is competent to play a role in NRP synthetase activation in A. fumigatus. The 4'-phosphopantetheinyl transferase also activates, by 4'-phosphopantetheinylation, recombinant alpha-aminoadipate reductase (Lys2p) from Candida albicans, a key enzyme involved in lysine biosynthesis.

  12. Interactions of photoactive DNAs with terminal deoxynucleotidyl transferase: Identification of peptides in the DNA binding domain

    International Nuclear Information System (INIS)

    Farrar, Y.J.K.; Evans, R.K.; Beach, C.M.; Coleman, M.S.

    1991-01-01

    Terminal deoxynucleotidyl transferase (terminal transferase) was specifically modified in the DNA binding site by a photoactive DNA substrate (hetero-40-mer duplex containing eight 5-azido-dUMP residues at one 3' end). Under optimal photolabeling conditions, 27-40% of the DNA was covalently cross-linked to terminal transferase. The specificity of the DNA and protein interaction was demonstrated by protection of photolabeling at the DNA binding domain with natural DNA substrates. In order to recover high yields of modified peptides from limited amounts of starting material, protein modified with 32 P-labeled photoactive DNA and digested with trypsin was extracted 4 times with phenol followed by gel filtration chromatography. All peptides not cross-linked to DNA were extracted into the phenol phase while the photolyzed DNA and the covalently cross-linked peptides remained in the aqueous phase. The 32 P-containing peptide-DNA fraction was subjected to amino acid sequence analysis. Two sequences, Asp 221 -Lys 231 (peptide B8) and Cys 234 -Lys 249 (peptide B10), present in similar yield, were identified. Structure predictions placed the two peptides in an α-helical array of 39 angstrom which would accommodate a DNA helix span of 11 nucleotides. These peptides share sequence similarity with a region in DNA polymerase β that has been implicated in the binding of DNA template

  13. Glutathione S - transferases class Pi and Mi and their significance in oncology

    Directory of Open Access Journals (Sweden)

    Zofia Marchewka

    2017-06-01

    Full Text Available In this article the current data, which shows that glutathione S-transferases (GST class Pi and Mi are interesting and promising biomarkers in acute and chronic inflammatory processes as well as in the oncology, were presented based on the review of the latest experimental and clinical studies. The article shows their characteristics, functions and participation (direct - GST Pi, indirect - GST Mi in the regulation of signaling pathways of JNK kinases, which are involved in cell differentiation. Overexpression of glutathione S-transferases class Pi and Mi in many cancer cells plays a key role in cancer treatment, making them resistant to chemotherapy. GST isoenzymes are involved in the metabolism of various types of xenobiotics and endogenous substrates, so their altered expression in cancer tissues as well as in serum and urine could be an important potential marker of the cancer and an indicator of oxidative stress. The study shows the role of glutathione S-transferases in redox homeostasis of tumor cells and in the mechanism of resistance to anticancer drugs.

  14. Glutathione S - transferases class Pi and Mi and their significance in oncology.

    Science.gov (United States)

    Marchewka, Zofia; Piwowar, Agnieszka; Ruzik, Sylwia; Długosz, Anna

    2017-06-19

    In this article the current data, which shows that glutathione S-transferases (GST) class Pi and Mi are interesting and promising biomarkers in acute and chronic inflammatory processes as well as in the oncology, were presented based on the review of the latest experimental and clinical studies. The article shows their characteristics, functions and participation (direct - GST Pi, indirect - GST Mi) in the regulation of signaling pathways of JNK kinases, which are involved in cell differentiation. Overexpression of glutathione S-transferases class Pi and Mi in many cancer cells plays a key role in cancer treatment, making them resistant to chemotherapy. GST isoenzymes are involved in the metabolism of various types of xenobiotics and endogenous substrates, so their altered expression in cancer tissues as well as in serum and urine could be an important potential marker of the cancer and an indicator of oxidative stress. The study shows the role of glutathione S-transferases in redox homeostasis of tumor cells and in the mechanism of resistance to anticancer drugs.

  15. Characterization of Affinity-Purified Isoforms of Acinetobacter calcoaceticus Y1 Glutathione Transferases

    Directory of Open Access Journals (Sweden)

    Chin-Soon Chee

    2014-01-01

    Full Text Available Glutathione transferases (GST were purified from locally isolated bacteria, Acinetobacter calcoaceticus Y1, by glutathione-affinity chromatography and anion exchange, and their substrate specificities were investigated. SDS-polyacrylamide gel electrophoresis revealed that the purified GST resolved into a single band with a molecular weight (MW of 23 kDa. 2-dimensional (2-D gel electrophoresis showed the presence of two isoforms, GST1 (pI 4.5 and GST2 (pI 6.2 with identical MW. GST1 was reactive towards ethacrynic acid, hydrogen peroxide, 1-chloro-2,4-dinitrobenzene, and trans,trans-hepta-2,4-dienal while GST2 was active towards all substrates except hydrogen peroxide. This demonstrated that GST1 possessed peroxidase activity which was absent in GST2. This study also showed that only GST2 was able to conjugate GSH to isoproturon, a herbicide. GST1 and GST2 were suggested to be similar to F0KLY9 (putative glutathione S-transferase and F0KKB0 (glutathione S-transferase III of Acinetobacter calcoaceticus strain PHEA-2, respectively.

  16. Characterization of Affinity-Purified Isoforms of Acinetobacter calcoaceticus Y1 Glutathione Transferases

    Science.gov (United States)

    Chee, Chin-Soon; Tan, Irene Kit-Ping; Alias, Zazali

    2014-01-01

    Glutathione transferases (GST) were purified from locally isolated bacteria, Acinetobacter calcoaceticus Y1, by glutathione-affinity chromatography and anion exchange, and their substrate specificities were investigated. SDS-polyacrylamide gel electrophoresis revealed that the purified GST resolved into a single band with a molecular weight (MW) of 23 kDa. 2-dimensional (2-D) gel electrophoresis showed the presence of two isoforms, GST1 (pI 4.5) and GST2 (pI 6.2) with identical MW. GST1 was reactive towards ethacrynic acid, hydrogen peroxide, 1-chloro-2,4-dinitrobenzene, and trans,trans-hepta-2,4-dienal while GST2 was active towards all substrates except hydrogen peroxide. This demonstrated that GST1 possessed peroxidase activity which was absent in GST2. This study also showed that only GST2 was able to conjugate GSH to isoproturon, a herbicide. GST1 and GST2 were suggested to be similar to F0KLY9 (putative glutathione S-transferase) and F0KKB0 (glutathione S-transferase III) of Acinetobacter calcoaceticus strain PHEA-2, respectively. PMID:24892084

  17. Effect of substituting fresh-cut perennial ryegrass with fresh-cut white clover on bovine milk fatty acid profile.

    Science.gov (United States)

    Stergiadis, Sokratis; Hynes, Deborah N; Thomson, Anna L; Kliem, Kirsty E; Berlitz, Carolina Gb; Günal, Mevlüt; Yan, Tianhai

    2018-03-06

    Including forage legumes in dairy systems can help address increasing environmental/economic concerns about perennial ryegrass monoculture pastures. This work investigated the effect of substituting fresh-cut grass with increasing quantities of fresh-cut white clover (WC) on milk fatty acid (FA) profile and transfer efficiency of dietary linoleic (LA) and α-linolenic (ALNA) acids to milk fat. Three groups of three crossbred dairy cows were used in a 3 × 3 crossover design. Dietary treatments were 0 g kg -1 WC + 600 g kg -1 grass, 200 g kg -1 WC + 400 g kg -1 grass, and 400 g kg -1 WC + 200 g kg -1 grass. All treatments were supplemented with 400 g kg -1 concentrates on a dry matter basis. Cows had a 19-day adaptation period to the experimental diet before a 6-day measurement period in individual tie stalls. Increasing dietary WC did not affect dry matter intake, milk yield or milk concentrations of fat, protein or lactose. Milk polyunsaturated FA concentrations (total n-3, total n-6, LA and ALNA) and transfer efficiency of LA and ALNA were increased with increasing dietary WC supply. Inclusion of WC in pastures may increase concentrations of nutritionally beneficial FA, without influencing milk yield and basic composition, but any implications on human health cannot be drawn. © 2018 The Authors. Journal of the Science of Food and Agriculture published by JohnWiley & Sons Ltd on behalf of Society of Chemical Industry. © 2018 The Authors. Journal of the Science of Food and Agriculture published by JohnWiley & Sons Ltd on behalf of Society of Chemical Industry.

  18. Comparison of buckwheat, red clover, and purple tansy as potential surrogate plants for use in semi-field pesticide risk assessments with Bombus impatiens

    Directory of Open Access Journals (Sweden)

    Angela E. Gradish

    2016-07-01

    Full Text Available Background. Bumble bees (Bombus spp. are important wild and managed pollinators. There is increased interest in incorporating data on bumble bees into risk assessments for pesticides, but standardized methods for assessing hazards of pesticides in semi-field and field settings have not yet been established for bumble bees. During semi-field studies, colonies are caged with pesticide-treated flowering surrogate plants, which must be attractive to foragers to ensure colony exposure to the test compound, and must produce an ample nectar and pollen to sustain colonies during testing. However, it is not known which plant(s are suitable for use in semi-field studies with bumble bees. Materials and Methods. We compared B. impatiens foraging activity and colony development on small plots of flowering buckwheat (Fagopyrum esculentum, var. common, red clover (Trifolium pratense, and purple tansy (Phacelia tanacetifolia under semi-field conditions to assess their suitability as surrogate plants for pesticide risk assessment studies with bumble bees. We also compared the growth characteristics and input requirements of each plant type. Results. All three plant types generally established and grew well. Red clover and purple tansy experienced significant weed pressure and/or insect pest damage. In contrast, pest pressure was extremely low in buckwheat. Overall, B. impatiens foraging activity was significantly greater on buckwheat plots than red clover or purple tansy, but plant type had no effect on number of individuals produced per colony or colony weight. Discussion. Because of the consistently high foraging activity and successful colony development observed on buckwheat plots, combined with its favourable growth characteristics and low maintenance requirements, we recommend buckwheat as a surrogate plant for use in semi-field pesticide toxicity assessments with B. impatiens.

  19. Discussion of a Well-Designed Clinical Trial Which Did Not Demonstrate Effectiveness: UIC Center for Botanical Dietary Supplements Research Study of Black Cohosh and Red Clover

    Science.gov (United States)

    Shulman, Lee P.; Banuvar, Suzanne; Fong, Harry H. S.; Farnsworth, Norman R.

    2016-01-01

    The performance of a clinical trial for pharmaceutical agents is usually undertaken only after there is likely benefit demonstrated from the use of the putative agent. The consideration of botanical products as pharmaceutical agents must similarly go through a rigorous evaluation process. The present work reviews the recently published Phase II study evaluating the effectiveness of black cohosh and red clover in a randomized trial with conjugated equine estradiol/medroxyprogesterone acetate and placebo for the treatment of menopausal symptoms. We analyze the possible reasons why this study failed to show benefit for either botanical product in reducing menopause-related vasomotor symptoms. PMID:21034798

  20. Identification of an extensive gene cluster among a family of PPOs in Trifolium pratense L. (red clover using a large insert BAC library

    Directory of Open Access Journals (Sweden)

    Thomas Ann

    2009-07-01

    Full Text Available Abstract Background Polyphenol oxidase (PPO activity in plants is a trait with potential economic, agricultural and environmental impact. In relation to the food industry, PPO-induced browning causes unacceptable discolouration in fruit and vegetables: from an agriculture perspective, PPO can protect plants against pathogens and environmental stress, improve ruminant growth by increasing nitrogen absorption and decreasing nitrogen loss to the environment through the animal's urine. The high PPO legume, red clover, has a significant economic and environmental role in sustaining low-input organic and conventional farms. Molecular markers for a range of important agricultural traits are being developed for red clover and improved knowledge of PPO genes and their structure will facilitate molecular breeding. Results A bacterial artificial chromosome (BAC library comprising 26,016 BAC clones with an average 135 Kb insert size, was constructed from Trifolium pratense L. (red clover, a diploid legume with a haploid genome size of 440–637 Mb. Library coverage of 6–8 genome equivalents ensured good representation of genes: the library was screened for polyphenol oxidase (PPO genes. Two single copy PPO genes, PPO4 and PPO5, were identified to add to a family of three, previously reported, paralogous genes (PPO1–PPO3. Multiple PPO1 copies were identified and characterised revealing a subfamily comprising three variants PPO1/2, PPO1/4 and PPO1/5. Six PPO genes clustered within the genome: four separate BAC clones could be assembled onto a predicted 190–510 Kb single BAC contig. Conclusion A PPO gene family in red clover resides as a cluster of at least 6 genes. Three of these genes have high homology, suggesting a more recent evolutionary event. This PPO cluster covers a longer region of the genome than clusters detected in rice or previously reported in tomato. Full-length coding sequences from PPO4, PPO5, PPO1/5 and PPO1/4 will facilitate

  1. The effects of PPO activity on the proteome of ingested red clover and implications for improving the nutrition of grazing cattle.

    Science.gov (United States)

    Hart, E H; Onime, L A; Davies, T E; Morphew, R M; Kingston-Smith, A H

    2016-06-01

    Increasing the rumen-stable protein content of feed would lead to improved nitrogen utilisation in cattle, and less nitrogenous waste. Red clover (Trifolium pratense L.) is a high protein ruminant feed containing high polyphenol oxidase (PPO) activity. PPO mediated protein-quinone binding has been linked to protecting plant proteins from proteolysis. To explore the mechanism underlying the effect of PPO on protein protection in fresh forage feeds, proteomic components of feed down-boli produced from wild-type red clover and a low PPO mutant, at point of ingestion and after 4h in vitro incubation with rumen inoculum were analysed. Significant differences in proteomic profiles between wild-type and mutant red clover were determined after 4h incubation, with over 50% less spots in mutant than wild-type proteomes, indicating decreased proteolysis in the latter. Protein identifications revealed preferentially retained proteins localised within the chloroplast, suggesting that PPO mediated protection in the wild-type operates due to the proximity of target proteins to the enzyme and substrates, either diffusing into this compartment from the vacuole or are present in the chloroplast. This increased understanding of protein targets of PPO indicates that wider exploitation of the trait could contribute to increased protein use efficiency in grazing cattle. One of the main challenges for sustainable livestock farming is improving capture of dietary nitrogen by ruminants. Typically up to 70% of ingested protein-N is excreted representing a loss of productivity potential and a serious environmental problem in terms of nitrogenous pollution of lands and water. Identification of key characteristics of rumen-protected protein will deliver target traits for selection in forage breeding programmes. The chloroplastic enzyme PPO catalyzes the oxidation of phenols to quinones, which react with protein. Little is currently known about the intracellular protein targets of the products

  2. Forage fiber effects on particle size reduction, ruminal stratification, and selective retention in heifers fed highly digestible grass/clover silages.

    Science.gov (United States)

    Schulze, A K S; Weisbjerg, M R; Storm, A C; Nørgaard, P

    2014-06-01

    The objective of this study was to investigate the effect of NDF content in highly digestible grass/clover silage on particle size reduction, ruminal stratification, and selective retention in dairy heifers. The reduction in particle size from feed to feces was evaluated and related to feed intake, chewing activity, and apparent digestibility. Four grass/clover harvests (Mixtures of Lolium perenne, Trifolium pratense, and Trifolium repens) were performed from early May to late August at different maturities, at different regrowth stages, and with different clover proportions, resulting in silages with NDF contents of 312, 360, 371, and 446 g/kg DM, respectively, and decreasing NDF digestibility with greater NDF content. Four rumen-fistulated dairy heifers were fed silage at 90% of ad libitum level as the only feed source in a 4 × 4 Latin square design. Silage, ingested feed boluses, medial and ventral ruminal digesta, and feces samples were washed with neutral detergent in nylon bags of 10-μm pore size, freeze dried, and divided into small (1 mm) particles by dry-sieving. Chewing activity, rumen pool size, and apparent digestibility were measured. Intake of NDF increased linearly from 2.3 to 2.8 kg/d with greater NDF content of forages (P = 0.01), but silages were exposed to similar eating time (P = 0.55) and rumination time per kg NDF (P = 0.35). No linear effect of NDF content was found on proportion of LP in ingested feed boluses (P = 0.31), medial rumen digesta (P = 0.95), ventral rumen digesta (P = 0.84), and feces (P = 0.09). Greater proportions of DM (P ruminal digesta compared with ventral rumen, and differences in DM proportion increased with greater NDF content (P = 0.02). Particle size distributions were similar for digesta from the medial and ventral rumen regardless of NDF content of the silages (P > 0.13). The LP proportion was >30% of particles in the ventral and medial rumen, whereas in the feces, the LP proportion was content of the silages

  3. Catalysis of Silver catfish Major Hepatic Glutathione Transferase proceeds via rapid equilibrium sequential random Mechanism

    Directory of Open Access Journals (Sweden)

    Ayodele O. Kolawole

    Full Text Available Fish hepatic glutathione transferases are connected with the elimination of intracellular pollutants and detoxification of organic micro-pollutants in their aquatic ecosystem. The two-substrate steady state kinetic mechanism of Silver catfish (Synodontis eupterus major hepatic glutathione transferases purified to apparent homogeneity was explored. The enzyme was dimeric enzyme with a monomeric size of 25.6 kDa. Initial-velocity studies and Product inhibition patterns by methyl glutathione and chloride with respect to GSH-CDNB; GSH-ρ-nitrophenylacetate; and GSH-Ethacrynic acid all conforms to a rapid equilibrium sequential random Bi Bi kinetic mechanism rather than steady state sequential random Bi Bi kinetic. α was 2.96 ± 0.35 for the model. The pH profile of Vmax/KM (with saturating 1-chloro-2,4-dinitrobenzene and variable GSH concentrations showed apparent pKa value of 6.88 and 9.86. Inhibition studies as a function of inhibitor concentration show that the enzyme is a homodimer and near neutral GST. The enzyme poorly conjugates 4-hydroxylnonenal and cumene hydroperoxide and may not be involved in oxidative stress protection. The seGST is unique and overwhelmingly shows characteristics similar to those of homodimeric class Pi GSTs, as was indicated by its kinetic mechanism, substrate specificity and inhibition studies. The rate- limiting step, probably the product release, of the reaction is viscosity-dependent and is consequential if macro-viscosogen or micro-viscosogen. Keywords: Silver catfish, Glutathione transferase, Steady-state, Kinetic mechanism, Inhibition

  4. Characterization of glutathione transferases involved in the pathogenicity of Alternaria brassicicola.

    Science.gov (United States)

    Calmes, Benoit; Morel-Rouhier, Mélanie; Bataillé-Simoneau, Nelly; Gelhaye, Eric; Guillemette, Thomas; Simoneau, Philippe

    2015-06-18

    Glutathione transferases (GSTs) represent an extended family of multifunctional proteins involved in detoxification processes and tolerance to oxidative stress. We thus anticipated that some GSTs could play an essential role in the protection of fungal necrotrophs against plant-derived toxic metabolites and reactive oxygen species that accumulate at the host-pathogen interface during infection. Mining the genome of the necrotrophic Brassica pathogen Alternaria brassicicola for glutathione transferase revealed 23 sequences, 17 of which could be clustered into the main classes previously defined for fungal GSTs and six were 'orphans'. Five isothiocyanate-inducible GSTs from five different classes were more thoroughly investigated. Analysis of their catalytic properties revealed that two GSTs, belonging to the GSTFuA and GTT1 classes, exhibited GSH transferase activity with isothiocyanates (ITC) and peroxidase activity with cumene hydroperoxide, respectively. Mutant deficient for these two GSTs were however neither more susceptible to ITC nor less aggressive than the wild-type parental strain. By contrast mutants deficient for two other GSTs, belonging to the Ure2pB and GSTO classes, were distinguished by their hyper-susceptibility to ITC and low aggressiveness against Brassica oleracea. In particular AbGSTO1 could participate in cell tolerance to ITC due to its glutathione-dependent thioltransferase activity. The fifth ITC-inducible GST belonged to the MAPEG class and although it was not possible to produce the soluble active form of this protein in a bacterial expression system, the corresponding deficient mutant failed to develop normal symptoms on host plant tissues. Among the five ITC-inducible GSTs analyzed in this study, three were found essential for full aggressiveness of A. brassicicola on host plant. This, to our knowledge is the first evidence that GSTs might be essential virulence factors for fungal necrotrophs.

  5. Glutathione Transferase from Trichoderma virens Enhances Cadmium Tolerance without Enhancing Its Accumulation in Transgenic Nicotiana tabacum

    Science.gov (United States)

    Dixit, Prachy; Mukherjee, Prasun K.; Ramachandran, V.; Eapen, Susan

    2011-01-01

    Background Cadmium (Cd) is a major heavy metal pollutant which is highly toxic to plants and animals. Vast agricultural areas worldwide are contaminated with Cd. Plants take up Cd and through the food chain it reaches humans and causes toxicity. It is ideal to develop plants tolerant to Cd, without enhanced accumulation in the edible parts for human consumption. Glutathione transferases (GST) are a family of multifunctional enzymes known to have important roles in combating oxidative stresses induced by various heavy metals including Cd. Some GSTs are also known to function as glutathione peroxidases. Overexpression/heterologous expression of GSTs is expected to result in plants tolerant to heavy metals such as Cd. Results Here, we report cloning of a glutathione transferase gene from Trichoderma virens, a biocontrol fungus and introducing it into Nicotiana tabacum plants by Agrobacterium-mediated gene transfer. Transgenic nature of the plants was confirmed by Southern blot hybridization and expression by reverse transcription PCR. Transgene (TvGST) showed single gene Mendelian inheritance. When transgenic plants expressing TvGST gene were exposed to different concentrations of Cd, they were found to be more tolerant compared to wild type plants, with transgenic plants showing lower levels of lipid peroxidation. Levels of different antioxidant enzymes such as glutathione transferase, superoxide dismutase, ascorbate peroxidase, guiacol peroxidase and catalase showed enhanced levels in transgenic plants expressing TvGST compared to control plants, when exposed to Cd. Cadmium accumulation in the plant biomass in transgenic plants were similar or lower than wild-type plants. Conclusion The results of the present study suggest that transgenic tobacco plants expressing a Trichoderma virens GST are more tolerant to Cd, without enhancing its accumulation in the plant biomass. It should be possible to extend the present results to crop plants for developing Cd tolerance and

  6. Glutathione transferase from Trichoderma virens enhances cadmium tolerance without enhancing its accumulation in transgenic Nicotiana tabacum.

    Directory of Open Access Journals (Sweden)

    Prachy Dixit

    Full Text Available BACKGROUND: Cadmium (Cd is a major heavy metal pollutant which is highly toxic to plants and animals. Vast agricultural areas worldwide are contaminated with Cd. Plants take up Cd and through the food chain it reaches humans and causes toxicity. It is ideal to develop plants tolerant to Cd, without enhanced accumulation in the edible parts for human consumption. Glutathione transferases (GST are a family of multifunctional enzymes known to have important roles in combating oxidative stresses induced by various heavy metals including Cd. Some GSTs are also known to function as glutathione peroxidases. Overexpression/heterologous expression of GSTs is expected to result in plants tolerant to heavy metals such as Cd. RESULTS: Here, we report cloning of a glutathione transferase gene from Trichoderma virens, a biocontrol fungus and introducing it into Nicotiana tabacum plants by Agrobacterium-mediated gene transfer. Transgenic nature of the plants was confirmed by Southern blot hybridization and expression by reverse transcription PCR. Transgene (TvGST showed single gene Mendelian inheritance. When transgenic plants expressing TvGST gene were exposed to different concentrations of Cd, they were found to be more tolerant compared to wild type plants, with transgenic plants showing lower levels of lipid peroxidation. Levels of different antioxidant enzymes such as glutathione transferase, superoxide dismutase, ascorbate peroxidase, guiacol peroxidase and catalase showed enhanced levels in transgenic plants expressing TvGST compared to control plants, when exposed to Cd. Cadmium accumulation in the plant biomass in transgenic plants were similar or lower than wild-type plants. CONCLUSION: The results of the present study suggest that transgenic tobacco plants expressing a Trichoderma virens GST are more tolerant to Cd, without enhancing its accumulation in the plant biomass. It should be possible to extend the present results to crop plants for

  7. The dyad palindromic glutathione transferase P enhancer binds multiple factors including AP1.

    OpenAIRE

    Diccianni, M B; Imagawa, M; Muramatsu, M

    1992-01-01

    Glutathione Transferase P (GST-P) gene expression is dominantly regulated by an upstream enhancer (GPEI) consisting of a dyad of palindromically oriented imperfect TPA (12-O-tetradecanoyl-phorbol-13-acetate)-responsive elements (TRE). GPEI is active in AP1-lacking F9 cells as well in AP1-containing HeLa cells. Despite GPEI's similarity to a TRE, c-jun co-transfection has only a minimal effect on transactivation. Antisense c-jun and c-fos co-transfection experiments further demonstrate the lac...

  8. Resistance to the Peptidyl Transferase Inhibitor Tiamulin Caused by Mutation of Ribosomal Protein L3

    OpenAIRE

    Bøsling, Jacob; Poulsen, Susan M.; Vester, Birte; Long, Katherine S.

    2003-01-01

    The antibiotic tiamulin targets the 50S subunit of the bacterial ribosome and interacts at the peptidyl transferase center. Tiamulin-resistant Escherichia coli mutants were isolated in order to elucidate mechanisms of resistance to the drug. No mutations in the rRNA were selected as resistance determinants using a strain expressing only a plasmid-encoded rRNA operon. Selection in a strain with all seven chromosomal rRNA operons yielded a mutant with an A445G mutation in the gene coding for ri...

  9. Generation of Active Bovine Terminal Deoxynucleotidyl Transferase (TdT in E.coli

    Directory of Open Access Journals (Sweden)

    Wee Liang Kuan

    2010-08-01

    Full Text Available A synthetic gene encoding bovine terminal deoxynucleotidyl transferase (TdT was generated, cloned into an expression vector and expressed in E.coli. The effects of altering culture and induction conditions on the nature of recombinant protein production were investigated. This led to the expression of active recombinant bovine TdT in E.coli. After purification and characterisation, the activity of the enzyme was assessed in a biological assay for apoptosis. The process described in this report enables the economical production of TdT for high throughput applications.

  10. Generation of Active Bovine Terminal Deoxynucleotidyl Transferase (TdT in E.coli

    Directory of Open Access Journals (Sweden)

    Wee Liang Kuan

    2010-01-01

    Full Text Available A synthetic gene encoding bovine terminal deoxynucleotidyl transferase (TdT was generated, cloned into an expression vector and expressed in E.coli. The effects of altering culture and induction conditions on the nature of recombinant protein production were investigated. This led to the expression of active recombinant bovine TdT in E.coli. After purification and characterisation, the activity of the enzyme was assessed in a biological assay for apoptosis. The process described in this report enables the economical production of TdT for high throughput applications.

  11. The O-GlcNAc Transferase Intellectual Disability Mutation L254F Distorts the TPR Helix.

    Science.gov (United States)

    Gundogdu, Mehmet; Llabrés, Salomé; Gorelik, Andrii; Ferenbach, Andrew T; Zachariae, Ulrich; van Aalten, Daan M F

    2018-05-17

    O-linked β-N-acetyl- D -glucosamine (O-GlcNAc) transferase (OGT) regulates protein O-GlcNAcylation, an essential post-translational modification that is abundant in the brain. Recently, OGT mutations have been associated with intellectual disability, although it is not understood how they affect OGT structure and function. Using a multi-disciplinary approach we show that the L254F OGT mutation leads to conformational changes of the tetratricopeptide repeats and reduced activity, revealing the molecular mechanisms contributing to pathogenesis. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

  12. Use of some inorganic and organic compounds as decontaminants for cobalt-60 and caesium-134 by clover plant grown on Inshas sandy soil

    Energy Technology Data Exchange (ETDEWEB)

    Abdel Sabour, M.F.; El-Naggar, H.A.; Soliman, S.M. (Atomic Energy Establishment, Cairo (Egypt). Nuclear Chemistry Dept.)

    1991-01-01

    Outdoor lysimeter experiments were carried out to elucidate the effect of 4 inorganic and 3 organic salts on {sup 60}Co and {sup 134}Cs uptake and dry matter yield of three cuts of clover in the soils of Inshas. A new concept was proposed for assessing the difference in capacity of soil to supply radionuclides to plants as affected by applied inorganic or organic compounds. A relation of plant tissue radionuclide content with time can be expressed in an exponential equation with corresponding regression coefficients. When the same plant species is grown on the same soil differing only in the applied compounds (e.g. Fe-EDDHA, Fe-DTPA, Fe-OAC, Fe(COO){sub 2}, Al(OH){sub 3}, Ca(H{sub 2}PO{sub 4}){sub 2}, Fe{sub 2}O{sub 3}) the derived equation will change. Differences of either Co or Cs accumulation with time between control and any other treatment were evaluated. The data also show that Fe-DTPA is preferred for {sup 60}Co as enhancing compound for plant uptake (for decontamination use), compared with other applied salts, on the other hand, any tested salts did not significantly effect {sup 134}Cs taken by clover. It was also found that more than 70% of the total cobalt uptake was accumulated in the roots which indicate that Co is less mobile in plants than Cs. (orig.).

  13. Use of some inorganic and organic compounds as decontaminants for cobalt-60 and caesium-134 by clover plant grown on Inshas sandy soil

    International Nuclear Information System (INIS)

    Abdel Sabour, M.F.; El-Naggar, H.A.; Soliman, S.M.

    1991-01-01

    Outdoor lysimeter experiments were carried out to elucidate the effect of 4 inorganic and 3 organic salts on 60 Co and 134 Cs uptake and dry matter yield of three cuts of clover in the soils of Inshas. A new concept was proposed for assessing the difference in capacity of soil to supply radionuclides to plants as affected by applied inorganic or organic compounds. A relation of plant tissue radionuclide content with time can be expressed in an exponential equation with corresponding regression coefficients. When the same plant species is grown on the same soil differing only in the applied compounds (e.g. Fe-EDDHA, Fe-DTPA, Fe-OAC, Fe(COO) 2 , Al(OH) 3 , Ca(H 2 PO 4 ) 2 , Fe 2 O 3 ) the derived equation will change. Differences of either Co or Cs accumulation with time between control and any other treatment were evaluated. The data also show that Fe-DTPA is preferred for 60 Co as enhancing compound for plant uptake (for decontamination use), compared with other applied salts, on the other hand, any tested salts did not significantly effect 134 Cs taken by clover. It was also found that more than 70% of the total cobalt uptake was accumulated in the roots which indicate that Co is less mobile in plants than Cs. (orig.) [de

  14. Resistance of red clover (Trifolium pratense) to the root parasitic plant Orobanche minor is activated by salicylate but not by jasmonate.

    Science.gov (United States)

    Kusumoto, Dai; Goldwasser, Yaakov; Xie, Xiaonan; Yoneyama, Kaori; Takeuchi, Yasutomo; Yoneyama, Koichi

    2007-09-01

    Obligate root holoparasites of the genus Orobanche attack dicotyledonous crops and cause severe losses in many parts of the world. Chemical induction of plant defence systems such as systemic acquired resistance was proposed to be an available strategy to control the root parasite, but the detailed mechanisms involved have not been clarified. The aim of this study was to elucidate the effects of salicylic acid (SA), jasmonic acid (JA) and their analogues on resistance of red clover to Orobanche parasitism. Roots of red clover grown in plastic chambers were applied with SA, S-methyl benzo[1,2,3]thiadiazole-7-carbothioate (BTH), methyl jasmonate (MeJA) and n-propyl dihydrojasmonate (PDJ), and then were inoculated with O. minor seeds. Attachments of the parasite were observed after 5 weeks. SA and BTH, inducers of SA-mediated defences, significantly reduced the number of established parasites by more than 75 %. By contrast, MeJA and PDJ, inducers of JA-mediated defences, did not affect parasitism. The reduction in the number of established parasites by SA and BTH was due to the inhibited elongation of O. minor radicles and the activation of defence responses in the host root including lignification of the endodermis. These results suggest that SA-induced resistance, but not JA-induced resistance, is effective in inhibiting Orobanche parasitism and that the resistance is expressed by the host root both externally and internally.

  15. Characterisation of the Candida albicans Phosphopantetheinyl Transferase Ppt2 as a Potential Antifungal Drug Target.

    Directory of Open Access Journals (Sweden)

    Katharine S Dobb

    Full Text Available Antifungal drugs acting via new mechanisms of action are urgently needed to combat the increasing numbers of severe fungal infections caused by pathogens such as Candida albicans. The phosphopantetheinyl transferase of Aspergillus fumigatus, encoded by the essential gene pptB, has previously been identified as a potential antifungal target. This study investigated the function of its orthologue in C. albicans, PPT2/C1_09480W by placing one allele under the control of the regulatable MET3 promoter, and deleting the remaining allele. The phenotypes of this conditional null mutant showed that, as in A. fumigatus, the gene PPT2 is essential for growth in C. albicans, thus fulfilling one aspect of an efficient antifungal target. The catalytic activity of Ppt2 as a phosphopantetheinyl transferase and the acyl carrier protein Acp1 as a substrate were demonstrated in a fluorescence transfer assay, using recombinant Ppt2 and Acp1 produced and purified from E.coli. A fluorescence polarisation assay amenable to high-throughput screening was also developed. Therefore we have identified Ppt2 as a broad-spectrum novel antifungal target and developed tools to identify inhibitors as potentially new antifungal compounds.

  16. Resistance to the peptidyl transferase inhibitor tiamulin caused by mutation of ribosomal protein l3.

    Science.gov (United States)

    Bøsling, Jacob; Poulsen, Susan M; Vester, Birte; Long, Katherine S

    2003-09-01

    The antibiotic tiamulin targets the 50S subunit of the bacterial ribosome and interacts at the peptidyl transferase center. Tiamulin-resistant Escherichia coli mutants were isolated in order to elucidate mechanisms of resistance to the drug. No mutations in the rRNA were selected as resistance determinants using a strain expressing only a plasmid-encoded rRNA operon. Selection in a strain with all seven chromosomal rRNA operons yielded a mutant with an A445G mutation in the gene coding for ribosomal protein L3, resulting in an Asn149Asp alteration. Complementation experiments and sequencing of transductants demonstrate that the mutation is responsible for the resistance phenotype. Chemical footprinting experiments show a reduced binding of tiamulin to mutant ribosomes. It is inferred that the L3 mutation, which points into the peptidyl transferase cleft, causes tiamulin resistance by alteration of the drug-binding site. This is the first report of a mechanism of resistance to tiamulin unveiled in molecular detail.

  17. Madumycin II inhibits peptide bond formation by forcing the peptidyl transferase center into an inactive state

    Energy Technology Data Exchange (ETDEWEB)

    Osterman, Ilya A.; Khabibullina, Nelli F.; Komarova, Ekaterina S.; Kasatsky, Pavel; Kartsev, Victor G.; Bogdanov, Alexey A.; Dontsova, Olga A.; Konevega, Andrey L.; Sergiev, Petr V.; Polikanov, Yury S. (InterBioScreen); (UIC); (MSU-Russia); (Kurchatov)

    2017-05-13

    The emergence of multi-drug resistant bacteria is limiting the effectiveness of commonly used antibiotics, which spurs a renewed interest in revisiting older and poorly studied drugs. Streptogramins A is a class of protein synthesis inhibitors that target the peptidyl transferase center (PTC) on the large subunit of the ribosome. In this work, we have revealed the mode of action of the PTC inhibitor madumycin II, an alanine-containing streptogramin A antibiotic, in the context of a functional 70S ribosome containing tRNA substrates. Madumycin II inhibits the ribosome prior to the first cycle of peptide bond formation. It allows binding of the tRNAs to the ribosomal A and P sites, but prevents correct positioning of their CCA-ends into the PTC thus making peptide bond formation impossible. We also revealed a previously unseen drug-induced rearrangement of nucleotides U2506 and U2585 of the 23S rRNA resulting in the formation of the U2506•G2583 wobble pair that was attributed to a catalytically inactive state of the PTC. The structural and biochemical data reported here expand our knowledge on the fundamental mechanisms by which peptidyl transferase inhibitors modulate the catalytic activity of the ribosome.

  18. Glutathione Transferases Superfamily: Cold-Inducible Expression of Distinct GST Genes in Brassica oleracea

    Directory of Open Access Journals (Sweden)

    Harshavardhanan Vijayakumar

    2016-07-01

    Full Text Available Plants, as sessile organisms, can suffer serious growth and developmental consequences under cold stress conditions. Glutathione transferases (GSTs, EC 2.5.1.18 are ubiquitous and multifunctional conjugating proteins, which play a major role in stress responses by preventing oxidative damage by reactive oxygen species (ROS. Currently, understanding of their function(s during different biochemical and signaling pathways under cold stress condition remain unclear. In this study, using combined computational strategy, we identified 65 Brassica oleracea glutathione transferases (BoGST and characterized them based on evolutionary analysis into 11 classes. Inter-species and intra-species duplication was evident between BoGSTs and Arabidopsis GSTs. Based on localization analyses, we propose possible pathways in which GST genes are involved during cold stress. Further, expression analysis of the predicted putative functions for GST genes were investigated in two cold contrasting genotypes (cold tolerance and susceptible under cold condition, most of these genes were highly expressed at 6 h and 1 h in the cold tolerant (CT and cold susceptible (CS lines, respectively. Overall, BoGSTU19, BoGSTU24, BoGSTF10 are candidate genes highly expressed in B. oleracea. Further investigation of GST superfamily in B. oleracea will aid in understanding complex mechanism underlying cold tolerance in plants.

  19. The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function

    International Nuclear Information System (INIS)

    Achilonu, Ikechukwu; Gildenhuys, Samantha; Fisher, Loren; Burke, Jonathan; Fanucchi, Sylvia; Sewell, B. Trevor; Fernandes, Manuel; Dirr, Heini W.

    2010-01-01

    The role of a topologically conserved isoleucine in the structure of glutathione transferase was investigated by replacing the Ile71 residue in human GSTA1-1 by alanine or valine. The common fold shared by members of the glutathione-transferase (GST) family has a topologically conserved isoleucine residue at the N-terminus of helix 3 which is involved in the packing of helix 3 against two β-strands in domain 1. The role of the isoleucine residue in the structure, function and stability of GST was investigated by replacing the Ile71 residue in human GSTA1-1 by alanine or valine. The X-ray structures of the I71A and I71V mutants resolved at 1.75 and 2.51 Å, respectively, revealed that the mutations do not alter the overall structure of the protein compared with the wild type. Urea-induced equilibrium unfolding studies using circular dichroism and tryptophan fluorescence suggest that the mutation of Ile71 to alanine or valine reduces the stability of the protein. A functional assay with 1-chloro-2,4-dinitrobenzene shows that the mutation does not significantly alter the function of the protein relative to the wild type. Overall, the results suggest that conservation of the topologically conserved Ile71 maintains the structural stability of the protein but does not play a significant role in catalysis and substrate binding

  20. Changes in amino transferases and muscle proteins when treating pigmeat with ionising rays

    Energy Technology Data Exchange (ETDEWEB)

    Hamm, R; Hofmann, K; Gruenewald, T; Partmann, W [Bundesanstalt fuer Fleischforschung, Kulmbach (F.R. Germany). Inst. fuer Chemie und Physik; Bundesforschungsanstalt fuer Ernaehrung, Karlsruhe [F.R. Germany

    1975-01-01

    Slices of lean pigmeat were treated with electron beams doses of 0.2, 1.0 and 5.0 Mrad. Low irradiation doses led to an increase in the activity of aspartate amino transferase (GOT) and alanin amino transferase (GPT) in the tissue generally and in the sarcoplasm (juice expressed from the muscle). 5 Mrad caused a great reduction in the activity of GOT and GPT in the tissue and the sarcoplasm. It seems doubtful whether this inactivation is due to a destruction of enzyme sulphhydryl groups. Irradiating with 5 Mrad resulted in partial release of the mitochondrial GOT isozyme (GOTsub(M)) into the sarcoplasm. This indicates damage to the mitochondrial membranes by ionising radiation. Irradiating the pigmeat increased the pH of the tissue and lowered its water binding ability (increase in drip). Up to a dose of 1 Mrad the solubility of the sarcoplasmic proteins was not definitely affected, but 5 Mrad caused a considerable drop in protein solubility. Surprisingly a dose of even 5 Mrad did not change the total number of sulphhydryl groups present in the tissue. Sephadex thin layer electrophoresis showed that at 0.2 Mrad there was a drastic decrease in the myosin band and an increase in peptide fragments of low molecular weight, whilst actin was little changed.

  1. Changes in amino transferases and muscle proteins when treating pigmeat with ionising rays

    International Nuclear Information System (INIS)

    Hamm, R.; Hofmann, K.; Gruenewald, T.; Partmann, W.

    1975-01-01

    Slices of lean pigmeat were treated with electron beams doses of 0.2, 1.0 and 5.0 Mrad. Low irradiation doses led to an increase in the activity of aspartate amino transferase (GOT) and alanin amino transferase (GPT) in the tissue generally and in the sarcoplasm (juice expressed from the muscle). 5 Mrad caused a great reduction in the activity of GOT and GPT in the tissue and the sarcoplasm. It seems doubtful whether this inactivation is due to a destruction of enzyme sulphhydryl groups. Irradiating with 5 Mrad resulted in partial release of the mitochondrial GOT isozyme (GOTsub(M)) into the sarcoplasm. This indicates damage to the mitochondrial membranes by ionising radiation. Irradiating the pigmeat increased the pH of the tissue and lowered its water binding ability (increase in drip). Up to a dose of 1 Mrad the solubility of the sarcoplasmic proteins was not definitely affected, but 5 Mrad caused a considerable drop in protein solubility. Surprisingly a dose of even 5 Mrad did not change the total number of sulphhydryl groups present in the tissue. Sephadex thin layer electrophoresis showed that at 0.2 Mrad there was a drastic decrease in the myosin band and an increase in peptide fragments of low molecular weight, whilst actin was little changed. (orig.) [de

  2. Epidermal growth factor regulation of glutathione S-transferase gene expression in the rat is mediated by class Pi glutathione S-transferase enhancer I.

    Science.gov (United States)

    Matsumoto, M; Imagawa, M; Aoki, Y

    2000-07-01

    Using chloramphenicol acetyltransferase assays we showed that epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha), and 3,3',4,4',5-pentachlorobiphenyl (PenCB) induce class Pi glutathione S-transferase (GSTP1) in primary cultured rat liver parenchymal cells. GSTP1 enhancer I (GPEI), which is required for the stimulation of GSTP1 expression by PenCB, also mediates EGF and TGF alpha stimulation of GSTP1 gene expression. However, hepatocyte growth factor and insulin did not stimulate GPEI-mediated gene expression. On the other hand, the antioxidant reagents butylhydroxyanisole and t-butylhydroquinone, stimulated GPEI-mediated gene expression, but the level of GSTP1 mRNA was not elevated. Our observations suggest that EGF and TGF alpha induce GSTP1 by the same signal transduction pathway as PenCB. Since the sequence of GPEI is similar to that of the antioxidant responsive element (ARE), some factors which bind to ARE might play a role in GPEI-mediated gene expression.

  3. Immunogold Labelling to Localize Polyphenol Oxidase (PPO) During Wilting of Red Clover Leaf Tissue and the Effect of Removing Cellular Matrices on PPO Protection of Glycerol-Based Lipid in the Rumen

    Science.gov (United States)

    The enzyme polyphenol oxidase (PPO) reduces the extent of proteolysis and lipolysis within red clover fed to ruminants. PPO catalyses the conversion of phenols to quinones which can react with nucleophilic cellular constituents (e.g. proteins), forming protein-phenol complexes that may reduce protei...

  4. Changes in the spore numbers of AM fungi and in AM colonisation of roots of clovers and grasses on a peat-muck soil with respect to mineral fertilization

    International Nuclear Information System (INIS)

    Kowalska, T. K.; Kwiatkowaska, E.

    2016-01-01

    A 4-year plot experiment was conducted to determine the dynamics of changes in the spore density of arbuscular mycorrhizal fungi (AMF) and of the degree of endomycorrhizal colonisation of roots of clovers and meadow grasses on an organic peat-muck soil in a post-marshy habitat, taking into account the effect of mineral fertilisation (NPK). The experimental object comprised four plots that represented the fertilisation treatments, sown with white clover (Trifolium repens L.), red clover (Trifolium pratense L.), smooth meadow-grass (Poa pratensis L.), and a mix of grasses composed of perennial ryegrass (Lolium perenne L.), meadow fescue (Festuca pratensis Huds.), smooth meadow-grass (Poa pratensis L.), and cocksfoot (Dactylis glomerata L.). Analogous sowing was performed on control (non-fertilised) plots. It was found that spores of AMF occurred in 100 percent of the samples of the soil studied, and the average total number of AMF spores isolated from soil under the particular plant combinations was high and amounted to 1858 spores (range from 1392 to 2443) in 100 g of air-dried soil. The percentage share of the clover and grass roots colonised by indigenous endomycorrhizal fungi was very low and varied from 0 to 46 (average from 4.1 percent to 12.2 percent). No correlation was found between the spore numbers of AMF in the soil and the degree of mycorrhized roots of the clovers and grasses. Mineral fertilisation stimulated the sporulation of AM fungi but had no effect on root colonisation by these fungi. (author)

  5. Reviewing Hit Discovery Literature for Difficult Targets: Glutathione Transferase Omega-1 as an Example.

    Science.gov (United States)

    Xie, Yiyue; Dahlin, Jayme L; Oakley, Aaron J; Casarotto, Marco G; Board, Philip G; Baell, Jonathan B

    2018-05-10

    Early stage drug discovery reporting on relatively new or difficult targets is often associated with insufficient hit triage. Literature reviews of such targets seldom delve into the detail required to critically analyze the associated screening hits reported. Here we take the enzyme glutathione transferase omega-1 (GSTO1-1) as an example of a relatively difficult target and review the associated literature involving small-molecule inhibitors. As part of this process we deliberately pay closer-than-usual attention to assay interference and hit quality aspects. We believe this Perspective will be a useful guide for future development of GSTO1-1 inhibitors, as well serving as a template for future review formats of new or difficult targets.

  6. Response of Glutathione and Glutathione S-transferase in Rice Seedlings Exposed to Cadmium Stress

    Directory of Open Access Journals (Sweden)

    Chun-hua ZHANG

    2008-03-01

    Full Text Available A hydroponic culture experiment was done to investigate the effect of Cd stress on glutathione content (GSH and glutathione S-transferase (GST, EC 2.5.1.18 activity in rice seedlings. The rice growth was severely inhibited when Cd level in the solution was higher than 10 mg/L. In rice shoots, GSH content and GST activity increased with the increasing Cd level, while in roots, GST was obviously inhibited by Cd treatments. Compared with shoots, the rice roots had higher GSH content and GST activity, indicating the ability of Cd detoxification was much higher in roots than in shoots. There was a significant correlation between Cd level and GSH content or GST activity, suggesting that both parameters may be used as biomarkers of Cd stress in rice.

  7. Differential roles of tau class glutathione S-transferases in oxidative stress

    DEFF Research Database (Denmark)

    Kilili, Kimiti G; Atanassova, Neli; Vardanyan, Alla

    2004-01-01

    The plant glutathione S-transferase BI-GST has been identified as a potent inhibitor of Bax lethality in yeast, a phenotype associated with oxidative stress and disruption of mitochondrial functions. Screening of a tomato two-hybrid library for BI-GST interacting proteins identified five homologous...... Tau class GSTs, which readily form heterodimers between them and BI-GST. All six LeGSTUs were found to be able to protect yeast cells from prooxidant-induced cell death. The efficiency of each LeGSTU was prooxidant-specific, indicating a different role for each LeGSTU in the oxidative stress......-response mechanism. The prooxidant protective effect of all six proteins was suppressed in the absence of YAP1, a transcription factor that regulates hydroperoxide homeostasis in Saccharomyces cerevisiae, suggesting a role for the LeGSTUs in the context of the YAP1-dependent stress-responsive machinery...

  8. Phi Class of Glutathione S-transferase Gene Superfamily Widely Exists in Nonplant Taxonomic Groups.

    Science.gov (United States)

    Munyampundu, Jean-Pierre; Xu, You-Ping; Cai, Xin-Zhong

    2016-01-01

    Glutathione S-transferases (GSTs) constitute a superfamily of enzymes involved in detoxification of noxious compounds and protection against oxidative damage. GST class Phi (GSTF), one of the important classes of plant GSTs, has long been considered as plant specific but was recently found in basidiomycete fungi. However, the range of nonplant taxonomic groups containing GSTFs remains unknown. In this study, the distribution and phylogenetic relationships of nonplant GSTFs were investigated. We identified GSTFs in ascomycete fungi, myxobacteria, and protists Naegleria gruberi and Aureococcus anophagefferens. GSTF occurrence in these bacteria and protists correlated with their genome sizes and habitats. While this link was missing across ascomycetes, the distribution and abundance of GSTFs among ascomycete genomes could be associated with their lifestyles to some extent. Sequence comparison, gene structure, and phylogenetic analyses indicated divergence among nonplant GSTFs, suggesting polyphyletic origins during evolution. Furthermore, in silico prediction of functional partners suggested functional diversification among nonplant GSTFs.

  9. Echinococcus granulosus: Evidence of a heterodimeric glutathione transferase built up by phylogenetically distant subunits.

    Science.gov (United States)

    Arbildi, Paula; La-Rocca, Silvana; Lopez, Veronica; Da-Costa, Natalia; Fernandez, Veronica

    2017-01-01

    In the cestode parasite Echinococcus granulosus, three phylogenetically distant cytosolic glutathione transferases (GSTs) (EgGST1, 2 and 3) were identified. Interestingly, the C-terminal domains of EgGST3 and EgGST2 but not EgGST1, exhibit all amino acids involved in Sigma-class GST dimerization. Here, we provide evidence indicating that EgGST2 and EgGST3 naturally form a heterodimeric structure (EgGST2-3), and also we report the enzymatic activity of the recombinant heterodimer. EgGST2-3 might display novel properties able to influence the infection establishment. This is the first report of a stable heterodimeric GST built up by phylogenetically distant subunits. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Isolation and purification of glutathione S-transferases from Brachionus plicatilis and B. calyciflorus (Rotifera).

    Science.gov (United States)

    Bowman, B P; Snell, T W; Cochrane, B J

    1990-01-01

    1. The enzyme glutathione S-transferase (GST), a critical element in xenobiotic metabolism, was isolated from the marine rotifer Brachionus plicatilis and its freshwater congener B. calyciflorus. 2. In B. plicatilis, GST comprised 4.2% of cytosolic protein and was present as three separate isozymes with mol. wts 30,000, 31,400 and 33,700. Specific activity of crude homogenates was 56 nmol min-1 mg-1 protein, while that of affinity chromatography purified GST was 1850. 3. In B. calyciflorus, GST was present as two isozymes with mol. wts of 26,300 and 28,500, representing 1.0% of cytosolic protein. Crude GST specific activity was 1750 nmol min-1 mg-1 protein and purified was 72,400. 4. Rotifer GSTs are unusual because they are monomers whereas all other animals thus far investigated posses dimeric GSTs.

  11. Nuclear translocation of glutathione transferase omega is a progression marker in Barrett's esophagus

    DEFF Research Database (Denmark)

    Piaggi, Simona; Marchi, Santino; Ciancia, Eugenio

    2009-01-01

    Barrett's esophagus (BE) represents a major risk factor for esophageal adenocarcinoma (AC). For this reason, patients with BE are subjected to a systematic endoscopic surveillance to detect initial evolution towards non-invasive neoplasia (NiN) and cancer, that eventually occurs only in a small......-S-transferase-omega 1 could be involved in the stress response of human cells playing a role in the cancer progression of Barrett's esophagus. Its immunohistochemical detection could represent a useful tool in the grading of Barrett's disease.......N in BE and to understand the mechanisms of the progression from BE to AC. We investigated the expression and subcellular localization of GSTO1 in biopsies from patients with BE and in human cancer cell lines subjected to heath shock treatment. A selective nuclear localisation of GSTO1 was found in 16/16 biopsies with low...

  12. Glutathione transferases are structural and functional outliers in the thioredoxin fold.

    Science.gov (United States)

    Atkinson, Holly J; Babbitt, Patricia C

    2009-11-24

    Glutathione transferases (GSTs) are ubiquitous scavengers of toxic compounds that fall, structurally and functionally, within the thioredoxin fold suprafamily. The fundamental catalytic capability of GSTs is catalysis of the nucleophilic addition or substitution of glutathione at electrophilic centers in a wide range of small electrophilic compounds. While specific GSTs have been studied in detail, little else is known about the structural and functional relationships between different groupings of GSTs. Through a global analysis of sequence and structural similarity, it was determined that variation in the binding of glutathione between the two major subgroups of cytosolic (soluble) GSTs results in a different mode of glutathione activation. Additionally, the convergent features of glutathione binding between cytosolic GSTs and mitochondrial GST kappa are described. The identification of these structural and functional themes helps to illuminate some of the fundamental contributions of the thioredoxin fold to catalysis in the GSTs and clarify how the thioredoxin fold can be modified to enable new functions.

  13. Glutathione-binding site of a bombyx mori theta-class glutathione transferase.

    Directory of Open Access Journals (Sweden)

    M D Tofazzal Hossain

    Full Text Available The glutathione transferase (GST superfamily plays key roles in the detoxification of various xenobiotics. Here, we report the isolation and characterization of a silkworm protein belonging to a previously reported theta-class GST family. The enzyme (bmGSTT catalyzes the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, 1,2-epoxy-3-(4-nitrophenoxy-propane, and 4-nitrophenethyl bromide. Mutagenesis of highly conserved residues in the catalytic site revealed that Glu66 and Ser67 are important for enzymatic function. These results provide insights into the catalysis of glutathione conjugation in silkworm by bmGSTT and into the metabolism of exogenous chemical agents.

  14. Expression of polypeptide GalNAc-transferases in stratified epithelia and squamous cell carcinomas

    DEFF Research Database (Denmark)

    Mandel, U; Hassan, H; Therkildsen, M H

    1999-01-01

    GalNAc-T1, -T2, and -T3. Application of this panel of novel antibodies revealed that GalNAc- transferases are differentially expressed in different cell lines, in spermatozoa, and in oral mucosa and carcinomas. For example, GalNAc-T1 and -T2 but not -T3 were highly expressed in WI38 cells, and Gal......NAc-T3 but not GalNAc-T1 or -T2 was expressed in spermatozoa. The expression patterns in normal oral mucosa were found to vary with cell differentiation, and for GalNAc-T2 and -T3 this was reflected in oral squamous cell carcinomas. The expression pattern of GalNAc-T1 was on the other hand changed...

  15. Identification of Protein-Protein Interactions with Glutathione-S-Transferase (GST) Fusion Proteins.

    Science.gov (United States)

    Einarson, Margret B; Pugacheva, Elena N; Orlinick, Jason R

    2007-08-01

    INTRODUCTIONGlutathione-S-transferase (GST) fusion proteins have had a wide range of applications since their introduction as tools for synthesis of recombinant proteins in bacteria. GST was originally selected as a fusion moiety because of several desirable properties. First and foremost, when expressed in bacteria alone, or as a fusion, GST is not sequestered in inclusion bodies (in contrast to previous fusion protein systems). Second, GST can be affinity-purified without denaturation because it binds to immobilized glutathione, which provides the basis for simple purification. Consequently, GST fusion proteins are routinely used for antibody generation and purification, protein-protein interaction studies, and biochemical analysis. This article describes the use of GST fusion proteins as probes for the identification of protein-protein interactions.

  16. Serum glutathione transferase does not respond to indole-3-carbinol: A pilot study

    Directory of Open Access Journals (Sweden)

    Daniel R McGrath

    2010-05-01

    Full Text Available Daniel R McGrath1, Hamid Frydoonfar2, Joshua J Hunt3, Chris J Dunkley3, Allan D Spigelman41Ipswich Hospital, Ipswich, UK; 2Hunter Pathology Service, New South Wales; 3Royal Newcastle Centre, Newcastle; 4St Vincent’s Clinical School, Sydney, AustraliaBackground: Despite the well recognized protective effect of cruciferous vegetables against various cancers, including human colorectal cancers, little is known about how this effect is conferred. It is thought that some phytochemicals found only in these vegetables confer the protection. These compounds include the glucosinolates, of which indole-3-carbinol is one. They are known to induce carcinogen-metabolizing (phase II enzymes, including the glutathione S-transferase (GST family. Other effects in humans are not well documented. We wished to assess the effect of indole-3-carbinol on GST enzymes.Methods: We carried out a placebo-controlled human volunteer study. All patients were given 400 mg daily of indole-3-carbinol for three months, followed by placebo. Serum samples were tested for the GSTM1 genotype by polymerase chain reaction. Serum GST levels were assessed using enzyme-linked immunosorbent assay and Western Blot methodologies.Results: Forty-nine volunteers completed the study. GSTM1 genotypes were obtained for all but two volunteers. A slightly greater proportion of volunteers were GSTM1-positive, in keeping with the general population. GST was detected in all patients. Total GST level was not affected by indole-3-carbinol dosing compared with placebo. Although not statistically significant, the GSTM1 genotype affected the serum GST level response to indole-3-carbinol.Conclusion: Indole-3-carbinol does not alter total serum GST levels during prolonged dosing.Keywords: pilot study, colorectal cancer, glutathione transferase, human, indole-3-carbinol

  17. Loci Affecting Gamma-Glutamyl Transferase in Adults and Adolescents Show Age X SNP interaction and Cardiometabolic Disease Associations

    NARCIS (Netherlands)

    Middelberg, R.P.S.; Benyamin, B.; de Moor, M.H.M.; Warrington, N.M.; Gordon, S.; Henders, A.K.; Medland, S.E.; Nyholt, DR; de Geus, E.J.C.; Hottenga, J.J.; Willemsen, G.; Beilin, L.J.; Mori, T.A.; Wright, M.J.; Heath, A.C.; Madden, P.A.F.; Boomsma, D.I.; Pennell, C.E.; Montgomery, G.W.; Martin, N.G.; Whitfield, J.B.

    2012-01-01

    Serum gamma-glutamyl transferase (GGT) activity is a marker of liver disease which is also prospectively associated with the risk of all-cause mortality, cardiovascular disease, type 2 diabetes and cancers. We have discovered novel loci affecting GGT in a genome-wide association study (rs1497406 in

  18. Systemic catechol-O-methyl transferase inhibition enables the D1 agonist radiotracer R-[11C]SKF 82957

    DEFF Research Database (Denmark)

    Palner, Mikael; McCormick, Patrick; Parkes, Jun

    2010-01-01

    R-[(11)C]-SKF 82957 is a high-affinity and potent dopamine D(1) receptor agonist radioligand, which gives rise to a brain-penetrant lipophilic metabolite. In this study, we demonstrate that systemic administration of catechol-O-methyl transferase (COMT) inhibitors blocks this metabolic pathway, f...

  19. Glutathione S-transferase M1 and T1, CYP1A2-2467T/delT ...

    African Journals Online (AJOL)

    The present study investigated the impact of metabolic gene polymorphisms in modulating lung cancer risk susceptibility. Gene polymorphisms encoding Cytochrome 1A2 (CYP1A2) and Glutathione-S-transferases (GSTT1 and GSTM1) are involved in the bioactivation and detoxification of tobacco carcinogens and may ...

  20. Mapping important nucleotides in the peptidyl transferase centre of 23 S rRNA using a random mutagenesis approach

    DEFF Research Database (Denmark)

    Porse, B T; Garrett, R A

    1995-01-01

    Random mutations were generated in the lower half of the peptidyl transferase loop in domain V of 23 S rRNA from Escherichia coli using a polymerase chain reaction (PCR) approach, a rapid procedure for identifying mutants and a plasmid-based expression system. The effects of 21 single-site mutati...

  1. Cloning and characterization of a glutathione S-transferase homologue from the plant pathogenic fungus Botrytis cinerea

    NARCIS (Netherlands)

    Prins, T.W.; Wagemakers, L.; Schouten, A.; Kan, van J.A.L.

    2000-01-01

    A gene was cloned from Botrytis cinerea that encodes a protein homologous to glutathione S-transferase (GST). The gene, denominated Bcgst1, is present in a single copy and represents the first example of such a gene from a filamentous fungus. The biochemical function of GSTs is to conjugate toxic

  2. The pleuromutilin drugs tiamulin and valnemulin bind to the RNA at the peptidyl transferase centre on the ribosome.

    Science.gov (United States)

    Poulsen, S M; Karlsson, M; Johansson, L B; Vester, B

    2001-09-01

    The pleuromutilin antibiotic derivatives, tiamulin and valnemulin, inhibit protein synthesis by binding to the 50S ribosomal subunit of bacteria. The action and binding site of tiamulin and valnemulin was further characterized on Escherichia coli ribosomes. It was revealed that these drugs are strong inhibitors of peptidyl transferase and interact with domain V of 23S RNA, giving clear chemical footprints at nucleotides A2058-9, U2506 and U2584-5. Most of these nucleotides are highly conserved phylogenetically and functionally important, and all of them are at or near the peptidyl transferase centre and have been associated with binding of several antibiotics. Competitive footprinting shows that tiamulin and valnemulin can bind concurrently with the macrolide erythromycin but compete with the macrolide carbomycin, which is a peptidyl transferase inhibitor. We infer from these and previous results that tiamulin and valnemulin interact with the rRNA in the peptidyl transferase slot on the ribosomes in which they prevent the correct positioning of the CCA-ends of tRNAs for peptide transfer.

  3. The interdomain flexible linker of the polypeptide GalNAc transferases dictates their long-range glycosylation preferences

    DEFF Research Database (Denmark)

    Rivas, Matilde De Las; Lira-Navarrete, Erandi; Daniel, Earnest James Paul

    2017-01-01

    The polypeptide GalNAc-transferases (GalNAc-Ts), that initiate mucin-type O-glycosylation, consist of a catalytic and a lectin domain connected by a flexible linker. In addition to recognizing polypeptide sequence, the GalNAc-Ts exhibit unique long-range N- A nd/or C-terminal prior glycosylation ...

  4. Transmutation of human glutathione transferase A2-2 with peroxidase activity into an efficient steroid isomerase.

    Science.gov (United States)

    Pettersson, Par L; Johansson, Ann-Sofie; Mannervik, Bengt

    2002-08-16

    A major goal in protein engineering is the tailor-making of enzymes for specified chemical reactions. Successful attempts have frequently been based on directed molecular evolution involving libraries of random mutants in which variants with desired properties were identified. For the engineering of enzymes with novel functions, it would be of great value if the necessary changes of the active site could be predicted and implemented. Such attempts based on the comparison of similar structures with different substrate selectivities have previously met with limited success. However, the present work shows that the knowledge-based redesign restricted to substrate-binding residues in human glutathione transferase A2-2 can introduce high steroid double-bond isomerase activity into the enzyme originally characterized by glutathione peroxidase activity. Both the catalytic center activity (k(cat)) and catalytic efficiency (k(cat)/K(m)) match the values of the naturally evolved glutathione transferase A3-3, the most active steroid isomerase known in human tissues. The substrate selectivity of the mutated glutathione transferase was changed 7000-fold by five point mutations. This example demonstrates the functional plasticity of the glutathione transferase scaffold as well as the potential of rational active-site directed mutagenesis as a complement to DNA shuffling and other stochastic methods for the redesign of proteins with novel functions.

  5. The association of alcohol intake with gamma-glutamyl transferase (GGT) levels: Evidence for correlated genetic effects

    NARCIS (Netherlands)

    van Beek, J.H.D.A.; de Moor, M.H.M.; Geels, L.M.; Sinke, M.R.T.; de Geus, E.J.C.; Lubke, G.H.; Kluft, C.; Neuteboom, J.; Vink, J.M.; Willemsen, G.; Boomsma, D.I.

    2014-01-01

    Background: Blood levels of gamma-glutamyl transferase (GGT) are used as a marker for (heavy) alcohol use. The role of GGT in the anti-oxidant defense mechanism that is part of normal metabolism supposes a causal effect of alcohol intake on GGT. However, there is variability in the response of GGT

  6. Cooperation of terminal deoxynucleotidyl transferase with DNA polymerase α in the replication of ultraviolet-irradiated DNA

    International Nuclear Information System (INIS)

    Yoshida, S.; Masaki, S.; Nakamura, H.; Morita, T.

    1981-01-01

    The amount of DNA synthesis in vitro with the ultraviolet-irradiated poly(dT).oligo(rA) template initiators catalysed by DNA polymerase α (Masaki, S. and Yoshida, S., Biochim. Biophys. Acta 521, 74-88) decreased with the dose of ultraviolet-irradiation. The ultraviolet irradiation to the template, however, did not affect the rate of incorporation of incorrect deoxynucleotides into the newly synthesized poly(dA). The addition of terminal deoxynucleotidyl transferase to this system enhanced the DNA synthesis to a level which is comparable to that of the control and it concomitantly increased the incorporation of the mismatched deoxynucleotide into the newly synthesized poly(dA) strands. On the other hand, with an unirradiated template initiator, the misincorporation was only slightly enhanced by the addition of terminal deoxynucleotidyl transferase. The sizes of newly synthesized DNA measured by the sedimentation velocities were found to be smaller with the ultraviolet-irradiated templates but they increased to the control level with the addition of terminal deoxynucleotidyl transferase to the systems. These results suggest that terminal deoxynucleotidyl transferase can help DNA polymerase α to bypass thymine dimers in vitro by the formation of mismatched regions at the positions opposite to pyrimidine dimers on the template. (Auth.)

  7. Mutations in N-acetylglucosamine (O-GlcNAc) transferase in patients with X-linked intellectual disability

    NARCIS (Netherlands)

    Willems, A.P.; Gundogdu, M.; Kempers, M.J.E.; Giltay, J.C.; Pfundt, R.P.; Elferink, M.; Loza, B.F.; Fuijkschot, J.; Ferenbach, A.T.; Gassen, K.L. van; Aalten, D.M.F. van; Lefeber, D.J.

    2017-01-01

    N-Acetylglucosamine (O-GlcNAc) transferase (OGT) regulates protein O-GlcNAcylation, an essential and dynamic post-translational modification. The O-GlcNAc modification is present on numerous nuclear and cytosolic proteins and has been implicated in essential cellular functions such as signaling and

  8. Function and phylogeny of bacterial butyryl-CoA:acetate transferases and their diversity in the proximal colon of swine

    Science.gov (United States)

    Studying the host-associated butyrate-producing bacterial community is important because butyrate is essential for colonic homeostasis and gut health. Previous research has identified the butyryl-coA:acetate transferase (2.3.8.3) as a the main gene for butyrate production in intestinal ecosystems; h...

  9. Nitrate leaching and residual effect in dairy crop rotations with grass-clover leys as influenced by sward age, grazing, cutting and fertilizer regimes

    DEFF Research Database (Denmark)

    Eriksen, Jørgen; Askegaard, Margrethe; Rasmussen, Jim

    2015-01-01

    Intensive dairy farming, with grass-arable crop rotations is challenged by low N use efficiency that may have adverse environmental consequences. We investigated nitrate leaching and N fertility effects of grass–clover leys for five years in two organic crop rotations with different grassland...... with the 2-yr-old leaching the most (36–46 kg N ha−1), and (4) high leaching (>50 kg N ha−1) with lupin and maize, where especially maize was consistently high in all five years (average 81 kg N ha−1). Great care should be taken during all phases of the dairy crop rotation where grasslands cause considerable...... build-up of fertility. With due care and the best management practice, nitrate leaching losses may be reduced to low levels....

  10. Environmental monitoring near the Macaulay Point and Clover Point marine sewage outfalls at Victoria, British Columbia in 1989 and 1990. Regional data report No. DR 92-14

    Energy Technology Data Exchange (ETDEWEB)

    Colodey, A G; Salmon, R A; Lim, P G

    1992-01-01

    The marine environment surrounding the Capital Region district (CRD) of British Columbia that includes the City of Victoria has been monitored by municipal, provincial, and federal authorities over the past 20 years to determine the impact of wastewater discharges. This report presents data from limited sampling conducted near the main CRD deep-water outfalls at Clover and Macaulay Points, the Canadian Department of National Defence septic tank outfall off Belmont and Esquimault Lagoon, and reference locations on Constance Bank and Discovery Island. Samples for this study were collected in April and October 1989, and April 1990. Parameters investigated were sediment and biota trace metals, sediment particle size, sediment volatile residues, visual descriptions of sediment, bacterial counts, water conductivity, temperature, salinity, and dissolved oxygen.

  11. Forage fiber effects on particle size reduction, ruminal stratification, and selective retention in heifers fed highly digestible grass/clover silages

    DEFF Research Database (Denmark)

    Schulze, Anne-Katrine Skovsted; Weisbjerg, Martin Riis; Storm, Adam Christian

    2014-01-01

    The objective of this study was to investigate the effect of NDF content in highly digestible grass/clover silage on particle size reduction, ruminal stratification, and selective retention in dairy heifers. The reduction in particle size from feed to feces was evaluated and related to feed intake...... measured. Intake of NDF increased linearly from 2.3 to 2.8 kg/d with greater NDF content of forages (P = 0.01), but silages were exposed to similar eating time (P = 0.55) and rumination time per kg NDF (P = 0.35). No linear effect of NDF content was found on proportion of LP in ingested feed boluses (P = 0.......31), medial rumen digesta (P = 0.95), ventral rumen digesta (P = 0.84), and feces (P = 0.09). Greater proportions of DM (P ruminal digesta compared with ventral rumen, and differences in DM proportion increased with greater NDF content (P = 0...

  12. Effects of maturity and harvest season of grass-clover silage and of forage-to-concentrate ratio on milk production of dairy cows

    DEFF Research Database (Denmark)

    Alstrup, L; Søegaard, K; Weisbjerg, M R

    2016-01-01

    This study examined the effects of maturity and season of harvest of grass-clover silages and forage:concentrate ratio (FCR) on feed intake, milk production, chewing activity, digestibility, and fecal consistency of Holstein dairy cows. Comparison included 2 cuts in spring season (early and late......) and 2 cuts in summer season (early and late) combined with high FCR (80:20; HFCR) and low FCR (50:50; LFCR). The experiment included 24 lactating Holstein cows arranged as 2 repeated 4 × 4 Latin squares with four 21-d periods and included measurements of feed composition, feed intake, milk production...... digestible than late maturity cuts, which was also reflected in a lower concentration of neutral detergent fiber (NDF) in early maturity cuts, whereas summer cuts had a higher crude protein concentration than spring cuts. Increased maturity decreased the intake of DM and energy, increased NDF intake...

  13. Use of inorganic and organic compounds as decontaminates for cobalt T-60 and cesium-134 by clover plant grown on Inshas Sandy soil

    International Nuclear Information System (INIS)

    Abdel-Sabour, M.F.; El-naggr, H.A.; Soliman, S.M.

    1994-01-01

    Out door lysimeter experiment was carried out to elucidate the effect of four inorganic and three organic salts on 60 Co and 134 Cs uptake and dry matter yield of three cuts of clover in the soil Inshas A new concept was proposed for assessing the difference in capacity of soil to supply radionuclides (i.e. 60 Co and 134 Cs) to plants as affected by applied inorganic or organic compounds. The relation of plant tissues (P c ) Co/IR Cs content with increasing time can be expressed as log (Pc) = A + B log time (T), where A and B the regression. When the same plant species is grown on the same soil differ only in the applied compound (e.g. Fe-EDDHA, Fe-DTPA, Fe-O A C, Fe(Coo)z, A1(OH)3, Ca(H2 P O 4 )2 and Fe 2 O 3 ) the equation will change to, log (P'c) = A + B' log (T) (I). Based on both equations, the relationship between (Pc) becomes; log (P'c) = [(A'B')/(A B + A B ) [ +[1/A'B + A B ) [ log (Pc) (II). The intercept (C) and slope (S) in Eq. (II) were determined among 7 treatments (for either 60 Co or 134 Cs). Then the difference of either Co or Cs accumulation with time between control and any other treatment was evaluated according to (C) and (S) values. The data also that Fe-DTPA is preferred for 60 Co as enhancing compound for plant uptake (for decontamination use); compared with other applied salts; on the other hand, any tested salts did not significantly effect 134 Cs taken by clover. Also, it was found that more than 70% of total cobalt uptake was accumulated in the roots which indicate that Co is less mobile in plants than Cs. 3 figs., 4 tabs

  14. High spatial variation in population size and symbiotic performance of Rhizobium leguminosarum bv. trifolii with white clover in New Zealand pasture soils

    Science.gov (United States)

    Tillard, Guyléne; van Ham, Robert; Ballard, Ross; Farquharson, Elizabeth; Gerard, Emily; Geurts, Rene; Brown, Matthew; Ridgway, Hayley; O’Callaghan, Maureen

    2018-01-01

    Biological nitrogen fixation through the legume-rhizobia symbiosis is important for sustainable pastoral production. In New Zealand, the most widespread and valuable symbiosis occurs between white clover (Trifolium repens L.) and Rhizobium leguminosarum bv. trifolii (Rlt). As variation in the population size (determined by most probable number assays; MPN) and effectiveness of N-fixation (symbiotic potential; SP) of Rlt in soils may affect white clover performance, the extent in variation in these properties was examined at three different spatial scales: (1) From 26 sites across New Zealand, (2) at farm-wide scale, and (3) within single fields. Overall, Rlt populations ranged from 95 to >1 x 108 per g soil, with variation similar at the three spatial scales assessed. For almost all samples, there was no relationship between rhizobia population size and ability of the population to fix N during legume symbiosis (SP). When compared with the commercial inoculant strain, the SP of soils ranged between 14 to 143% efficacy. The N-fixing ability of rhizobia populations varied more between samples collected from within a single hill country field (0.8 ha) than between 26 samples collected from diverse locations across New Zealand. Correlations between SP and calcium and aluminium content were found in all sites, except within a dairy farm field. Given the general lack of association between SP and MPN, and high spatial variability of SP at single field scale, provision of advice for treating legume seed with rhizobia based on field-average MPN counts needs to be carefully considered. PMID:29489845

  15. Knock-down of transcript abundance of a family of Kunitz proteinase inhibitor genes in white clover (Trifolium repens) reveals a redundancy and diversity of gene function.

    Science.gov (United States)

    Islam, Afsana; Leung, Susanna; Burgess, Elisabeth P J; Laing, William A; Richardson, Kim A; Hofmann, Rainer W; Dijkwel, Paul P; McManus, Michael T

    2015-12-01

    The transcriptional regulation of four phylogenetically distinct members of a family of Kunitz proteinase inhibitor (KPI) genes isolated from white clover (Trifolium repens; designated Tr-KPI1, Tr-KPI2, Tr-KPI4 and Tr-KPI5) has been investigated to determine their wider functional role. The four genes displayed differential transcription during seed germination, and in different tissues of the mature plant, and transcription was also ontogenetically regulated. Heterologous over-expression of Tr-KPI1, Tr-KPI2, Tr-KPI4 and Tr-KPI5 in Nicotiana tabacum retarded larval growth of the herbivore Spodoptera litura, and an increase in the transcription of the pathogenesis-related genes PR1 and PR4 was observed in the Tr-KPI1 and Tr-KPI4 over-expressing lines. RNA interference (RNAi) knock-down lines in white clover displayed significantly altered vegetative growth phenotypes with inhibition of shoot growth and a stimulation of root growth, while knock-down of Tr-KPI1, Tr-KPI2 and Tr-KPI5 transcript abundance also retarded larval growth of S. litura. Examination of these RNAi lines revealed constitutive stress-associated phenotypes as well as altered transcription of cellular signalling genes. These results reveal a functional redundancy across members of the KPI gene family. Further, the regulation of transcription of at least one member of the family, Tr-KPI2, may occupy a central role in the maintenance of a cellular homeostasis. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

  16. Contribution of liver mitochondrial membrane-bound glutathione transferase to mitochondrial permeability transition pores

    International Nuclear Information System (INIS)

    Hossain, Quazi Sohel; Ulziikhishig, Enkhbaatar; Lee, Kang Kwang; Yamamoto, Hideyuki; Aniya, Yoko

    2009-01-01

    We recently reported that the glutathione transferase in rat liver mitochondrial membranes (mtMGST1) is activated by S-glutathionylation and the activated mtMGST1 contributes to the mitochondrial permeability transition (MPT) pore and cytochrome c release from mitochondria [Lee, K.K., Shimoji, M., Quazi, S.H., Sunakawa, H., Aniya, Y., 2008. Novel function of glutathione transferase in rat liver mitochondrial membrane: role for cytochrome c release from mitochondria. Toxcol. Appl. Pharmacol. 232, 109-118]. In the present study we investigated the effect of reactive oxygen species (ROS), generator gallic acid (GA) and GST inhibitors on mtMGST1 and the MPT. When rat liver mitochondria were incubated with GA, mtMGST1 activity was increased to about 3 fold and the increase was inhibited with antioxidant enzymes and singlet oxygen quenchers including 1,4-diazabicyclo [2,2,2] octane (DABCO). GA-mediated mtMGST1 activation was prevented by GST inhibitors such as tannic acid, hematin, and cibacron blue and also by cyclosporin A (CsA). In addition, GA induced the mitochondrial swelling which was also inhibited by GST inhibitors, but not by MPT inhibitors CsA, ADP, and bongkrekic acid. GA also released cytochrome c from the mitochondria which was inhibited completely by DABCO, moderately by GST inhibitors, and somewhat by CsA. Ca 2+ -mediated mitochondrial swelling and cytochrome c release were inhibited by MPT inhibitors but not by GST inhibitors. When the outer mitochondrial membrane was isolated after treatment of mitochondria with GA, mtMGST1 activity was markedly increased and oligomer/aggregate of mtMGST1 was observed. These results indicate that mtMGST1 in the outer mitochondrial membrane is activated by GA through thiol oxidation leading to protein oligomerization/aggregation, which may contribute to the formation of ROS-mediated, CsA-insensitive MPT pore, suggesting a novel mechanism for regulation of the MPT by mtMGST1

  17. Inhibition of the recombinant cattle tick Rhipicephalus (Boophilus) annulatus glutathione S-transferase.

    Science.gov (United States)

    Guneidy, Rasha A; Shahein, Yasser E; Abouelella, Amira M K; Zaki, Eman R; Hamed, Ragaa R

    2014-09-01

    Rhipicephalus (Boophilus) annulatus is a bloodsucking ectoparasite that causes severe production losses in the cattle industry. This study aims to evaluate the in vitro effects of tannic acid, hematin (GST inhibitors) and different plant extracts (rich in tannic acid) on the activity of the recombinant glutathione S-transferase enzyme of the Egyptian cattle tick R. annulatus (rRaGST), in order to confirm their ability to inhibit the parasitic essential detoxification enzyme glutathione S-transferase. Extraction with 70% ethanol of Hibiscus cannabinus (kenaf flowers), Punica granatum (red and white pomegranate peel), Musa acuminata (banana peel) (Musaceae), Medicago sativa (alfalfa seeds), Tamarindus indicus (seed) and Cuminum cyminum (cumin seed) were used to assess: (i) inhibitory capacities of rRaGST and (ii) their phenolic and flavonoid contents. Ethanol extraction of red pomegranate peel contained the highest content of phenolic compounds (29.95mg gallic acid/g dry tissue) compared to the other studied plant extracts. The highest inhibition activities of rRaGST were obtained with kenaf and red pomegranate peel (P. granatum) extracts with IC50 values of 0.123 and 0.136mg dry tissue/ml, respectively. Tannic acid was the more effective inhibitor of rRaGST with an IC50 value equal to 4.57μM compared to delphinidine-HCl (IC50=14.9±3.1μM). Gossypol had a weak inhibitory effect (IC50=43.7μM), and caffeic acid had almost no effect on tick GST activity. The IC50 values qualify ethacrynic acid as a potent inhibitor of rRaGST activity (IC50=0.034μM). Cibacron blue and hematin showed a considerable inhibition effect on rRaGST activity, and their IC50 values were 0.13μM and 7.5μM, respectively. The activity of rRaGST was highest for CDNB (30.2μmol/min/mg protein). The enzyme had also a peroxidatic activity (the specific activity equals 26.5μmol/min/mg protein). Both tannic acid and hematin inhibited rRaGST activity non-competitively with respect to GSH and

  18. Structural snapshots along the reaction pathway of Yersinia pestis RipA, a putative butyryl-CoA transferase

    Energy Technology Data Exchange (ETDEWEB)

    Torres, Rodrigo; Lan, Benson; Latif, Yama; Chim, Nicholas [UC Irvine, 2212 Natural Sciences I, Irvine, CA 92697 (United States); Goulding, Celia W., E-mail: celia.goulding@uci.edu [UC Irvine, 2212 Natural Sciences I, Irvine, CA 92697 (United States); UC Irvine, 2302 Natural Sciences I, Irvine, CA 92697 (United States)

    2014-04-01

    The crystal structures of Y. pestis RipA mutants were determined to provide insights into the CoA transferase reaction pathway. Yersinia pestis, the causative agent of bubonic plague, is able to survive in both extracellular and intracellular environments within the human host, although its intracellular survival within macrophages is poorly understood. A novel Y. pestis three-gene rip (required for intracellular proliferation) operon, and in particular ripA, has been shown to be essential for survival and replication in interferon γ-induced macrophages. RipA was previously characterized as a putative butyryl-CoA transferase proposed to yield butyrate, a known anti-inflammatory shown to lower macrophage-produced NO levels. RipA belongs to the family I CoA transferases, which share structural homology, a conserved catalytic glutamate which forms a covalent CoA-thioester intermediate and a flexible loop adjacent to the active site known as the G(V/I)G loop. Here, functional and structural analyses of several RipA mutants are presented in an effort to dissect the CoA transferase mechanism of RipA. In particular, E61V, M31G and F60M RipA mutants show increased butyryl-CoA transferase activities when compared with wild-type RipA. Furthermore, the X-ray crystal structures of E61V, M31G and F60M RipA mutants, when compared with the wild-type RipA structure, reveal important conformational changes orchestrated by a conserved acyl-group binding-pocket phenylalanine, Phe85, and the G(V/I)G loop. Binary structures of M31G RipA and F60M RipA with two distinct CoA substrate conformations are also presented. Taken together, these data provide CoA transferase reaction snapshots of an open apo RipA, a closed glutamyl-anhydride intermediate and an open CoA-thioester intermediate. Furthermore, biochemical analyses support essential roles for both the catalytic glutamate and the flexible G(V/I)G loop along the reaction pathway, although further research is required to fully

  19. Structural snapshots along the reaction pathway of Yersinia pestis RipA, a putative butyryl-CoA transferase

    International Nuclear Information System (INIS)

    Torres, Rodrigo; Lan, Benson; Latif, Yama; Chim, Nicholas; Goulding, Celia W.

    2014-01-01

    The crystal structures of Y. pestis RipA mutants were determined to provide insights into the CoA transferase reaction pathway. Yersinia pestis, the causative agent of bubonic plague, is able to survive in both extracellular and intracellular environments within the human host, although its intracellular survival within macrophages is poorly understood. A novel Y. pestis three-gene rip (required for intracellular proliferation) operon, and in particular ripA, has been shown to be essential for survival and replication in interferon γ-induced macrophages. RipA was previously characterized as a putative butyryl-CoA transferase proposed to yield butyrate, a known anti-inflammatory shown to lower macrophage-produced NO levels. RipA belongs to the family I CoA transferases, which share structural homology, a conserved catalytic glutamate which forms a covalent CoA-thioester intermediate and a flexible loop adjacent to the active site known as the G(V/I)G loop. Here, functional and structural analyses of several RipA mutants are presented in an effort to dissect the CoA transferase mechanism of RipA. In particular, E61V, M31G and F60M RipA mutants show increased butyryl-CoA transferase activities when compared with wild-type RipA. Furthermore, the X-ray crystal structures of E61V, M31G and F60M RipA mutants, when compared with the wild-type RipA structure, reveal important conformational changes orchestrated by a conserved acyl-group binding-pocket phenylalanine, Phe85, and the G(V/I)G loop. Binary structures of M31G RipA and F60M RipA with two distinct CoA substrate conformations are also presented. Taken together, these data provide CoA transferase reaction snapshots of an open apo RipA, a closed glutamyl-anhydride intermediate and an open CoA-thioester intermediate. Furthermore, biochemical analyses support essential roles for both the catalytic glutamate and the flexible G(V/I)G loop along the reaction pathway, although further research is required to fully

  20. Expression profiling of selected glutathione transferase genes in Zea mays (L.) seedlings infested with cereal aphids.

    Science.gov (United States)

    Sytykiewicz, Hubert; Chrzanowski, Grzegorz; Czerniewicz, Paweł; Sprawka, Iwona; Łukasik, Iwona; Goławska, Sylwia; Sempruch, Cezary

    2014-01-01

    The purpose of this report was to evaluate the expression patterns of selected glutathione transferase genes (gst1, gst18, gst23 and gst24) in the tissues of two maize (Zea mays L.) varieties (relatively resistant Ambrozja and susceptible Tasty Sweet) that were colonized with oligophagous bird cherry-oat aphid (Rhopalosiphum padi L.) or monophagous grain aphid (Sitobion avenae L.). Simultaneously, insect-triggered generation of superoxide anion radicals (O2•-) in infested Z. mays plants was monitored. Quantified parameters were measured at 1, 2, 4, 8, 24, 48 and 72 h post-initial aphid infestation (hpi) in relation to the non-infested control seedlings. Significant increases in gst transcript amounts were recorded in aphid-stressed plants in comparison to the control seedlings. Maximal enhancement in the expression of the gst genes in aphid-attacked maize plants was found at 8 hpi (gst23) or 24 hpi (gst1, gst18 and gst24) compared to the control. Investigated Z. mays cultivars formed excessive superoxide anion radicals in response to insect treatments, and the highest overproduction of O2•- was noted 4 or 8 h after infestation, depending on the aphid treatment and maize genotype. Importantly, the Ambrozja variety could be characterized as having more profound increments in the levels of gst transcript abundance and O2•- generation in comparison with the Tasty Sweet genotype.

  1. In-vitro effect of flavonoids from Solidago canadensis extract on glutathione S-transferase.

    Science.gov (United States)

    Apáti, Pál; Houghton, Peter J; Kite, Geoffrey; Steventon, Glyn B; Kéry, Agnes

    2006-02-01

    Solidago canadensis is typical of a flavonoid-rich herb and the effect of an aqueous ethanol extract on glutathione-S-transferase (GST) activity using HepG2 cells was compared with those of the flavonol quercetin and its glycosides quercitrin and rutin, found as major constituents. The composition of the extract was determined by HPLC and rutin was found to be the major flavonoidal component of the extract. Total GST activity was assessed using 1-chloro-2,4-dinitrobenzene as a substrate. The glycosides rutin and quercitrin gave dose-dependent increases in GST activity, with a 50% and 24.5% increase at 250 mM, respectively, while the aglycone quercetin inhibited the enzyme by 30% at 250 mM. The total extract of the herb gave an overall dose-dependent increase, the fractions corresponding to the flavonoids showed activating effects while those containing caffeic acid derivatives were inhibitory. The activity observed corresponds to that reported for similar compounds in-vivo using rats, thus the HepG2 cell line could serve as a more satisfactory method of assessing the effects of extracts and compounds on GST.

  2. Terminal deoxynucleotidyl transferase in the diagnosis of leukemia and malignant lymphoma.

    Science.gov (United States)

    Kung, P C; Long, J C; McCaffrey, R P; Ratliff, R L; Harrison, T A; Baltimore, D

    1978-05-01

    Neoplastic cells from 253 patients with leukemia and 46 patients with malignant lymphoma were studied for the presence of terminal deoxynucleotidyl transferase (TdT) by biochemical and fluorescent antibody technics. TdT was detected in circulating blast cells from 73 of 77 patients with acute lymphoblastic leukemia, 24 of 72 patients with chronic myelogenous leukemia examined during the blastic phase of the disorder and in cell suspensions of lymph nodes from nine of nine patients with diffuse lymphoblastic lymphoma. Blast cells from six of 10 patients with acute undifferentiated leukemia were TdT positive, but the enzyme was found in only two of 55 patients with acute myeloblastic leukemia. TdT was not detected in other lymphocytic or granulocytic leukemias or in other types of malignant lymphomas. The fluorescent antibody assay for TdT permits rapid and specific identification of the enzyme in single cells. The TdT assay is clinically useful in confirming the diagnosis of acute lymphoblastic leukemia, evaluating patients with blastic chronic myelogenous leukemia, and distinguishing patients with lymphoblastic lymphoma, whose natural history includes rapid extranodal dissemination, from patients with other poorly differentiated malignant lymphomas.

  3. Lack of Ach1 CoA-Transferase Triggers Apoptosis and Decreases Chronological Lifespan in Yeast

    International Nuclear Information System (INIS)

    Orlandi, Ivan; Casatta, Nadia; Vai, Marina

    2012-01-01

    ACH1 encodes a mitochondrial enzyme of Saccharomyces cerevisiae endowed with CoA-transferase activity. It catalyzes the CoASH transfer from succinyl-CoA to acetate generating acetyl-CoA. It is known that ACH1 inactivation results in growth defects on media containing acetate as a sole carbon and energy source which are particularly severe at low pH. Here, we show that chronological aging ach1Δ cells which accumulate a high amount of extracellular acetic acid display a reduced chronological lifespan. The faster drop of cell survival is completely abrogated by alleviating the acid stress either by a calorie restricted regimen that prevents acetic acid production or by transferring chronologically aging mutant cells to water. Moreover, the short-lived phenotype of ach1Δ cells is accompanied by reactive oxygen species accumulation, severe mitochondrial damage, and an early insurgence of apoptosis. A similar pattern of endogenous severe oxidative stress is observed when ach1Δ cells are cultured using acetic acid as a carbon source under acidic conditions. On the whole, our data provide further evidence of the role of acetic acid as cell-extrinsic mediator of cell death during chronological aging and highlight a primary role of Ach1 enzymatic activity in acetic acid detoxification which is important for mitochondrial functionality.

  4. Serum γ-Glutamyl Transferase Is Inversely Associated with Bone Mineral Density Independently of Alcohol Consumption

    Directory of Open Access Journals (Sweden)

    Han Seok Choi

    2016-03-01

    Full Text Available Backgroundγ-Glutamyl transferase (GGT is a well-known marker of chronic alcohol consumption or hepatobiliary diseases. A number of studies have demonstrated that serum levels of GGT are independently associated with cardiovascular and metabolic disorders. The purpose of this study was to test if serum GGT levels are associated with bone mineral density (BMD in Korean adults.MethodsA total of 462 subjects (289 men and 173 women, who visited Severance Hospital for medical checkup, were included in this study. BMD was measured using dual energy X-ray absorptiometry. Cross-sectional association between serum GGT and BMD was evaluated.ResultsAs serum GGT levels increased from the lowest tertile (tertile 1 to the highest tertile (tertile 3, BMD decreased after adjusting for confounders such as age, body mass index, amount of alcohol consumed, smoking, regular exercise, postmenopausal state (in women, hypertension, diabetes mellitus, and hypercholesterolemia. A multiple linear regression analysis showed a negative association between log-transformed serum GGT levels and BMD. In a multiple logistic regression analysis, tertile 3 of serum GGT level was associated with an increased risk for low bone mass compared to tertile 1 (odds ratio, 2.271; 95% confidence interval, 1.340 to 3.850; P=0.002.ConclusionSerum GGT level was inversely associated with BMD in Korean adults. Further study is necessary to fully elucidate the mechanism of the inverse relationship.

  5. The dyad palindromic glutathione transferase P enhancer binds multiple factors including AP1.

    Science.gov (United States)

    Diccianni, M B; Imagawa, M; Muramatsu, M

    1992-10-11

    Glutathione Transferase P (GST-P) gene expression is dominantly regulated by an upstream enhancer (GPEI) consisting of a dyad of palindromically oriented imperfect TPA (12-O-tetradecanoyl-phorbol-13-acetate)-responsive elements (TRE). GPEI is active in AP1-lacking F9 cells as well in AP1-containing HeLa cells. Despite GPEI's similarity to a TRE, c-jun co-transfection has only a minimal effect on transactivation. Antisense c-jun and c-fos co-transfection experiments further demonstrate the lack of a role for AP1 in GPEI mediated trans-activation in F9 cells, although endogenously present AP1 can influence GPEI in HeLa cells. Co-transfection of delta fosB with c-jun, which forms an inactive c-Jun/delta FosB heterodimer that binds TRE sequences, inhibits GPEI-mediated transcription in AP1-lacking F9 cells as well as AP1-containing HeLa cells. These data suggest novel factor(s) other than AP1 are influencing GPEI. Binding studies reveal multiple nucleoproteins bind to GPEI. These factors are likely responsible for the high level of GPEI-mediated transcription observed in the absence of AP1 and during hepatocarcinogenesis.

  6. Expression Profiling of Selected Glutathione Transferase Genes in Zea mays (L.) Seedlings Infested with Cereal Aphids

    Science.gov (United States)

    Sytykiewicz, Hubert; Chrzanowski, Grzegorz; Czerniewicz, Paweł; Sprawka, Iwona; Łukasik, Iwona; Goławska, Sylwia; Sempruch, Cezary

    2014-01-01

    The purpose of this report was to evaluate the expression patterns of selected glutathione transferase genes (gst1, gst18, gst23 and gst24) in the tissues of two maize (Zea mays L.) varieties (relatively resistant Ambrozja and susceptible Tasty Sweet) that were colonized with oligophagous bird cherry-oat aphid (Rhopalosiphum padi L.) or monophagous grain aphid (Sitobion avenae L.). Simultaneously, insect-triggered generation of superoxide anion radicals (O2 •−) in infested Z. mays plants was monitored. Quantified parameters were measured at 1, 2, 4, 8, 24, 48 and 72 h post-initial aphid infestation (hpi) in relation to the non-infested control seedlings. Significant increases in gst transcript amounts were recorded in aphid-stressed plants in comparison to the control seedlings. Maximal enhancement in the expression of the gst genes in aphid-attacked maize plants was found at 8 hpi (gst23) or 24 hpi (gst1, gst18 and gst24) compared to the control. Investigated Z. mays cultivars formed excessive superoxide anion radicals in response to insect treatments, and the highest overproduction of O2 •− was noted 4 or 8 h after infestation, depending on the aphid treatment and maize genotype. Importantly, the Ambrozja variety could be characterized as having more profound increments in the levels of gst transcript abundance and O2 •− generation in comparison with the Tasty Sweet genotype. PMID:25365518

  7. Prevalence of glutathione S-transferase gene deletions and their effect on sickle cell patients

    Directory of Open Access Journals (Sweden)

    Pandey Sanjay

    2012-01-01

    Full Text Available BACKGROUND: Glutathione S-transferase gene deletions are known detoxification agents and cause oxidative damage. Due to the different pathophysiology of anemia in thalassemia and sickle cell disease, there are significant differences in the pathophysiology of iron overload and iron-related complications in these disorders. OBJECTIVE: The aim of this study was to estimate the frequency of the GSTM1 and GSTT1 genotypes in sickle cell disease patients and their effect on iron status. METHODS: Forty sickle cell anemia and sixty sickle ß-thalassemia patients and 100 controls were evaluated to determine the frequency of GST gene deletions. Complete blood counts were performed by an automated cell analyzer. Hemoglobin F, hemoglobin A, hemoglobin A2 and hemoglobin S were measured and diagnosis of patients was achieved by high performance liquid chromatography with DNA extraction by the phenol-chloroform method. The GST null genotype was determined using multiplex polymerase chain reaction and serum ferritin was measured using an ELISA kit. Statistical analysis was by EpiInfo and GraphPad statistics software. RESULTS: An increased frequency of the GSTT1 null genotype (p-value = 0.05 was seen in the patients. The mean serum ferritin level was higher in patients with the GST genotypes than in controls; this was statistically significant for all genotypes except GSTM1, however the higher levels of serum ferritin were due to blood transfusions in patients. CONCLUSION: GST deletions do not play a direct role in iron overload of sickle cell patients.

  8. Characterization and evolutionary implications of the triad Asp-Xxx-Glu in group II phosphopantetheinyl transferases.

    Science.gov (United States)

    Wang, Yue-Yue; Li, Yu-Dong; Liu, Jian-Bo; Ran, Xin-Xin; Guo, Yuan-Yang; Ren, Ni-Ni; Chen, Xin; Jiang, Hui; Li, Yong-Quan

    2014-01-01

    Phosphopantetheinyl transferases (PPTases), which play an essential role in both primary and secondary metabolism, are magnesium binding enzymes. In this study, we characterized the magnesium binding residues of all known group II PPTases by biochemical and evolutionary analysis. Our results suggested that group II PPTases could be classified into two subgroups, two-magnesium-binding-residue-PPTases containing the triad Asp-Xxx-Glu and three-magnesium-binding-residue-PPTases containing the triad Asp-Glu-Glu. Mutations of two three-magnesium-binding-residue-PPTases and one two-magnesium-binding-residue-PPTase indicate that the first and the third residues in the triads are essential to activities; the second residues in the triads are non-essential. Although variations of the second residues in the triad Asp-Xxx-Glu exist throughout the whole phylogenetic tree, the second residues are conserved in animals, plants, algae, and most prokaryotes, respectively. Evolutionary analysis suggests that: the animal group II PPTases may originate from one common ancestor; the plant two-magnesium-binding-residue-PPTases may originate from one common ancestor; the plant three-magnesium-binding-residue-PPTases may derive from horizontal gene transfer from prokaryotes.

  9. Characterization of glutathione S-transferase and its immunodiagnostic potential for detecting Taenia multiceps.

    Science.gov (United States)

    Sun, Ying; Wang, Yu; Huang, Xing; Gu, Xiaobing; Lai, Weimin; Peng, Xuerong; Yang, Guangyou

    2017-08-15

    Taenia multiceps is a widespread zoonotic tapeworm parasite which infects cloven-hoofed animals around the world. Animal infection with Coenurus cerebralis, the coenurus larvae of T. multiceps (Tm), is often fatal, which is a major cause of economic losses in stockbreeding. This study amplified the glutathione S-transferase (GST) gene from the total RNA of C. cerebralis. The resulting protein, Tm-GST, consisted of 201 amino acids, and had a predicted molecular mass of 23.1kDa. Its amino acid sequence shares 77.61% similarity with Echinococcus granulosus GST. Recombinant Tm-GST (rTm-GST) was expressed in Escherichia coli. The protein reacted with serum from goats infected with T. multiceps. Immunofluorescence signals indicated that Tm-GST was largely localized in the parenchymatous area of adult T. multiceps; in addition, it was also apparent in the coenurus. An enzyme-linked immunosorbent assay based on rTm-GST showed specificity of 92.8% (13/14) and sensitivity of 90% (18/20) in detecting anti-GST antibodies in serum from naturally infected animals. This study suggests that Tm-GST has the potential to be used as a diagnostic antigen for Coenurosis. Copyright © 2017. Published by Elsevier B.V.

  10. A glutathione S-transferase gene associated with antioxidant properties isolated from Apis cerana cerana

    Science.gov (United States)

    Liu, Shuchang; Liu, Feng; Jia, Haihong; Yan, Yan; Wang, Hongfang; Guo, Xingqi; Xu, Baohua

    2016-06-01

    Glutathione S-transferases (GSTs) are an important family of multifunctional enzymes in aerobic organisms. They play a crucial role in the detoxification of exogenous compounds, especially insecticides, and protection against oxidative stress. Most previous studies of GSTs in insects have largely focused on their role in insecticide resistance. Here, we isolated a theta class GST gene designated AccGSTT1 from Apis cerana cerana and aimed to explore its antioxidant and antibacterial attributes. Analyses of homology and phylogenetic relationships suggested that the predicted amino acid sequence of AccGSTT1 shares a high level of identity with the other hymenopteran GSTs and that it was conserved during evolution. Quantitative real-time PCR showed that AccGSTT1 is most highly expressed in adult stages and that the expression profile of this gene is significantly altered in response to various abiotic stresses. These results were confirmed using western blot analysis. Additionally, a disc diffusion assay showed that a recombinant AccGSTT1 protein may be roughly capable of inhibiting bacterial growth and that it reduces the resistance of Escherichia coli cells to multiple adverse stresses. Taken together, these data indicate that AccGSTT1 may play an important role in antioxidant processes under adverse stress conditions.

  11. Glutathione S-Transferases: Role in Combating Abiotic Stresses Including Arsenic Detoxification in Plants

    Directory of Open Access Journals (Sweden)

    Smita Kumar

    2018-06-01

    Full Text Available Arsenic (As, naturally occurring metalloid and a potential hazardous material, is found in low concentrations in the environment and emerges from natural sources and anthropogenic activities. The presence of As in ground water, which is used for irrigation, is a matter of great concern since it affects crop productivity and contaminates food chain. In plants, As alters various metabolic pathways in cells including the interaction of substrates/enzymes with the sulfhydryl groups of proteins and the replacement of phosphate in ATP for energy. In addition, As stimulates the generation of free radicals and reactive oxygen species (ROS, resulting in oxidative stress. Glutathione S-transferases (GSTs quench reactive molecules with the addition of glutathione (GSH and protect the cell from oxidative damage. GSTs are a multigene family of isozymes, known to catalyze the conjugation of GSH to miscellany of electrophilic and hydrophobic substrates. GSTs have been reported to be associated with plant developmental processes and are responsive to multitude of stressors. In past, several studies suggested involvement of plant GST gene family in As response due to the requirement of sulfur and GSH in the detoxification of this toxic metalloid. This review provides updated information about the role of GSTs in abiotic and biotic stresses with an emphasis on As uptake, metabolism, and detoxification in plants. Further, the genetic manipulations that helped in enhancing the understanding of the function of GSTs in abiotic stress response and heavy metal detoxification has been reviewed.

  12. The Biochemistry of O-GlcNAc Transferase: Which Functions Make It Essential in Mammalian Cells?

    Science.gov (United States)

    Levine, Zebulon G; Walker, Suzanne

    2016-06-02

    O-linked N-acetylglucosamine transferase (OGT) is found in all metazoans and plays an important role in development but at the single-cell level is only essential in dividing mammalian cells. Postmitotic mammalian cells and cells of invertebrates such as Caenorhabditis elegans and Drosophila can survive without copies of OGT. Why OGT is required in dividing mammalian cells but not in other cells remains unknown. OGT has multiple biochemical activities. Beyond its well-known role in adding β-O-GlcNAc to serine and threonine residues of nuclear and cytoplasmic proteins, OGT also acts as a protease in the maturation of the cell cycle regulator host cell factor 1 (HCF-1) and serves as an integral member of several protein complexes, many of them linked to gene expression. In this review, we summarize current understanding of the mechanisms underlying OGT's biochemical activities and address whether known functions of OGT could be related to its essential role in dividing mammalian cells.

  13. Solution Structural Studies of GTP:Adenosylcobinamide-Phosphateguanylyl Transferase (CobY from Methanocaldococcus jannaschii.

    Directory of Open Access Journals (Sweden)

    Kiran K Singarapu

    Full Text Available GTP:adenosylcobinamide-phosphate (AdoCbi-P guanylyl transferase (CobY is an enzyme that transfers the GMP moiety of GTP to AdoCbi yielding AdoCbi-GDP in the late steps of the assembly of Ado-cobamides in archaea. The failure of repeated attempts to crystallize ligand-free (apo CobY prompted us to explore its 3D structure by solution NMR spectroscopy. As reported here, the solution structure has a mixed α/β fold consisting of seven β-strands and five α-helices, which is very similar to a Rossmann fold. Titration of apo-CobY with GTP resulted in large changes in amide proton chemical shifts that indicated major structural perturbations upon complex formation. However, the CobY:GTP complex as followed by 1H-15N HSQC spectra was found to be unstable over time: GTP hydrolyzed and the protein converted slowly to a species with an NMR spectrum similar to that of apo-CobY. The variant CobYG153D, whose GTP complex was studied by X-ray crystallography, yielded NMR spectra similar to those of wild-type CobY in both its apo- state and in complex with GTP. The CobYG153D:GTP complex was also found to be unstable over time.

  14. [Synthesis of vitamin K2 by isopentenyl transferase NovA in Pichia pastoris Gpn12].

    Science.gov (United States)

    Wu, Xihua; Li, Zhemin; Liu, Hui; Wang, Peng; Wang, Li; Fang, Xue; Sun, Xiaowen; Ni, Wenfeng; Yang, Qiang; Zheng, Zhiming; Zhao, Genhai

    2018-01-25

    The effect of methanol addition on the heterologous expression of isoprenyl transferase NovQ was studied in Pichia pastoris Gpn12, with menadione and isopentenol as precursors to catalyze vitamin K2 (MK-3) synthesis. The expression of NovQ increased by 36% when 2% methanol was added every 24 h. The influence of initial pH, temperature, methanol addition, precursors (menadione, isopentenol) addition, catalytic time and cetyltrimethyl-ammonium bromide (CTAB) addition were explored in the P. pastoris whole-cell catalytic synthesis process of MK-3 in shaking flask. Three significant factors were then studied by response surface method. The optimal catalytic conditions obtained were as follows: catalytic temperature 31.56 ℃, menadione 295.54 mg/L, catalytic time 15.87 h. Consistent with the response surface prediction results, the optimized yield of MK-3 reached 98.47 mg/L in shaking flask, 35% higher than that of the control group. On this basis, the production in a 30-L fermenter reached 189.67 mg/L when the cell catalyst of 220 g/L (dry weight) was used to catalyze the synthesis for 24 h. This method laid the foundation for the large-scale production of MK-3 by P. pastoris Gpn12.

  15. The TCA cycle transferase DLST is important for MYC-mediated leukemogenesis.

    Science.gov (United States)

    Anderson, N M; Li, D; Peng, H L; Laroche, F J F; Mansour, M R; Gjini, E; Aioub, M; Helman, D J; Roderick, J E; Cheng, T; Harrold, I; Samaha, Y; Meng, L; Amsterdam, A; Neuberg, D S; Denton, T T; Sanda, T; Kelliher, M A; Singh, A; Look, A T; Feng, H

    2016-06-01

    Despite the pivotal role of MYC in the pathogenesis of T-cell acute lymphoblastic leukemia (T-ALL) and many other cancers, the mechanisms underlying MYC-mediated tumorigenesis remain inadequately understood. Here we utilized a well-characterized zebrafish model of Myc-induced T-ALL for genetic studies to identify novel genes contributing to disease onset. We found that heterozygous inactivation of a tricarboxylic acid (TCA) cycle enzyme, dihydrolipoamide S-succinyltransferase (Dlst), significantly delayed tumor onset in zebrafish without detectable effects on fish development. DLST is the E2 transferase of the α-ketoglutarate (α-KG) dehydrogenase complex (KGDHC), which converts α-KG to succinyl-CoA in the TCA cycle. RNAi knockdown of DLST led to decreased cell viability and induction of apoptosis in human T-ALL cell lines. Polar metabolomics profiling revealed that the TCA cycle was disrupted by DLST knockdown in human T-ALL cells, as demonstrated by an accumulation of α-KG and a decrease of succinyl-CoA. Addition of succinate, the downstream TCA cycle intermediate, to human T-ALL cells was sufficient to rescue defects in cell viability caused by DLST inactivation. Together, our studies uncovered an important role for DLST in MYC-mediated leukemogenesis and demonstrated the metabolic dependence of T-lymphoblasts on the TCA cycle, thus providing implications for targeted therapy.

  16. Corneal aldehyde dehydrogenase and glutathione S-transferase activity after excimer laser keratectomy in guinea pigs.

    Science.gov (United States)

    Bilgihan, K; Bilgihan, A; Hasanreisoğlu, B; Turkozkan, N

    1998-03-01

    The free radical balance of the eye may be changed by excimer laser keratectomy. Previous studies have demonstrated that excimer laser keratectomy increases the corneal temperature, decreases the superoxide dismutase activity of the aqueous, and induces lipid peroxidation in the superficial corneal stroma. Aldehyde dehydrogenase (ALDH) and glutathione S-transferase (GST) are known to play an important role in corneal metabolism, particularly in detoxification of aldehydes, which are generated from free radical reactions. In three groups of guinea pigs mechanical corneal de-epithelialisation was performed in group I, superficial corneal photoablation in group II, and deep corneal photoablation in group III, and the corneal ALDH and GST activities measured after 48 hours. The mean ALDH and GST activities of group I and II showed no differences compared with the controls (p > 0.05). The corneal ALDH activities were found to be significantly decreased (p < 0.05) and GST activities increased (p < 0.05) in group III. These results suggest that excimer laser treatment of high myopia may change the ALDH and GST activities, metabolism, and free radical balance of the cornea.

  17. Isozyme-specific fluorescent inhibitor of glutathione s-transferase omega 1.

    Science.gov (United States)

    Son, Junghyun; Lee, Jae-Jung; Lee, Jun-Seok; Schüller, Andreas; Chang, Young-Tae

    2010-05-21

    Recently, the glutathione S-transferase omega 1 (GSTO1) is suspected to be involved in certain cancers and neurodegenerative diseases. However, profound investigation on the pathological roles of GSTO1 has been hampered by the lack of specific methods to determine or modulate its activity in biological systems containing other isoforms with similar catalytic function. Here, we report a fluorescent compound that is able to inhibit and monitor the activity of GSTO1. We screened 43 fluorescent chemicals and found a compound (6) that binds specifically to the active site of GSTO1. We observed that compound 6 inhibits GSTO1 by covalent modification but spares other isoforms in HEK293 cells and demonstrated that compound 6 could report the activity of GSTO1 in NIH/3T3 or HEK293 cells by measuring the fluorescence intensity of the labeled amount of GSTO1 in SDS-PAGE. Compound 6 is a useful tool to study GSTO1, applicable as a specific inhibitor and an activity reporter.

  18. Identification and characterisation of multiple glutathione S-transferase genes from the diamondback moth, Plutella xylostella.

    Science.gov (United States)

    Chen, Xi'en; Zhang, Ya-lin

    2015-04-01

    The diamondback moth (DBM), Plutella xylostella, is one of the most harmful insect pests on crucifer crops worldwide. In this study, 19 cDNAs encoding glutathione S-transferases (GSTs) were identified from the genomic and transcriptomic database for DBM (KONAGAbase) and further characterized. Phylogenetic analysis showed that the 19 GSTs were classified into six different cytosolic classes, including four in delta, six in epsilon, three in omega, two in sigma, one in theta and one in zeta. Two GSTs were unclassified. RT-PCR analysis revealed that most GST genes were expressed in all developmental stages, with higher expression in the larval stages. Six DBM GSTs were expressed at the highest levels in the midgut tissue. Twelve purified recombinant GSTs showed varied enzymatic properties towards 1-chloro-2,4-dinitrobenzene and glutathione, whereas rPxGSTo2, rPxGSTz1 and rPxGSTu2 had no activity. Real-time quantitative PCR revealed that expression levels of the 19 DBM GST genes were varied and changed after exposure to acephate, indoxacarb, beta-cypermethrin and spinosad. PxGSTd3 was significantly overexpressed, while PxGSTe3 and PxGSTs2 were significantly downregulated by all four insecticide exposures. The changes in DBM GST gene expression levels exposed to different insecticides indicate that they may play individual roles in tolerance to insecticides and xenobiotics. © 2014 Society of Chemical Industry.

  19. Novel derivatives of aclacinomycin A block cancer cell migration through inhibition of farnesyl transferase.

    Science.gov (United States)

    Magi, Shigeyuki; Shitara, Tetsuo; Takemoto, Yasushi; Sawada, Masato; Kitagawa, Mitsuhiro; Tashiro, Etsu; Takahashi, Yoshikazu; Imoto, Masaya

    2013-03-01

    In the course of screening for an inhibitor of farnesyl transferase (FTase), we identified two compounds, N-benzyl-aclacinomycin A (ACM) and N-allyl-ACM, which are new derivatives of ACM. N-benzyl-ACM and N-allyl-ACM inhibited FTase activity with IC50 values of 0.86 and 2.93 μM, respectively. Not only ACM but also C-10 epimers of each ACM derivative failed to inhibit FTase. The inhibition of FTase by N-benzyl-ACM and N-allyl-ACM seems to be specific, because these two compounds did not inhibit geranylgeranyltransferase or geranylgeranyl pyrophosphate (GGPP) synthase up to 100 μM. In cultured A431 cells, N-benzyl-ACM and N-allyl-ACM also blocked both the membrane localization of H-Ras and activation of the H-Ras-dependent PI3K/Akt pathway. In addition, they inhibited epidermal growth factor (EGF)-induced migration of A431 cells. Thus, N-benzyl-ACM and N-allyl-ACM inhibited EGF-induced migration of A431 cells by inhibiting the farnesylation of H-Ras and subsequent H-Ras-dependent activation of the PI3K/Akt pathway.

  20. Electrochemical DNA biosensor based on grafting-to mode of terminal deoxynucleoside transferase-mediated extension.

    Science.gov (United States)

    Chen, Jinyuan; Liu, Zhoujie; Peng, Huaping; Zheng, Yanjie; Lin, Zhen; Liu, Ailin; Chen, Wei; Lin, Xinhua

    2017-12-15

    Previously reported electrochemical DNA biosensors based on in-situ polymerization approach reveal that terminal deoxynucleoside transferase (TdTase) has good amplifying performance and promising application in the design of electrochemical DNA biosensor. However, this method, in which the background is significantly affected by the amount of TdTase, suffers from being easy to produce false positive result and poor stability. Herein, we firstly present a novel electrochemical DNA biosensor based on grafting-to mode of TdTase-mediated extension, in which DNA targets are polymerized in homogeneous solution and then hybridized with DNA probes on BSA-based DNA carrier platform. It is surprising to find that the background in the grafting-to mode of TdTase-based electrochemical DNA biosensor have little interference from the employed TdTase. Most importantly, the proposed electrochemical DNA biosensor shows greatly improved detection performance over the in-situ polymerization approach-based electrochemical DNA biosensor. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Relation of serum γ-glutamyl transferase activity with copper in an adult population.

    Science.gov (United States)

    Peng, You-Fan; Wang, Chun-Fang; Pan, Guo-Gang

    2017-10-26

    The aim of this study was to evaluate the relationship between serum γ-glutamyl transferase (γ-GGT) activity and serum copper in an adult population. We analyzed 281 adult subjects who regularly attended the physical examination center at the Affiliated Hospital of Youjiang Medical University for Nationalities. The demographic and laboratory data of the participants were divided into two groups according to the median of serum γ-GGT activity. Serum copper concentrations in individuals with higher γ-GGT levels were significantly increased compared with those with lower γ-GGT concentrations (9.9±2.41 vs. 11.2±3.36 μmol/L, pcopper in all eligible subjects (r=0.198, p=0.001). Further, serum γ-GGT maintained a positive correlation with serum copper in both males and females (r=0.322, pcopper after adjusting for multiple potential confounders (b=0.464, p=0.001). This study suggests that serum γ-GGT activity is correlated with copper in the study population, indicating that serum γ-GGT may be a biomarker to evaluate serum copper levels in an adult population.

  2. Legionella shows a diverse secondary metabolism dependent on a broad spectrum Sfp-type phosphopantetheinyl transferase

    Directory of Open Access Journals (Sweden)

    Nicholas J. Tobias

    2016-11-01

    Full Text Available Several members of the genus Legionella cause Legionnaires’ disease, a potentially debilitating form of pneumonia. Studies frequently focus on the abundant number of virulence factors present in this genus. However, what is often overlooked is the role of secondary metabolites from Legionella. Following whole genome sequencing, we assembled and annotated the Legionella parisiensis DSM 19216 genome. Together with 14 other members of the Legionella, we performed comparative genomics and analysed the secondary metabolite potential of each strain. We found that Legionella contains a huge variety of biosynthetic gene clusters (BGCs that are potentially making a significant number of novel natural products with undefined function. Surprisingly, only a single Sfp-like phosphopantetheinyl transferase is found in all Legionella strains analyzed that might be responsible for the activation of all carrier proteins in primary (fatty acid biosynthesis and secondary metabolism (polyketide and non-ribosomal peptide synthesis. Using conserved active site motifs, we predict some novel compounds that are probably involved in cell-cell communication, differing to known communication systems. We identify several gene clusters, which may represent novel signaling mechanisms and demonstrate the natural product potential of Legionella.

  3. Glutathione S-transferase P influences redox and migration pathways in bone marrow.

    Directory of Open Access Journals (Sweden)

    Jie Zhang

    Full Text Available To interrogate why redox homeostasis and glutathione S-transferase P (GSTP are important in regulating bone marrow cell proliferation and migration, we isolated crude bone marrow, lineage negative and bone marrow derived-dendritic cells (BMDDCs from both wild type (WT and knockout (Gstp1/p2(-/- mice. Comparison of the two strains showed distinct thiol expression patterns. WT had higher baseline and reactive oxygen species-induced levels of S-glutathionylated proteins, some of which (sarco-endoplasmic reticulum Ca2(+-ATPase regulate Ca(2+ fluxes and subsequently influence proliferation and migration. Redox status is also a crucial determinant in the regulation of the chemokine system. CXCL12 chemotactic response was stronger in WT cells, with commensurate alterations in plasma membrane polarization/permeability and intracellular calcium fluxes; activities of the downstream kinases, ERK and Akt were also higher in WT. In addition, expression levels of the chemokine receptor CXCR4 and its associated phosphatase, SHP-2, were higher in WT. Inhibition of CXCR4 or SHP2 decreased the extent of CXCL12-induced migration in WT BMDDCs. The differential surface densities of CXCR4, SHP-2 and inositol trisphosphate receptor in WT and Gstp1/p2(-/- cells correlated with the differential CXCR4 functional activities, as measured by the extent of chemokine-induced directional migration and differences in intracellular signaling. These observed differences contribute to our understanding of how genetic ablation of GSTP causes different levels of myeloproliferation and migration [corrected

  4. Recognition and Detoxification of the Insecticide DDT by Drosophila melanogaster Glutathione S-Transferase D1

    Energy Technology Data Exchange (ETDEWEB)

    Low, Wai Yee; Feil, Susanne C.; Ng, Hooi Ling; Gorman, Michael A.; Morton, Craig J.; Pyke, James; McConville, Malcolm J.; Bieri, Michael; Mok, Yee-Foong; Robin, Charles; Gooley, Paul R.; Parker, Michael W.; Batterham, Philip (SVIMR-A); (Melbourne)

    2010-06-14

    GSTD1 is one of several insect glutathione S-transferases capable of metabolizing the insecticide DDT. Here we use crystallography and NMR to elucidate the binding of DDT and glutathione to GSTD1. The crystal structure of Drosophila melanogaster GSTD1 has been determined to 1.1 {angstrom} resolution, which reveals that the enzyme adopts the canonical GST fold but with a partially occluded active site caused by the packing of a C-terminal helix against one wall of the binding site for substrates. This helix would need to unwind or be displaced to enable catalysis. When the C-terminal helix is removed from the model of the crystal structure, DDT can be computationally docked into the active site in an orientation favoring catalysis. Two-dimensional {sup 1}H,{sup 15}N heteronuclear single-quantum coherence NMR experiments of GSTD1 indicate that conformational changes occur upon glutathione and DDT binding and the residues that broaden upon DDT binding support the predicted binding site. We also show that the ancestral GSTD1 is likely to have possessed DDT dehydrochlorinase activity because both GSTD1 from D. melanogaster and its sibling species, Drosophila simulans, have this activity.

  5. Targeting Glutathione-S Transferase Enzymes in Musculoskeletal Sarcomas: A Promising Therapeutic Strategy

    Directory of Open Access Journals (Sweden)

    Michela Pasello

    2011-01-01

    Full Text Available Recent studies have indicated that targeting glutathione-S-transferase (GST isoenzymes may be a promising novel strategy to improve the efficacy of conventional chemotherapy in the three most common musculoskeletal tumours: osteosarcoma, Ewing's sarcoma, and rhabdomyosarcoma. By using a panel of 15 drug-sensitive and drug-resistant human osteosarcoma, Ewing's sarcoma, and rhabdomyosarcoma cell lines, the efficay of the GST-targeting agent 6-(7-nitro-2,1,3-benzoxadiazol-4-ylthiohexanol (NBDHEX has been assessed and related to GST isoenzymes expression (namely GSTP1, GSTA1, GSTM1, and MGST. NBDHEX showed a relevant in vitro activity on all cell lines, including the drug-resistant ones and those with higher GSTs levels. The in vitro activity of NBDHEX was mostly related to cytostatic effects, with a less evident apoptotic induction. NBDHEX positively interacted with doxorubicin, vincristine, cisplatin but showed antagonistic effects with methotrexate. In vivo studies confirmed the cytostatic efficay of NBDHEX and its positive interaction with vincristine in Ewing's sarcoma cells, and also indicated a positive effect against the metastatisation of osteosarcoma cells. The whole body of evidence found in this study indicated that targeting GSTs in osteosarcoma, Ewing's sarcoma and rhabdomyosarcoma may be an interesting new therapeutic option, which can be considered for patients who are scarcely responsive to conventional regimens.

  6. Terminal deoxynucleotidyl transferase activity in the regenerating thymus of X-irradiated mice

    International Nuclear Information System (INIS)

    Daculsi, R.; Astier, T.; Legrand, E.; Duplan, J.F.

    1982-01-01

    The distribution of terminal deoyxnucleotidyl transferase (TdT) enzyme activity (EU per 10 8 cells) between peaks I and II was followed for a period of 42 days in regenerating thymus of lethally irradiated (9 Gy) C3H mice restored with 10 6 (C3H x AKR) F1 bone marrow cells. The detection of Thy-1.1 and Thy-1.2 surface antigens allowed for the discrimination between host and donor cells, and the main subpopulations of thymic cells were characterized by their sensitivity to H-2sup(k) antiserum and to dexamethazone. Two peaks of TdT activity could be detected on phosphocellulose chromatographic separation. The distribution of TdT activity between these two peaks was followed during the two periods of thymic endo- and exoregeneration. Peak I TdT activity was closely correlated with the variation in the percentage of the high H-2 population. Peak II activity was mostly related to low H-2 cells. The per cell content of both peak I and peak II activities exceeded the norm in rapidly expanding populations. Finally between days 10 and 14 the TdT activity of the endoregenerating population was apparently not different from that of the exoregenerating population between days 14 and 22. (Auth.)

  7. The pleuromutilin drugs tiamulin and valnemulin bind to the RNA at the peptidyl transferase centre on the ribosome

    DEFF Research Database (Denmark)

    Poulsen, S M; Karlsson, M; Johansson, L B

    2001-01-01

    The pleuromutilin antibiotic derivatives, tiamulin and valnemulin, inhibit protein synthesis by binding to the 50S ribosomal subunit of bacteria. The action and binding site of tiamulin and valnemulin was further characterized on Escherichia coli ribosomes. It was revealed that these drugs...... centre and have been associated with binding of several antibiotics. Competitive footprinting shows that tiamulin and valnemulin can bind concurrently with the macrolide erythromycin but compete with the macrolide carbomycin, which is a peptidyl transferase inhibitor. We infer from these and previous...... results that tiamulin and valnemulin interact with the rRNA in the peptidyl transferase slot on the ribosomes in which they prevent the correct positioning of the CCA-ends of tRNAs for peptide transfer....

  8. A model to environmental monitoring based on glutathione-S-transferase activity and branchial lesions in catfish

    Science.gov (United States)

    Neta, Raimunda Nonata Fortes Carvalho; Torres, Audalio Rebelo

    2017-11-01

    In this work, we validate the glutathione-S-transferase and branchial lesions as biomarkers in catfish Sciades herzbergii to obtain a predictive model of the environmental impact effects in a harbor of Brazil. The catfish were sampled from a port known to be contaminated with heavy metals and organic compounds and from a natural reserve in São Marcos Bay, Maranhão. Two biomarkers, hepatic glutathione S-transferase (GST) activity and branchial lesions were analyzed. The values for GST activity were modeled with the occurrence of branchial lesions by fitting a third order polynomial. Results from the mathematical model indicate that GST activity has a strong polynomial relationship with the occurrence of branchial lesions in both the wet and the dry seasons, but only at the polluted port site. Our mathematic model indicates that when the GST ceases to act, serious branchial lesions are observed in the catfish of the contaminated port area.

  9. Glutathione S-Transferase of Brown Planthoppers (Nilaparvata lugens) Is Essential for Their Adaptation to Gramine-Containing Host Plants

    OpenAIRE

    Sun, Xiao-Qin; Zhang, Mao-Xin; Yu, Jing-Ya; Jin, Yu; Ling, Bing; Du, Jin-Ping; Li, Gui-Hua; Qin, Qing-Ming; Cai, Qing-Nian

    2013-01-01

    Plants have evolved complex processes to ward off attacks by insects. In parallel, insects have evolved mechanisms to thwart these plant defenses. To gain insight into mechanisms that mediate this arms race between plants and herbivorous insects, we investigated the interactions between gramine, a toxin synthesized by plants of the family Gramineae, and glutathione S transferase (GST), an enzyme found in insects that is known to detoxify xenobiotics. Here, we demonstrate that rice (Oryza sati...

  10. The activity of glutathione S-transferase in hepatopancreas of Procambarus clarkii: seasonal variations and the influence of environmental pollutants.

    Science.gov (United States)

    Nies, E; Almar, M M; Hermenegildo, C; Monsalve, E; Romero, F J

    1991-01-01

    1. The glutathione S-transferase activity in hepatopancreas of the American red crayfish Procambarus clarkii after 15 days' acclimatization in tap water aquaria was measured in specimens collected monthly for a whole year, and shows seasonal variation. 2. Previous data on the environmental pollution of Lake Albufera suggest a possible correlation with the activity tested in the different seasons of the year considering the results of non-acclimatized animals.

  11. β(1,3-glucanosyl-transferase activity is essential for cell wall integrity and viability of Schizosaccharomyces pombe.

    Directory of Open Access Journals (Sweden)

    María de Medina-Redondo

    Full Text Available BACKGROUND: The formation of the cell wall in Schizosaccharomyces pombe requires the coordinated activity of enzymes involved in the biosynthesis and modification of β-glucans. The β(1,3-glucan synthase complex synthesizes linear β(1,3-glucans, which remain unorganized until they are cross-linked to other β(1,3-glucans and other cell wall components. Transferases of the GH72 family play important roles in cell wall assembly and its rearrangement in Saccharomyces cerevisiae and Aspergillus fumigatus. Four genes encoding β(1,3-glucanosyl-transferases -gas1(+, gas2(+, gas4(+ and gas5(+- are present in S. pombe, although their function has not been analyzed. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report the characterization of the catalytic activity of gas1p, gas2p and gas5p together with studies directed to understand their function during vegetative growth. From the functional point of view, gas1p is essential for cell integrity and viability during vegetative growth, since gas1Δ mutants can only grow in osmotically supported media, while gas2p and gas5p play a minor role in cell wall construction. From the biochemical point of view, all of them display β(1,3-glucanosyl-transferase activity, although they differ in their specificity for substrate length, cleavage point and product size. In light of all the above, together with the differences in expression profiles during the life cycle, the S. pombe GH72 proteins may accomplish complementary, non-overlapping functions in fission yeast. CONCLUSIONS/SIGNIFICANCE: We conclude that β(1,3-glucanosyl-transferase activity is essential for viability in fission yeast, being required to maintain cell integrity during vegetative growth.

  12. Forage Quantity and Quality of Berseem Clover (Trifolium ‎alexandrinum L. as Affected by Uses of Pseudomonas putida ‎Strains and Phophorus Fertilizer in the Second Crop

    Directory of Open Access Journals (Sweden)

    Mohammad Hossein Ansari

    2017-05-01

    Full Text Available Effects of phosphate fertilizer and pseudomonas putida strains on the quantity and quality of forage of berseem clover as a second crop was studied in a factorial field experiment using randomized complete block design with three replications at Fooman, Guilan province, Iran. Treatments consisted of phosphate fertilizer with three levels (0, 75 and 150 kg/ha as triple super phosphate and Pseudomonas putida strains with four levels (M21, M5, M168 and control. The results showed that use of phosphate fertilizers increased the soil pH during growing season while bacterial inoculation adjusted soil pH. The bacterial inoculation increased amount of crude protein, digestible protein, acidic and alkaline phosphatase activity compared to non-inoculated treatment, but it decreased crude fiber of the forage. Clover forage yield, protein yield and phosphorus content of foliage also were influenced by the interaction of bacterial strains and phosphate fertilizer. The highest forage and protein yield were obtained by using strain M5+150 kg P ha-1. Significant increases in forage and protein yield were found to be 16.49% and 8.01%, respectively, as compared with non-inoculated treatment. Based on the result of this experiment, application of 150 kg P ha-1 and Pseudomonas putida strain M5 inoculation can be used to obtain highest forage yield and quality of berseem clover as second crop in the experimental site.

  13. Meat consumption, N-acetyl transferase 1 and 2 polymorphism and risk of breast cancer, in Danish postmenopausal women

    DEFF Research Database (Denmark)

    Egeberg, Rikke; Olsen, Anja; Autrup, Herman

    2008-01-01

    The aim of this study was to investigate whether polymorphisms in N-acetyl transferase 1 and 2 modify the association between meat consumption and risk of breast cancer. A nested case-control study was conducted among 24697 postmenopausal women included in the 'Diet, Cancer and Health' cohort study...... (1993-2000). Three hundred and seventy-eight breast cancer cases were identified and matched to 378 controls. The incidence rate ratio (95% confidence interval) for breast cancer was 1.09 (1.02-1.17) for total meat, 1.15 (1.01-1.31) for red meat and 1.23 (1.04-1.45) for processed meat per 25 g daily...... total meat intake and red meat intake and breast cancer risk were confined to intermediate/fast N-acetyl transferase 2 acetylators (P-interaction=0.03 and 0.04). Our findings support an association between meat consumption and breast cancer risk and that N-acetyl transferase 2 polymorphism has...

  14. Structures of a putative ζ-class glutathione S-transferase from the pathogenic fungus Coccidioides immitis

    International Nuclear Information System (INIS)

    Edwards, Thomas E.; Bryan, Cassie M.; Leibly, David J.; Dieterich, Shellie H.; Abendroth, Jan; Sankaran, Banumathi; Sivam, Dhileep; Staker, Bart L.; Van Voorhis, Wesley C.; Myler, Peter J.; Stewart, Lance J.

    2011-01-01

    The pathogenic fungus C. immitis causes coccidioidomycosis, a potentially fatal disease. Here, apo and glutathione-bound crystal structures of a previously uncharacterized protein from C. immitis that appears to be a ζ-class glutathione S-transferase are presented. Coccidioides immitis is a pathogenic fungus populating the southwestern United States and is a causative agent of coccidioidomycosis, sometimes referred to as Valley Fever. Although the genome of this fungus has been sequenced, many operons are not properly annotated. Crystal structures are presented for a putative uncharacterized protein that shares sequence similarity with ζ-class glutathione S-transferases (GSTs) in both apo and glutathione-bound forms. The apo structure reveals a nonsymmetric homodimer with each protomer comprising two subdomains: a C-terminal helical domain and an N-terminal thioredoxin-like domain that is common to all GSTs. Half-site binding is observed in the glutathione-bound form. Considerable movement of some components of the active site relative to the glutathione-free form was observed, indicating an induced-fit mechanism for cofactor binding. The sequence homology, structure and half-site occupancy imply that the protein is a ζ-class glutathione S-transferase, a maleylacetoacetate isomerase (MAAI)

  15. Neuroantibodies (NAB) in African-American Children: Associations with Gender, Glutathione-S-Transferase (GST)Pi Polymorphisms (SNP) and Heavy Metals

    Science.gov (United States)

    CONTACT (NAME ONLY): Hassan El-Fawal Abstract Details PRESENTATION TYPE: Platform or Poster CURRENT CATEGORY: Neurodegenerative Disease | Biomarkers | Neurotoxicity, Metals KEYWORDS: Autoantibodies, Glutathione-S-Transferase, DATE/TIME LAST MODIFIED: DATE/TIME SUBMITTED: Abs...

  16. Evaluation of antioxidant and antimicrobial activity of seaweed ( Sargassum sp.) extract: A study on inhibition of glutathione-S-transferase Activity

    Digital Repository Service at National Institute of Oceanography (India)

    Patra, J.K.; Rath, S.K.; Jena, K.B.; Rathod, V.K.; Thatoi, H.

    In the present study, the free radical scavenging potentials (DPPH radical and hydroxyl radical), inhibition of lipid peroxidation, and glutathione-S-transferase and antimicrobial properties of Sargassum sp. extract were investigated. The tested...

  17. Dairy cow responses to graded levels of rapeseed and soya bean expeller supplementation on a red clover/grass silage-based diet.

    Science.gov (United States)

    Rinne, M; Kuoppala, K; Ahvenjärvi, S; Vanhatalo, A

    2015-12-01

    The effects of rapeseed and soya bean expeller (SBE) supplementation on digestion and milk production responses in dairy cows were investigated in an incomplete Latin square design using five cows and four 3-week periods. The experimental diets consisted of five concentrate treatments fed at a rate of 9 kg/day: a mixture of barley and oats, which was replaced with rapeseed or SBE at two levels (CP concentration (g/kg dry matter (DM)) of 130 for the control concentrate and 180 and 230 for the two protein supplemented levels). A mixture of grass and red clover silage (1:1) was fed ad libitum and it had a CP concentration of 157 g/kg DM. Supply of nutrients to the lower tract was measured using the omasal canal sampling technique, and total digestion from total faecal collection. Protein supplementation increased omasal canal amino acid (AA) flows and plasma concentrations of AA, and was also reflected as increased milk production. However, N use efficiency (NUE) decreased with increased protein supplementation. Rapeseed expeller (RSE) tended to increase silage DM intake and elicited higher milk production responses compared with SBE and also resulted in a higher NUE. The differences between the protein supplements in nitrogen metabolism were relatively small, for example, there were no differences in the efficiency of microbial protein synthesis or omasal canal flows of nitrogenous components between them, but plasma methionine concentration was lower for soya bean-fed cows at the high CP level in particular. The lower milk protein production responses to SBE than to RSE supplementation were at least partly caused by increased silage DM and by the lower methionine supply, which may further have been amplified by the use of red clover in the basal diet. Although feed intake, diet digestion, AA supply and milk production were all consistently improved by protein supplementation, there was a simultaneous decrease in NUE. In the current study, the milk protein production

  18. Quantitative and Qualitative Involvement of P3N-PIPO in Overcoming Recessive Resistance against Clover Yellow Vein Virus in Pea Carrying the cyv1 Gene

    Science.gov (United States)

    Choi, Sun Hee; Hagiwara-Komoda, Yuka; Atsumi, Go; Shimada, Ryoko; Hisa, Yusuke; Naito, Satoshi

    2013-01-01

    In pea carrying cyv1, a recessive gene for resistance to Clover yellow vein virus (ClYVV), ClYVV isolate Cl-no30 was restricted to the initially infected cells, whereas isolate 90-1 Br2 overcame this resistance. We mapped the region responsible for breaking of cyv1-mediated resistance by examining infection of cyv1 pea with chimeric viruses constructed from parts of Cl-no30 and 90-1 Br2. The breaking of resistance was attributed to the P3 cistron, which is known to produce two proteins: P3, from the main open reading frame (ORF), and P3N-PIPO, which has the N-terminal part of P3 fused to amino acids encoded by a small open reading frame (ORF) called PIPO in the +2 reading frame. We introduced point mutations that were synonymous with respect to the P3 protein but nonsynonymous with respect to the P3N-PIPO protein, and vice versa, into the chimeric viruses. Infection of plants with these mutant viruses revealed that both P3 and P3N-PIPO were involved in overcoming cyv1-mediated resistance. Moreover, P3N-PIPO quantitatively affected the virulence of Cl-no30 in cyv1 pea. Additional expression in trans of the P3N-PIPO derived from Cl-no30, using White clover mosaic virus as a vector, enabled Cl-no30 to move to systemic leaves in cyv1 pea. Susceptible pea plants infected with chimeric ClYVV possessing the P3 cistron of 90-1 Br2, and which were therefore virulent toward cyv1 pea, accumulated more P3N-PIPO than did those infected with Cl-no30, suggesting that the higher level of P3N-PIPO in infected cells contributed to the breaking of resistance by 90-1 Br2. This is the first report showing that P3N-PIPO is a virulence determinant in plants resistant to a potyvirus. PMID:23616656

  19. Preferential flow, nitrogen transformations and 15N balance under urine-affected areas of irrigated and non-irrigated clover-based pastures

    Science.gov (United States)

    Pakro, Naser; Dillon, Peter

    1995-12-01

    Urine-affected areas can lead to considerable losses of N by leaching, ammonia volatilisation and denitrification from dairy pastures in the southeast of South Australia. Potable groundwater supplies are considered to have become contaminated by nitrate as a result of leaching from these leguminous pastures. Dairy cow urine, labelled with 15N urea, was applied to micro-plots and mini-lysimeters installed in two adjacent irrigated (white clover-rye grass) and non-irrigated (subterranean clover-annual grasses) paddocks of a dairy farm on four occasions representing different seasonal conditions. These experiments allowed measurement of nitrogen transformations, recovery of 15N in the pasture and soil, and leaching below various depths. Gaseous losses were calculated from the nitrogen balance. The results of the four experiments showed that within a day of urine application up to 40% of the applied urinary-N was leached below a depth of 150 mm as a result of macropore flow in the irrigated paddock, and up to 24% in the non-irrigated one. After application to the irrigated paddock 17% of the urinary-N moved immediately below 300 mm but only 2% below the 450-mm depth. The urinary-N remaining in the soil was converted from urea to ammonium within a day regardless of season. Within the first 7 days of application six times more nitrate was produced in summer than in winter. This has obvious implications for leaching potential. Leaching of 15N from the top 150 mm of soil, following urine applications in all seasons, was between 41% and 62% of the applied 15N in the irrigated paddock and 25-51% in the non-irrigated paddock. However, leaching losses measured at depths of 300 or 450 mm were smaller by a factor of 2-4. The leaching loss of 15N applied in spring in both paddocks was 41% below 150 mm and 12% below 450 mm. Recovery of 15N from the soil-plant system in the 450-nm deep lysimeters was ˜60% of that applied. Estimated ammonia was ˜9% of applied 15N with no paddock

  20. Effects of maturity and harvest season of grass-clover silage and of forage-to-concentrate ratio on milk production of dairy cows.

    Science.gov (United States)

    Alstrup, L; Søegaard, K; Weisbjerg, M R

    2016-01-01

    This study examined the effects of maturity and season of harvest of grass-clover silages and forage:concentrate ratio (FCR) on feed intake, milk production, chewing activity, digestibility, and fecal consistency of Holstein dairy cows. Comparison included 2 cuts in spring season (early and late) and 2 cuts in summer season (early and late) combined with high FCR (80:20; HFCR) and low FCR (50:50; LFCR). The experiment included 24 lactating Holstein cows arranged as 2 repeated 4 × 4 Latin squares with four 21-d periods and included measurements of feed composition, feed intake, milk production and composition, chewing activities, digestibilities, and fecal dry matter (DM) concentration and scoring. Forages were fed as two-thirds grass-clover and one-third corn silage supplemented with either 20 or 50% concentrate. Rations were fed ad libitum as total mixed rations. Early maturity cuts were more digestible than late maturity cuts, which was also reflected in a lower concentration of neutral detergent fiber (NDF) in early maturity cuts, whereas summer cuts had a higher crude protein concentration than spring cuts. Increased maturity decreased the intake of DM and energy, increased NDF intake, and decreased the yield of energy-corrected milk (ECM). Summer cuts increased the ECM yield compared with spring cuts. Milk yield (kg and kilogram of ECM) was numerically higher for cows fed early summer cut, independent of FCR in the ration. Milk protein concentration decreased, or tended to decrease, with maturity. For LFCR, the milk fat concentration increased with maturity resulting in a decreased protein:fat ratio. At HFCR, increased maturity increased the time spent chewing per kilogram of DM. Digestibility of silages was positively correlated with the fecal DM concentration. The DM intake and ECM yield showed no significant response to FCR in the ration, but the milk composition was affected. The LFCR decreased the milk fat percentage and increased the milk protein

  1. Glutathione S-transferase P protects against cyclophosphamide-induced cardiotoxicity in mice

    Energy Technology Data Exchange (ETDEWEB)

    Conklin, Daniel J., E-mail: dj.conklin@louisville.edu [Diabetes and Obesity Center, University of Louisville, Louisville, KY 40292 (United States); Institute of Molecular Cardiology, University of Louisville, Louisville, KY 40292 (United States); Haberzettl, Petra; Jagatheesan, Ganapathy; Baba, Shahid [Diabetes and Obesity Center, University of Louisville, Louisville, KY 40292 (United States); Institute of Molecular Cardiology, University of Louisville, Louisville, KY 40292 (United States); Merchant, Michael L. [Diabetes and Obesity Center, University of Louisville, Louisville, KY 40292 (United States); Division of Nephrology, Department of Medicine, University of Louisville, Louisville, KY 40292 (United States); Prough, Russell A. [Diabetes and Obesity Center, University of Louisville, Louisville, KY 40292 (United States); Department of Biochemistry and Molecular Biology, University of Louisville, Louisville, KY 40292 (United States); Williams, Jessica D. [University of Cincinnati College of Medicine, Internal Medicine, Cincinnati, OH 45267 (United States); Prabhu, Sumanth D. [Division of Cardiovascular Disease, University of Alabama-Birmingham, Birmingham, AL 35294 (United States); Bhatnagar, Aruni [Diabetes and Obesity Center, University of Louisville, Louisville, KY 40292 (United States); Institute of Molecular Cardiology, University of Louisville, Louisville, KY 40292 (United States); Department of Biochemistry and Molecular Biology, University of Louisville, Louisville, KY 40292 (United States)

    2015-06-01

    High-dose chemotherapy regimens using cyclophosphamide (CY) are frequently associated with cardiotoxicity that could lead to myocyte damage and congestive heart failure. However, the mechanisms regulating the cardiotoxic effects of CY remain unclear. Because CY is converted to an unsaturated aldehyde acrolein, a toxic, reactive CY metabolite that induces extensive protein modification and myocardial injury, we examined the role of glutathione S-transferase P (GSTP), an acrolein-metabolizing enzyme, in CY cardiotoxicity in wild-type (WT) and GSTP-null mice. Treatment with CY (100–300 mg/kg) increased plasma levels of creatine kinase-MB isoform (CK·MB) and heart-to-body weight ratio to a significantly greater extent in GSTP-null than WT mice. In addition to modest yet significant echocardiographic changes following acute CY-treatment, GSTP insufficiency was associated with greater phosphorylation of c-Jun and p38 as well as greater accumulation of albumin and protein–acrolein adducts in the heart. Mass spectrometric analysis revealed likely prominent modification of albumin, kallikrein-1-related peptidase, myoglobin and transgelin-2 by acrolein in the hearts of CY-treated mice. Treatment with acrolein (low dose, 1–5 mg/kg) also led to increased heart-to-body weight ratio and myocardial contractility changes. Acrolein induced similar hypotension in GSTP-null and WT mice. GSTP-null mice also were more susceptible than WT mice to mortality associated with high-dose acrolein (10–20 mg/kg). Collectively, these results suggest that CY cardiotoxicity is regulated, in part, by GSTP, which prevents CY toxicity by detoxifying acrolein. Thus, humans with low cardiac GSTP levels or polymorphic forms of GSTP with low acrolein-metabolizing capacity may be more sensitive to CY toxicity. - Graphical abstract: Cyclophosphamide (CY) treatment results in P450-mediated metabolic formation of phosphoramide mustard and acrolein (3-propenal). Acrolein is either metabolized and

  2. Glutathione S-transferase P protects against cyclophosphamide-induced cardiotoxicity in mice

    International Nuclear Information System (INIS)

    Conklin, Daniel J.; Haberzettl, Petra; Jagatheesan, Ganapathy; Baba, Shahid; Merchant, Michael L.; Prough, Russell A.; Williams, Jessica D.; Prabhu, Sumanth D.; Bhatnagar, Aruni

    2015-01-01

    High-dose chemotherapy regimens using cyclophosphamide (CY) are frequently associated with cardiotoxicity that could lead to myocyte damage and congestive heart failure. However, the mechanisms regulating the cardiotoxic effects of CY remain unclear. Because CY is converted to an unsaturated aldehyde acrolein, a toxic, reactive CY metabolite that induces extensive protein modification and myocardial injury, we examined the role of glutathione S-transferase P (GSTP), an acrolein-metabolizing enzyme, in CY cardiotoxicity in wild-type (WT) and GSTP-null mice. Treatment with CY (100–300 mg/kg) increased plasma levels of creatine kinase-MB isoform (CK·MB) and heart-to-body weight ratio to a significantly greater extent in GSTP-null than WT mice. In addition to modest yet significant echocardiographic changes following acute CY-treatment, GSTP insufficiency was associated with greater phosphorylation of c-Jun and p38 as well as greater accumulation of albumin and protein–acrolein adducts in the heart. Mass spectrometric analysis revealed likely prominent modification of albumin, kallikrein-1-related peptidase, myoglobin and transgelin-2 by acrolein in the hearts of CY-treated mice. Treatment with acrolein (low dose, 1–5 mg/kg) also led to increased heart-to-body weight ratio and myocardial contractility changes. Acrolein induced similar hypotension in GSTP-null and WT mice. GSTP-null mice also were more susceptible than WT mice to mortality associated with high-dose acrolein (10–20 mg/kg). Collectively, these results suggest that CY cardiotoxicity is regulated, in part, by GSTP, which prevents CY toxicity by detoxifying acrolein. Thus, humans with low cardiac GSTP levels or polymorphic forms of GSTP with low acrolein-metabolizing capacity may be more sensitive to CY toxicity. - Graphical abstract: Cyclophosphamide (CY) treatment results in P450-mediated metabolic formation of phosphoramide mustard and acrolein (3-propenal). Acrolein is either metabolized and

  3. The poplar phi class glutathione transferase: expression, activity and structure of GSTF1

    Directory of Open Access Journals (Sweden)

    Henri ePégeot

    2014-12-01

    Full Text Available Glutathione transferases (GSTs constitute a superfamily of enzymes with essential roles in cellular detoxification and secondary metabolism in plants as in other organisms. Several plant GSTs, including those of the Phi class (GSTFs, require a conserved catalytic serine residue to perform glutathione (GSH-conjugation reactions. Genomic analyses revealed that terrestrial plants have around 10 GSTFs, 8 in the Populus trichocarpa genome, but their physiological functions and substrates are mostly unknown. Transcript expression analyses showed a predominant expression of all genes both in reproductive (female flowers, fruits, floral buds and vegetative organs (leaves, petioles. Here, we show that the recombinant poplar GSTF1 (PttGSTF1 possesses peroxidase activity towards cumene hydroperoxide and GSH-conjugation activity towards model substrates such as 2,4-dinitrochlorobenzene, benzyl and phenetyl isothiocyanate, 4-nitrophenyl butyrate and 4-hydroxy-2-nonenal but interestingly not on previously identified GSTF-class substrates. In accordance to analytical gel filtration data, crystal structure of PttGSTF1 showed a canonical dimeric organization with bound GSH or MES molecules. The structure of these protein-substrate complexes allowed delineating the residues contributing to both the G and H sites that form the active site cavity. In sum, the presence of GSTF1 transcripts and proteins in most poplar organs especially those rich in secondary metabolites such as flowers and fruits, together with its GSH-conjugation activity and its documented stress-responsive expression suggest that its function is associated with the catalytic transformation of metabolites and/or peroxide removal rather than with ligandin properties as previously reported for other GSTFs.

  4. Glutathione S Transferases Polymorphisms Are Independent Prognostic Factors in Lupus Nephritis Treated with Cyclophosphamide.

    Directory of Open Access Journals (Sweden)

    Alexandra Audemard-Verger

    Full Text Available To investigate association between genetic polymorphisms of GST, CYP and renal outcome or occurrence of adverse drug reactions (ADRs in lupus nephritis (LN treated with cyclophosphamide (CYC. CYC, as a pro-drug, requires bioactivation through multiple hepatic cytochrome P450s and glutathione S transferases (GST.We carried out a multicentric retrospective study including 70 patients with proliferative LN treated with CYC. Patients were genotyped for polymorphisms of the CYP2B6, CYP2C19, GSTP1, GSTM1 and GSTT1 genes. Complete remission (CR was defined as proteinuria ≤0.33g/day and serum creatinine ≤124 µmol/l. Partial remission (PR was defined as proteinuria ≤1.5g/day with a 50% decrease of the baseline proteinuria value and serum creatinine no greater than 25% above baseline.Most patients were women (84% and 77% were Caucasian. The mean age at LN diagnosis was 41 ± 10 years. The frequency of patients carrying the GST null genotype GSTT1-, GSTM1-, and the Ile→105Val GSTP1 genotype were respectively 38%, 60% and 44%. In multivariate analysis, the Ile→105Val GSTP1 genotype was an independent factor of poor renal outcome (achievement of CR or PR (OR = 5.01 95% CI [1.02-24.51] and the sole factor that influenced occurrence of ADRs was the GSTM1 null genotype (OR = 3.34 95% CI [1.064-10.58]. No association between polymorphisms of cytochrome P450s gene and efficacy or ADRs was observed.This study suggests that GST polymorphisms highly impact renal outcome and occurrence of ADRs related to CYC in LN patients.

  5. Glutathione-supported arsenate reduction coupled to arsenolysis catalyzed by ornithine carbamoyl transferase

    International Nuclear Information System (INIS)

    Nemeti, Balazs; Gregus, Zoltan

    2009-01-01

    Three cytosolic phosphorolytic/arsenolytic enzymes, (purine nucleoside phosphorylase [PNP], glycogen phosphorylase, glyceraldehyde-3-phosphate dehydrogenase) have been shown to mediate reduction of arsenate (AsV) to the more toxic arsenite (AsIII) in a thiol-dependent manner. With unknown mechanism, hepatic mitochondria also reduce AsV. Mitochondria possess ornithine carbamoyl transferase (OCT), which catalyzes phosphorolytic or arsenolytic citrulline cleavage; therefore, we examined if mitochondrial OCT facilitated AsV reduction in presence of glutathione. Isolated rat liver mitochondria were incubated with AsV, and AsIII formed was quantified. Glutathione-supplemented permeabilized or solubilized mitochondria reduced AsV. Citrulline (substrate for OCT-catalyzed arsenolysis) increased AsV reduction. The citrulline-stimulated AsV reduction was abolished by ornithine (OCT substrate inhibiting citrulline cleavage), phosphate (OCT substrate competing with AsV), and the OCT inhibitor norvaline or PALO, indicating that AsV reduction is coupled to OCT-catalyzed arsenolysis of citrulline. Corroborating this conclusion, purified bacterial OCT mediated AsV reduction in presence of citrulline and glutathione with similar responsiveness to these agents. In contrast, AsIII formation by intact mitochondria was unaffected by PALO and slightly stimulated by citrulline, ornithine, and norvaline, suggesting minimal role for OCT in AsV reduction in intact mitochondria. In addition to OCT, mitochondrial PNP can also mediate AsIII formation; however, its role in AsV reduction appears severely limited by purine nucleoside supply. Collectively, mitochondrial and bacterial OCT promote glutathione-dependent AsV reduction with coupled arsenolysis of citrulline, supporting the hypothesis that AsV reduction is mediated by phosphorolytic/arsenolytic enzymes. Nevertheless, because citrulline cleavage is disfavored physiologically, OCT may have little role in AsV reduction in vivo.

  6. The effect of chemical carcinogenesis on rat glutathione S-transferase P1 gene transcriptional regulation.

    Science.gov (United States)

    Liu, D; Liao, M; Zuo, J; Henner, W D; Fan, F

    2001-03-01

    To investigate mechanisms of rat glutathione S-transferase P1 gene (rGSTP1) expression regulation during chemical carcinogenesis. we studied enhancer elements located in the region between -2.5 kb to -2.2 kb. The region was upstream from the start site of transcription and was divided into two major fragments, GPEI and GPEII. The GPEII fragment was further divided into two smaller fragments, GPEII- I and GPEII-2. Using a luciferase reporter system, we identified a strong enhancer of GPEI and a weak enhancer of GPEII in HeLa and a rat hepatoma cell line CBRH79 19 cell. The enhancer of GPEII was located within the GPEII-I region. Chemical stimulation by glycidyl methatylate (GMA) and phorbol 12-o-tetradecanoate 13-acetate (TPA) analysis revealed that induction of rGSTP1 expression was mainly through GPEI. Although H2O2 could enhance GPEII enhancer activity, the enhancement is not mediated by the NF-kappaB factor that bound the NF-kappaB site in GPEII. Using electrophoretic mobility shift assays (EMSA) and the UV cross-linking assays, we found that HeLa and CBRH7919 cells had proteins that specifically bound GPEI core sequence and a 64 kDa protein that interacted with GPEII-1. The cells from normal rat liver did not express the binding proteins. Therefore, the trans-acting factors seem to be closely related to GPEI, GPEII enhancer activities and may play an important role in high expression of rGSTPI gene.

  7. Glutathione S-transferase expression and isoenzyme composition during cell differentiation of Caco-2 cells

    International Nuclear Information System (INIS)

    Scharmach, E.; Hessel, S.; Niemann, B.; Lampen, A.

    2009-01-01

    The human colon adenocarcinoma cell line Caco-2 is frequently used to study human intestinal metabolism and transport of xenobiotica. Previous studies have shown that both Caco-2 cells and human colon cells constitutively express the multigene family of detoxifying enzymes glutathione S-transferases (GSTs), particularly GST alpha and GST pi. GSTs may play a fundamental role in the molecular interplay between phase I, II enzymes and ABC-transporters. The gut fermentation product, butyrate, can modulate the potential for detoxification. The aim of this study was to investigate the basal expression of further cytosolic GSTs in Caco-2 cells during cell differentiation. In addition, a comparison was made with expression levels in MCF-7 and HepG2, two other cell types with barrier functions. Finally, the butyrate-mediated modulation of gene and protein expression was determined by real time PCR and western blot analysis. In Caco-2, gene and protein expression levels of GST alpha increased during cell differentiation. High levels of GSTO1 and GSTP1 were constantly expressed. No expression of GSTM5 and GSTT1 was detected. HepG2 expressed GSTO1 and MCF-7 GSTZ1 most intensively. No expression of GSTA5, GSTM5, or GSTP1 was detected in either cell. Incubation of Caco-2 cells with butyrate (5 mM) significantly induced GSTA1 and GSTM2 in proliferating Caco-2 cells. In differentiated cells, butyrate tended to increase GSTO1 and GSTP1. The results of this study show that a differentiation-dependent expression of GSTs in Caco-2 cells may reflect the in vivo situation and indicate the potential of butyrate to modify intestinal metabolism. GSTA1-A4 have been identified as good markers for cell differentiation. The Caco-2 cell line is a useful model for assessing the potential of food-related substances to modulate the GST expression pattern.

  8. Menadione stress in Saccharomyces cerevisiae strains deficient in the glutathione transferases.

    Science.gov (United States)

    Castro, F A V; Herdeiro, R S; Panek, A D; Eleutherio, E C A; Pereira, M D

    2007-02-01

    Using S. cerevisiae as a eukaryotic cell model we have analyzed the involvement of both glutathione transferase isoforms, Gtt1 and Gtt2, in constitutive resistance and adaptive response to menadione, a quinone which can exert its toxicity as redox cycling and/or electrophiles. The detoxification properties, of these enzymes, have also been analyzed by the appearance of S-conjugates in the media. Direct exposure to menadione (20 mM/60 min) showed to be lethal for cells deficient on both Gtt1 and Gtt2 isoforms. However, after pre-treatment with a low menadione concentration, cells deficient in Gtt2 displayed reduced ability to acquire tolerance when compared with the control and the Gtt1 deficient strains. Analyzing the toxic effects of menadione we observed that the gtt2 mutant showed no reduction in lipid peroxidation levels. Moreover, measuring the levels of intracellular oxidation during menadione stress we have shown that the increase of this oxidative stress parameter was due to the capacity menadione possesses in generating reactive oxygen species (ROS) and that both GSH and Gtt2 isoform were required to enhance ROS production. Furthermore, the efflux of the menadione-GSH conjugate, which is related with detoxification of xenobiotic pathways, was not detected in the gtt2 mutant. Taken together, these results suggest that acquisition of tolerance against stress generated by menadione and the process of detoxification through S-conjugates are dependent upon Gtt2 activity. This assessment was corroborated by the increase of GTT2 expression, and not of GTT1, after menadione treatment.

  9. Characterization and expression profiling of glutathione S-transferases in the diamondback moth, Plutella xylostella (L.).

    Science.gov (United States)

    You, Yanchun; Xie, Miao; Ren, Nana; Cheng, Xuemin; Li, Jianyu; Ma, Xiaoli; Zou, Minming; Vasseur, Liette; Gurr, Geoff M; You, Minsheng

    2015-03-05

    Glutathione S-transferases (GSTs) are multifunctional detoxification enzymes that play important roles in insects. The completion of several insect genome projects has enabled the identification and characterization of GST genes over recent years. This study presents a genome-wide investigation of the diamondback moth (DBM), Plutella xylostella, a species in which the GSTs are of special importance because this pest is highly resistant to many insecticides. A total of 22 putative cytosolic GSTs were identified from a published P. xylostella genome and grouped into 6 subclasses (with two unclassified). Delta, Epsilon and Omega GSTs were numerically superior with 5 genes for each of the subclasses. The resulting phylogenetic tree showed that the P. xylostella GSTs were all clustered into Lepidoptera-specific branches. Intron sites and phases as well as GSH binding sites were strongly conserved within each of the subclasses in the GSTs of P. xylostella. Transcriptome-, RNA-seq- and qRT-PCR-based analyses showed that the GST genes were developmental stage- and strain-specifically expressed. Most of the highly expressed genes in insecticide resistant strains were also predominantly expressed in the Malpighian tubules, midgut or epidermis. To date, this is the most comprehensive study on genome-wide identification, characterization and expression profiling of the GST family in P. xylostella. The diversified features and expression patterns of the GSTs are inferred to be associated with the capacity of this species to develop resistance to a wide range of pesticides and biological toxins. Our findings provide a base for functional research on specific GST genes, a better understanding of the evolution of insecticide resistance, and strategies for more sustainable management of the pest.

  10. Thermal- and urea-induced unfolding processes of glutathione S-transferase by molecular dynamics simulation.

    Science.gov (United States)

    Li, Jiahuang; Chen, Yuan; Yang, Jie; Hua, Zichun

    2015-05-01

    The Schistosoma juponicum 26 kDa glutathione S-transferase (sj26GST) consists of the N-terminal domain (N-domain), containing three alpha-helices (named H1-H3) and four anti-parallel beta-strands (S1-S4), and the C-terminal domain (C-domain), comprising five alpha-helices (named H4-H8). In present work, molecular dynamics simulations and fluorescence spectroscopic were used to gain insights into the unfolding process of sj26GST. The molecular dynamics simulations on sj26GST subunit both in water and in 8 M urea were carried out at 300 K, 400 K and 500 K, respectively. Spectroscopic measurements were employed to monitor structural changes. Molecular dynamics simulations of sj26GST subunit induced by urea and temperature showed that the initial unfolding step of sj26GST both in water and urea occurred on N-domain, involving the disruption of helices H2, H3 and strands S3 and S4, whereas H6 was the last region exposed to solution and was the last helix to unfold. Moreover, simulations analyses combining with fluorescence and circular dichroism spectra indicated that N-domain could not fold independent, suggesting that correct folding of N-domain depended on its interactions with C-domain. We further proposed that the folding of GSTs could begin with the hydrophobic collapse of C-domain whose H4, H5, H6 and H7 could move close to each other and form a hydrophobic core, especially H6 wrapped in the hydrophobic center and beginning spontaneous formation of the helix. S3, S4, H3, and H2 could form in the wake of the interaction between C-domain and N-domain. The paper can offer insights into the molecular mechanism of GSTs unfolding. © 2014 Wiley Periodicals, Inc.

  11. Glutathione S-transferase genotypes modify lung function decline in the general population: SAPALDIA cohort study

    Directory of Open Access Journals (Sweden)

    Ackermann-Liebrich Ursula

    2007-01-01

    Full Text Available Abstract Background Understanding the environmental and genetic risk factors of accelerated lung function decline in the general population is a first step in a prevention strategy against the worldwide increasing respiratory pathology of chronic obstructive pulmonary disease (COPD. Deficiency in antioxidative and detoxifying Glutathione S-transferase (GST gene has been associated with poorer lung function in children, smokers and patients with respiratory diseases. In the present study, we assessed whether low activity variants in GST genes are also associated with accelerated lung function decline in the general adult population. Methods We examined with multiple regression analysis the association of polymorphisms in GSTM1, GSTT1 and GSTP1 genes with annual decline in FEV1, FVC, and FEF25–75 during 11 years of follow-up in 4686 subjects of the prospective SAPALDIA cohort representative of the Swiss general population. Effect modification by smoking, gender, bronchial hyperresponisveness and age was studied. Results The associations of GST genotypes with FEV1, FVC, and FEF25–75 were comparable in direction, but most consistent for FEV1. GSTT1 homozygous gene deletion alone or in combination with GSTM1 homozygous gene deletion was associated with excess decline in FEV1 in men, but not women, irrespective of smoking status. The additional mean annual decline in FEV1 in men with GSTT1 and concurrent GSTM1 gene deletion was -8.3 ml/yr (95% confidence interval: -12.6 to -3.9 relative to men without these gene deletions. The GSTT1 effect on the FEV1 decline comparable to the observed difference in FEV1 decline between never and persistent smoking men. Effect modification by gender was statistically significant. Conclusion Our results suggest that genetic GSTT1 deficiency is a prevalent and strong determinant of accelerated lung function decline in the male general population.

  12. Farnesyl transferase inhibitors induce extended remissions in transgenic mice with mature B cell lymphomas

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    Refaeli Yosef

    2008-05-01

    Full Text Available Abstract Background We have used a mouse model based on overexpression of c-Myc in B cells genetically engineered to be self-reactive to test the hypothesis that farnesyl transferase inhibitors (FTIs can effectively treat mature B cell lymphomas. FTIs are undergoing clinical trials to treat both lymphoid and non-lymphoid malignancies and we wished to obtain evidence to support the inclusion of B cell lymphomas in future trials. Results We report that two FTIs, L-744,832 and SCH66336, blocked the growth of mature B cell lymphoma cells in vitro and in vivo. The FTI treatment affected the proliferation and survival of the transformed B cells to a greater extent than naïve B cells stimulated with antigen. In syngeneic mice transplanted with the transgenic lymphoma cells, L-744,832 treatment prevented the growth of the tumor cells and the morbidity associated with the resulting lymphoma progression. Tumors that arose from transplantation of the lymphoma cells regressed with as little as three days of treatment with L-744,832 or SCH66336. Treatment of these established lymphomas with L-744,832 for seven days led to long-term remission of the disease in approximately 25% of animals. Conclusion FTI treatment can block the proliferation and survival of self-reactive transformed B cells that overexpress Myc. In mice transplanted with mature B cell lymphomas, we found that FTI treatment led to regression of disease. FTIs warrant further consideration as therapeutic agents for mature B cell lymphomas and other lymphoid tumors.

  13. Enhanced tolerance and remediation of anthracene by transgenic tobacco plants expressing a fungal glutathione transferase gene

    Energy Technology Data Exchange (ETDEWEB)

    Dixit, Prachy; Mukherjee, Prasun K.; Sherkhane, Pramod D.; Kale, Sharad P. [Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Eapen, Susan, E-mail: eapenhome@yahoo.com [Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)

    2011-08-15

    Highlights: {yields} Transgenic plants expressing a TvGST gene were tested for tolerance, uptake and degradation of anthracene. {yields} Transgenic plants were more tolerant to anthracene and take up more anthracene from soil and solutions compared to control plants. {yields} Using in vitro T{sub 1} seedlings, we showed that anthracene-a three fused benzene ring compound was phytodegraded to naphthalene derivatives, having two benzene rings. {yields} This is the first time that a transgenic plant was shown to have the potential to phytodegrade anthracene. - Abstract: Plants can be used for remediation of polyaromatic hydrocarbons, which are known to be a major concern for human health. Metabolism of xenobiotic compounds in plants occurs in three phases and glutathione transferases (GST) mediate phase II of xenobiotic transformation. Plants, although have GSTs, they are not very efficient for degradation of exogenous recalcitrant xenobiotics including polyaromatic hydrocarbons. Hence, heterologous expression of efficient GSTs in plants may improve their remediation and degradation potential of xenobiotics. In the present study, we investigated the potential of transgenic tobacco plants expressing a Trichoderma virens GST for tolerance, remediation and degradation of anthracene-a recalcitrant polyaromatic hydrocarbon. Transgenic plants with fungal GST showed enhanced tolerance to anthracene compared to control plants. Remediation of {sup 14}C uniformly labeled anthracene from solutions and soil by transgenic tobacco plants was higher compared to wild-type plants. Transgenic plants (T{sub 0} and T{sub 1}) degraded anthracene to naphthalene derivatives, while no such degradation was observed in wild-type plants. The present work has shown that in planta expression of a fungal GST in tobacco imparted enhanced tolerance as well as higher remediation potential of anthracene compared to wild-type plants.

  14. Determination of the serine palmitoyl transferase inhibitor myriocin by electrospray and Q-trap mass spectrometry.

    Science.gov (United States)

    Campisi, Giuseppe Matteo; Signorelli, Paola; Rizzo, Jessica; Ghilardi, Claudio; Antognetti, Jacopo; Caretti, Anna; Lazarević, Jelena S; Strettoi, Enrica; Novelli, Elena; Ghidoni, Riccardo; Rubino, Federico Maria; Paroni, Rita

    2017-12-01

    Myriocin is a potent inhibitor of serine-palmitoyl-transferase, the first and rate-determining enzyme in the sphingolipids biosynthetic pathway. This study developed, validated and applied a LC-MS/MS method to measure myriocin in minute specimens of animal tissue. The chemical analog 14-OH-myriocin was used as the internal standard. The two molecules were extracted from the tissue homogenate by solid-phase extraction, separated by gradient reversed-phase liquid chromatography and measured by negative ion electrospray mass spectrometry in the triple quadrupole. Detection was accomplished by multiple reaction monitoring, employing the most representative transitions, 400@104 and 402@104 for myriocin and 14-OH-myriocin, respectively. The typical limit of detection and lower limit of quantitation of the optimized method were 0.9 pmol/mL (~0.016 pmol injected) and 2.3 pmol/mL, respectively, and the method was linear up to 250 pmol/mL range (r 2  = 0.9996). The intra- and between-day repeatability afforded a coefficient of variation ≤7.0%. Applications included quantification of myriocin in mouse lungs after 24 h from administration of ~4 nmol by intra-tracheal delivery. Measured levels ranged from 4.11 (median; 2.3-7.4 IQR, n = 4) to 11.7 (median; 7.6-22.7 interquartile range (IQR), n = 6) pmol/lung depending on the different formulations used. Myriocin was also measured in retinas of mice treated by intravitreal injection and ranged from 0.045 (less than the limit of detection) to 0.35 pmol/retina. Copyright © 2017 John Wiley & Sons, Ltd.

  15. Cloning of a glutathione S-transferase decreasing during differentiation of HL60 cell line

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jae Chul; Park, In Kyu; Lee, Kyu Bo; Sohn, Sang Kyun; Kim, Moo Kyu; Kim, Jung Chul [College of Medicine, Kyungpook National Univ., Taegu (Korea, Republic of)

    1999-06-01

    By sequencing the Expressed Sequence Tags of human dermal papilla cDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL60 cell line. K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Northern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusion expression system and the protein product was identified on SDS-PAGE. K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares 70% identity with that of rat glutathione S-transferase kappa 1 (rGSTK1). The transcripts were expressed inh a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in colorectal cancer and melanoma cell lines. Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

  16. Constitutive NADPH-dependent electron transferase activity of the Nox4 dehydrogenase domain.

    Science.gov (United States)

    Nisimoto, Yukio; Jackson, Heather M; Ogawa, Hisamitsu; Kawahara, Tsukasa; Lambeth, J David

    2010-03-23

    NADPH oxidase 4 (Nox4) is constitutively active, while Nox2 requires the cytosolic regulatory subunits p47(phox) and p67(phox) and activated Rac with activation by phorbol 12-myristate 13-acetate (PMA). This study was undertaken to identify the domain on Nox4 that confers constitutive activity. Lysates from Nox4-expressing cells exhibited constitutive NADPH- but not NADH-dependent hydrogen peroxide production with a K(m) for NADPH of 55 +/- 10 microM. The concentration of Nox4 in cell lysates was estimated using Western blotting and allowed calculation of a turnover of approximately 200 mol of H(2)O(2) min(-1) (mol of Nox4)(-1). A chimeric protein (Nox2/4) consisting of the Nox2 transmembrane (TM) domain and the Nox4 dehydrogenase (DH) domain showed H(2)O(2) production in the absence of cytosolic regulatory subunits. In contrast, chimera Nox4/2, consisting of the Nox4 TM and Nox2 DH domains, exhibited PMA-dependent activation that required coexpression of regulatory subunits. Nox DH domains from several Nox isoforms were purified and evaluated for their electron transferase activities. Nox1 DH, Nox2 DH, and Nox5 DH domains exhibited barely detectable activities toward artificial electron acceptors, while the Nox4 DH domain exhibited significant rates of reduction of cytochrome c (160 min(-1), largely superoxide dismutase-independent), ferricyanide (470 min(-1)), and other electron acceptors (artificial dyes and cytochrome b(5)). Rates were similar to those observed for H(2)O(2) production by the Nox4 holoenzyme in cell lysates. The activity required added FAD and was seen with NADPH but not NADH. These results indicate that the Nox4 DH domain exists in an intrinsically activated state and that electron transfer from NADPH to FAD is likely to be rate-limiting in the NADPH-dependent reduction of oxygen by holo-Nox4.

  17. Glutathione-S-transferase profiles in the emerald ash borer, Agrilus planipennis.

    Science.gov (United States)

    Rajarapu, Swapna Priya; Mittapalli, Omprakash

    2013-05-01

    The emerald ash borer, Agrilus planipennis Fairmaire is a recently discovered invasive insect pest of ash, Fraxinus spp. in North America. Glutathione-S-transferases (GST) are a multifunctional superfamily of enzymes which function in conjugating toxic compounds to less toxic and excretable forms. In this study, we report the molecular characterization and expression patterns of different classes of GST genes in different tissues and developmental stages plus their specific activity. Multiple sequence alignment of all six A. planipennis GSTs (ApGST-E1, ApGST-E2, ApGST-E3, ApGST-O1, ApGST-S1 and ApGST-μ1) revealed conserved features of insect GSTs and a phylogenetic analysis grouped the GSTs within the epsilon, sigma, omega and microsomal classes of GSTs. Real time quantitative PCR was used to study field collected samples. In larval tissues high mRNA levels for ApGST-E1, ApGST-E3 and ApGST-O1 were obtained in the midgut and Malpighian tubules. On the other hand, ApGST-E2 and ApGST-S1 showed high mRNA levels in fat body and ApGST-μ1 showed constitutive levels in all the tissues assayed. During development, mRNA levels for ApGST-E2 were observed to be the highest in feeding instars, ApGST-S1 in prepupal instars; while the others showed constitutive patterns in all the developmental stages examined. At the enzyme level, total GST activity was similar in all the tissues and developmental stages assayed. Results obtained suggest that A. planipennis is potentially primed with GST-driven detoxification to metabolize ash allelochemicals. To our knowledge this study represents the first report of GSTs in A. planipennis and also in the family of wood boring beetles. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. Enhanced tolerance and remediation of anthracene by transgenic tobacco plants expressing a fungal glutathione transferase gene

    International Nuclear Information System (INIS)

    Dixit, Prachy; Mukherjee, Prasun K.; Sherkhane, Pramod D.; Kale, Sharad P.; Eapen, Susan

    2011-01-01

    Highlights: → Transgenic plants expressing a TvGST gene were tested for tolerance, uptake and degradation of anthracene. → Transgenic plants were more tolerant to anthracene and take up more anthracene from soil and solutions compared to control plants. → Using in vitro T 1 seedlings, we showed that anthracene-a three fused benzene ring compound was phytodegraded to naphthalene derivatives, having two benzene rings. → This is the first time that a transgenic plant was shown to have the potential to phytodegrade anthracene. - Abstract: Plants can be used for remediation of polyaromatic hydrocarbons, which are known to be a major concern for human health. Metabolism of xenobiotic compounds in plants occurs in three phases and glutathione transferases (GST) mediate phase II of xenobiotic transformation. Plants, although have GSTs, they are not very efficient for degradation of exogenous recalcitrant xenobiotics including polyaromatic hydrocarbons. Hence, heterologous expression of efficient GSTs in plants may improve their remediation and degradation potential of xenobiotics. In the present study, we investigated the potential of transgenic tobacco plants expressing a Trichoderma virens GST for tolerance, remediation and degradation of anthracene-a recalcitrant polyaromatic hydrocarbon. Transgenic plants with fungal GST showed enhanced tolerance to anthracene compared to control plants. Remediation of 14 C uniformly labeled anthracene from solutions and soil by transgenic tobacco plants was higher compared to wild-type plants. Transgenic plants (T 0 and T 1 ) degraded anthracene to naphthalene derivatives, while no such degradation was observed in wild-type plants. The present work has shown that in planta expression of a fungal GST in tobacco imparted enhanced tolerance as well as higher remediation potential of anthracene compared to wild-type plants.

  19. Genetic polymorphism in three glutathione s-transferase genes and breast cancer risk

    Energy Technology Data Exchange (ETDEWEB)

    Woldegiorgis, S.; Ahmed, R.C.; Zhen, Y.; Erdmann, C.A.; Russell, M.L.; Goth-Goldstein, R.

    2002-04-01

    The role of the glutathione S-transferase (GST) enzyme family is to detoxify environmental toxins and carcinogens and to protect organisms from their adverse effects, including cancer. The genes GSTM1, GSTP1, and GSTT1 code for three GSTs involved in the detoxification of carcinogens, such as polycyclic aromatic hydrocarbons (PAHs) and benzene. In humans, GSTM1 is deleted in about 50% of the population, GSTT1 is absent in about 20%, whereas the GSTP1 gene has a single base polymorphism resulting in an enzyme with reduced activity. Epidemiological studies indicate that GST polymorphisms increase the level of carcinogen-induced DNA damage and several studies have found a correlation of polymorphisms in one of the GST genes and an increased risk for certain cancers. We examined the role of polymorphisms in genes coding for these three GST enzymes in breast cancer. A breast tissue collection consisting of specimens of breast cancer patients and non-cancer controls was analyzed by polymerase chain reaction (PCR) for the presence or absence of the GSTM1 and GSTT1 genes and for GSTP1 single base polymorphism by PCR/RFLP. We found that GSTM1 and GSTT1 deletions occurred more frequently in cases than in controls, and GSTP1 polymorphism was more frequent in controls. The effective detoxifier (putative low-risk) genotype (defined as presence of both GSTM1 and GSTT1 genes and GSTP1 wild type) was less frequent in cases than controls (16% vs. 23%, respectively). The poor detoxifier (putative high-risk) genotype was more frequent in cases than controls. However, the sample size of this study was too small to provide conclusive results.

  20. OGT (O-GlcNAc Transferase) Selectively Modifies Multiple Residues Unique to Lamin A.

    Science.gov (United States)

    Simon, Dan N; Wriston, Amanda; Fan, Qiong; Shabanowitz, Jeffrey; Florwick, Alyssa; Dharmaraj, Tejas; Peterson, Sherket B; Gruenbaum, Yosef; Carlson, Cathrine R; Grønning-Wang, Line M; Hunt, Donald F; Wilson, Katherine L

    2018-05-17

    The LMNA gene encodes lamins A and C with key roles in nuclear structure, signaling, gene regulation, and genome integrity. Mutations in LMNA cause over 12 diseases ('laminopathies'). Lamins A and C are identical for their first 566 residues. However, they form separate filaments in vivo, with apparently distinct roles. We report that lamin A is β- O -linked N -acetylglucosamine- (O -GlcNAc)-modified in human hepatoma (Huh7) cells and in mouse liver. In vitro assays with purified O -GlcNAc transferase (OGT) enzyme showed robust O -GlcNAcylation of recombinant mature lamin A tails (residues 385⁻646), with no detectable modification of lamin B1, lamin C, or 'progerin' (Δ50) tails. Using mass spectrometry, we identified 11 O -GlcNAc sites in a 'sweet spot' unique to lamin A, with up to seven sugars per peptide. Most sites were unpredicted by current algorithms. Double-mutant (S612A/T643A) lamin A tails were still robustly O -GlcNAc-modified at seven sites. By contrast, O -GlcNAcylation was undetectable on tails bearing deletion Δ50, which causes Hutchinson⁻Gilford progeria syndrome, and greatly reduced by deletion Δ35. We conclude that residues deleted in progeria are required for substrate recognition and/or modification by OGT in vitro. Interestingly, deletion Δ35, which does not remove the majority of identified O -GlcNAc sites, does remove potential OGT-association motifs (lamin A residues 622⁻625 and 639⁻645) homologous to that in mouse Tet1. These biochemical results are significant because they identify a novel molecular pathway that may profoundly influence lamin A function. The hypothesis that lamin A is selectively regulated by OGT warrants future testing in vivo, along with two predictions: genetic variants may contribute to disease by perturbing OGT-dependent regulation, and nutrient or other stresses might cause OGT to misregulate wildtype lamin A.

  1. O-GlcNAc transferase regulates transcriptional activity of human Oct4.

    Science.gov (United States)

    Constable, Sandii; Lim, Jae-Min; Vaidyanathan, Krithika; Wells, Lance

    2017-10-01

    O-linked β-N-acetylglucosamine (O-GlcNAc) is a single sugar modification found on many different classes of nuclear and cytoplasmic proteins. Addition of this modification, by the enzyme O-linked N-acetylglucosamine transferase (OGT), is dynamic and inducible. One major class of proteins modified by O-GlcNAc is transcription factors. O-GlcNAc regulates transcription factor properties through a variety of different mechanisms including localization, stability and transcriptional activation. Maintenance of embryonic stem (ES) cell pluripotency requires tight regulation of several key transcription factors, many of which are modified by O-GlcNAc. Octamer-binding protein 4 (Oct4) is one of the key transcription factors required for pluripotency of ES cells and more recently, the generation of induced pluripotent stem (iPS) cells. The action of Oct4 is modulated by the addition of several post-translational modifications, including O-GlcNAc. Previous studies in mice found a single site of O-GlcNAc addition responsible for transcriptional regulation. This study was designed to determine if this mechanism is conserved in humans. We mapped 10 novel sites of O-GlcNAc attachment on human Oct4, and confirmed a role for OGT in transcriptional activation of Oct4 at a site distinct from that found in mouse that allows distinction between different Oct4 target promoters. Additionally, we uncovered a potential new role for OGT that does not include its catalytic function. These results confirm that human Oct4 activity is being regulated by OGT by a mechanism that is distinct from mouse Oct4. © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Glutathione S-transferase expression and isoenzyme composition during cell differentiation of Caco-2 cells.

    Science.gov (United States)

    Scharmach, E; Hessel, S; Niemann, B; Lampen, A

    2009-11-30

    The human colon adenocarcinoma cell line Caco-2 is frequently used to study human intestinal metabolism and transport of xenobiotica. Previous studies have shown that both Caco-2 cells and human colon cells constitutively express the multigene family of detoxifying enzymes glutathione S-transferases (GSTs), particularly GST alpha and GST pi. GSTs may play a fundamental role in the molecular interplay between phase I, II enzymes and ABC-transporters. The gut fermentation product, butyrate, can modulate the potential for detoxification. The aim of this study was to investigate the basal expression of further cytosolic GSTs in Caco-2 cells during cell differentiation. In addition, a comparison was made with expression levels in MCF-7 and HepG2, two other cell types with barrier functions. Finally, the butyrate-mediated modulation of gene and protein expression was determined by real time PCR and western blot analysis. In Caco-2, gene and protein expression levels of GST alpha increased during cell differentiation. High levels of GSTO1 and GSTP1 were constantly expressed. No expression of GSTM5 and GSTT1 was detected. HepG2 expressed GSTO1 and MCF-7 GSTZ1 most intensively. No expression of GSTA5, GSTM5, or GSTP1 was detected in either cell. Incubation of Caco-2 cells with butyrate (5 mM) significantly induced GSTA1 and GSTM2 in proliferating Caco-2 cells. In differentiated cells, butyrate tended to increase GSTO1 and GSTP1. The results of this study show that a differentiation-dependent expression of GSTs in Caco-2 cells may reflect the in vivo situation and indicate the potential of butyrate to modify intestinal metabolism. GSTA1-A4 have been identified as good markers for cell differentiation. The Caco-2 cell line is a useful model for assessing the potential of food-related substances to modulate the GST expression pattern.

  3. Primary and secondary structural analyses of glutathione S-transferase pi from human placenta.

    Science.gov (United States)

    Ahmad, H; Wilson, D E; Fritz, R R; Singh, S V; Medh, R D; Nagle, G T; Awasthi, Y C; Kurosky, A

    1990-05-01

    The primary structure of glutathione S-transferase (GST) pi from a single human placenta was determined. The structure was established by chemical characterization of tryptic and cyanogen bromide peptides as well as automated sequence analysis of the intact enzyme. The structural analysis indicated that the protein is comprised of 209 amino acid residues and gave no evidence of post-translational modifications. The amino acid sequence differed from that of the deduced amino acid sequence determined by nucleotide sequence analysis of a cDNA clone (Kano, T., Sakai, M., and Muramatsu, M., 1987, Cancer Res. 47, 5626-5630) at position 104 which contained both valine and isoleucine whereas the deduced sequence from nucleotide sequence analysis identified only isoleucine at this position. These results demonstrated that in the one individual placenta studied at least two GST pi genes are coexpressed, probably as a result of allelomorphism. Computer assisted consensus sequence evaluation identified a hydrophobic region in GST pi (residues 155-181) that was predicted to be either a buried transmembrane helical region or a signal sequence region. The significance of this hydrophobic region was interpreted in relation to the mode of action of the enzyme especially in regard to the potential involvement of a histidine in the active site mechanism. A comparison of the chemical similarity of five known human GST complete enzyme structures, one of pi, one of mu, two of alpha, and one microsomal, gave evidence that all five enzymes have evolved by a divergent evolutionary process after gene duplication, with the microsomal enzyme representing the most divergent form.

  4. Effects of toasting blue lupins, soybeans or barley as supplement for high-yielding organic dairy cows fed grass-clover silage ad libitum

    DEFF Research Database (Denmark)

    Mogensen, Lisbeth; Lund, Peter; Kristensen, Troels

    2008-01-01

    . The supplement under investigation was: lupins in experiment 1, barley in experiment 2 and soybeans in experiment 3. The same forage mixture of grass-clover silage (84% of DM), grass pellets (11% of DM) and straw (5% of DM) was fed ad libitum in all the experiments. Toasting decreased effective rumen protein...... degradability determined in situ for all three supplements. Compared to untreated lupins toasting of lupins tended (P = 0.10) to increase milk yield, whereas toasting of soybeans did not affect milk yield. Toasting of lupins decreased (P = 0.03) milk protein content (32.2 versus 32.7 g/kg), whereas toasting...... of soybeans did not affect milk protein content. ECM yield was significantly higher (P = 0.002) for cows fed toasted soybeans than for cows fed untreated soybeans (28.1 versus 26.4 kg ECM) whereas there was no significant effect on ECM yield from toasting lupins or barley. It can be concluded...

  5. Lattice Calculation of D- and B-meson Semileptonic Decays, using the Clover Action at beta=6.0 on APE

    CERN Document Server

    Allton, C R; Lubicz, V; Martinelli, G; Rapuano, F; Stella, N; Vladikas, A; Bartoloni, A; Battista, C; Cabasino, S; Cabibbo, Nicola; Panizzi, E; Paolucci, P S; Sarno, R; Todesco, G M; Torelli, M; Vicini, P

    1995-01-01

    We present the results of a high statistics lattice calculation of hadronic form factors relevant for $D-$ and $B-$meson semi-leptonic decays into light pseudoscalar and vector mesons. The results have been obtained by averaging over 170 gauge field configurations, generated in the quenched approximation, at $\\beta=6.0$, on a $18^3 \\times 64$ lattice, using the $O(a)$-improved SW-Clover action.From the study of the matrix element $$, we obtain $f_+ (0)=0.78\\pm 0.08$ and from the matrix element $$ we obtain $V(0)=1.08\\pm 0.22$, $A_1(0)=0.67\\pm 0.11$ and $A_2(0)=0.49\\pm 0.34$. We also obtain the ratios $V(0)/A_1(0)=1.6\\pm 0.3$ and $A_2(0)/A_1(0)= 0.7\\pm 0.4$. Our predictions for the different form factors are in good agreement with the experimental data, although, in the case of $A_2(0)$, the errors are still too large to draw any firm conclusion. With the help of the Heavy Quark Effective Theory (HQET) we have also extrapolated the lattice results to $B$-meson decays. The form factors follow a behaviour compat...

  6. Evalution of the healthiness of winter wheat cultivated in conventional tillage, direct sowing and direct sowing with underplant crop of white clover

    Directory of Open Access Journals (Sweden)

    Ewa Moszczyńska

    2012-12-01

    Full Text Available Research of the healthiness of winter wheat depending on the soil tillage system and rate of nitrogen fertilization were carried out in 1998-2001. The largest threat to the healthiness of plants was tan spot, which was caused by Pyrenophora tritici-repentis, especially in cropping season 1999/2000. The soil tillage system diversified the intensification of occurence of this pathogen, only in two last years of research. The most infected by P. tritici-i was wheat, which was cultivated in the direct sowing. Application of underplant crop of white clover in the direct sowing contributed to the improvement of the plants healthiness. The highest rate of nitrogen fertilization (120 kg N.ha-1 in the highest degree favoured the damage of wheat by P. tritici-repentis, but only in two first years of research. The second pathogen Blumeria graminis, which caused powdery mildew of cereals, occured in small amount and didn't have any influence on the healthiness of winter wheat.

  7. Inclusion of sunflower seed and wheat dried distillers' grains with solubles in a red clover silage-based diet enhances steers performance, meat quality and fatty acid profiles.

    Science.gov (United States)

    Mapiye, C; Aalhus, J L; Turner, T D; Vahmani, P; Baron, V S; McAllister, T A; Block, H C; Uttaro, B; Dugan, M E R

    2014-12-01

    The current study compared beef production, quality and fatty acid (FA) profiles of yearling steers fed a control diet containing 70 : 30 red clover silage (RCS) : barley-based concentrate, a diet containing 11% sunflower seed (SS) substituted for barley, and diets containing SS with 15% or 30% wheat dried distillers' grain with solubles (DDGS). Additions of DDGS were balanced by reductions in RCS and SS to maintain crude fat levels in diets. A total of two pens of eight animals were fed per diet for an average period of 208 days. Relative to the control diet, feeding the SS diet increased (Pproducts (i.e. atypical dienes) with the first double bond at carbon 8 or 9 from the carboxyl end, conjugated linoleic acid isomers with the first double bond from carbon 7 to 10 from the carboxyl end, t-18:1 isomers, and reduced (Pmeat tenderness. However, in general feeding DGGS-15 or DDGS-30 diets did not change FA proportions relative to feeding the SS diet. Overall, adding SS to a RCS-based diet enhanced muscle proportions of 18:2n-6 biohydrogenation products, and further substitutions of DDGS in the diet improved beef production, and quality while maintaining proportions of potentially functional bioactive FA including vaccenic and rumenic acids.

  8. Selection of the Optimal Herbal Compositions of Red Clover and Pomegranate According to Their Protective Effect against Climacteric Symptoms in Ovariectomized Mice

    Directory of Open Access Journals (Sweden)

    Su Jin Kang

    2016-07-01

    Full Text Available This study aimed to ascertain the optimal range of red clover dry extracts (RC and dried pomegranate concentrate powder (PCP to induce anti-climacteric effects. Thus, the dose ranges showing protective effect of mixed formulae consisting of RC and PCP were examined in ovariectomized mice. At 28 days after bilateral ovariectomy (OVX, mixed herbal compositions (RC:PCP = 1:1, 1:2, 1:4, 1:8, 2:1, 4:1, and 8:1 were administered orally, at 120 mg/kg once daily for 84 days. We evaluated that RC and PCP mixture attenuate OVX-caused obesity, hyperlipidemia, hepatic steatosis, and osteoporosis. Compared to OVX-induced control mice, body weight and abdominal fat weight in OVX-induced mice were significantly decreased, concomitantly with increase of uterus weight by RC:PCP mixture. Additionally, significant increases in serum estradiol levels were observed in all RC:PCP-treated mice. RC:PCP mixture also showed protective effect against OVX-induced hyperlipidemia, hepatic steatosis. Total body and femur mean bone mineral density (BMD, osteocalcin, bALP contents were effectively increased by RC:PCP mixture. Taken together, RC:PCP mixture (2:1, 1:1, and 4:1 has remarkable protective effects against the changes induced by OVX. In particular, RC:PCP mixture (2:1 shows the strongest effect and may be considered as a potential protective agent against climacteric symptoms.

  9. Mitigating N2O emissions from clover residues by 3,4-dimethylpyrazole phosphate (DMPP) without adverse effects on the earthworm Lumbricus terrestris

    DEFF Research Database (Denmark)

    Kong, Xianwang; Duan, YunFeng; Schramm, Andreas

    2017-01-01

    in a 28-d mesocosm experiment, where DMPP spraying prior to tillage was simulated. Above-ground parts of 15N-labelled clover residues were treated with DMPP and either placed at 10 cm depth to simulate ploughing (PL), or mixed with soil at 0–10 cm depth to simulate rotovation (RO). Earthworms (Lumbricus...... terrestris) were introduced to study their role in residue decomposition and N2O emissions. Fluxes and isotopic composition of N2O were determined with dynamic chambers using laser spectroscopy. A gradual increase in 15N-enrichment of N2O indicated that denitrification was the main source. DMPP reduced...... cumulative N2O emissions in PL from 241 to 146 mg N m−2; the reduction in RO was smaller, from 103 to 94 mg N m−2, and not significant, possibly due to higher oxygen and soil NO3− availability. After 28 d incubation, on average > 90% of the earthworms were recovered, and in vivo N2O production from L...

  10. Discrimination of Clover and Citrus Honeys from Egypt According to Floral Type Using Easily Assessable Physicochemical Parameters and Discriminant Analysis: An External Validation of the Chemometric Approach

    Directory of Open Access Journals (Sweden)

    Ioannis K. Karabagias

    2018-05-01

    Full Text Available Twenty-two honey samples, namely clover and citrus honeys, were collected from the greater Cairo area during the harvesting year 2014–2015. The main purpose of the present study was to characterize the aforementioned honey types and to investigate whether the use of easily assessable physicochemical parameters, including color attributes in combination with chemometrics, could differentiate honey floral origin. Parameters taken into account were: pH, electrical conductivity, ash, free acidity, lactonic acidity, total acidity, moisture content, total sugars (degrees Brix-°Bx, total dissolved solids and their ratio to total acidity, salinity, CIELAB color parameters, along with browning index values. Results showed that all honey samples analyzed met the European quality standards set for honey and had variations in the aforementioned physicochemical parameters depending on floral origin. Application of linear discriminant analysis showed that eight physicochemical parameters, including color, could classify Egyptian honeys according to floral origin (p < 0.05. Correct classification rate was 95.5% using the original method and 90.9% using the cross validation method. The discriminatory ability of the developed model was further validated using unknown honey samples. The overall correct classification rate was not affected. Specific physicochemical parameter analysis in combination with chemometrics has the potential to enhance the differences in floral honeys produced in a given geographical zone.

  11. Forage yield and nutritive value of Elephant grass, Italian ryegrass and spontaneous growing species mixed with forage peanut or red clover

    Directory of Open Access Journals (Sweden)

    Michelle Schalemberg Diehl

    2014-10-01

    Full Text Available The objective of this research was to evaluate of three grazing systems (GS with elephant grass (EG, Italian ryegrass (IR + spontaneous growing species (SGS; EG + IR + SGS + forage peanut (FP; and EG + IR + SGS + red clover (RC, during the winter and summer periods in rotational grazing with dairy cattle. Experimental design was completely randomized with three treatments, two replicates with repeated measures. Lactating Holstein cows receiving 1% BW-daily feed supplement with concentrate were used in the evaluation. Eight grazing cycles were performed during the experimental period. The values of pre forage mass and stocking rate were 2.52, 2.60 and 2.99 t ha-1 and 2.64, 2.77 and 3.14 animal unit ha-1, respectively for GS. Samples of forage were collected by hand-plucking technique to analyze the crude protein (CP, neutral detergent fiber (NDF, in situ dry matter digestibility (ISDMD, in situ organic matter digestibility (ISOMD of forage present between rows of elephant grass, in the rows of elephant grass and the legumes. Higher value of CP, ISOMD and lower of NDF were observed for the grazing systems mixed with legumes forage.

  12. In vivo induction of phase II detoxifying enzymes, glutathione transferase and quinone reductase by citrus triterpenoids

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    Ahmad Hassan

    2010-09-01

    Full Text Available Abstract Background Several cell culture and animal studies demonstrated that citrus bioactive compounds have protective effects against certain types of cancer. Among several classes of citrus bioactive compounds, limonoids were reported to prevent different types of cancer. Furthermore, the structures of citrus limonoids were reported to influence the activity of phase II detoxifying enzymes. The purpose of the study was to evaluate how variations in the structures of citrus limonoids (namely nomilin, deacetyl nomilin, and isoobacunoic acid and a mixture of limonoids would influence phase II enzyme activity in excised tissues from a mouse model. Methods In the current study, defatted sour orange seed powder was extracted with ethyl acetate and subjected to silica gel chromatography. The HPLC, NMR and mass spectra were used to elucidate the purity and structure of compounds. Female A/J mice were treated with three limonoids and a mixture in order to evaluate their effect on phase II enzymes in four different tissues. Assays for glutathione S-transferase and NAD(PH: quinone reductase (QR were used to evaluate induction of phase II enzymatic activity. Results The highest induction of GST against 1-chloro-2,4-dinitrobenzene (CDNB was observed in stomach (whole, 58% by nomilin, followed by 25% isoobacunoic acid and 19% deacetyl nomilin. Deacetyl nomilin in intestine (small as well as liver significantly reduced GST activity against CDNB. Additionally isoobacunoic acid and the limonoid mixture in liver demonstrated a significant reduction of GST activity against CDNB. Nomilin significantly induced GST activity against 4-nitroquinoline 1-oxide (4NQO, intestine (280% and stomach (75% while deacetyl nomilin showed significant induction only in intestine (73%. Induction of GST activity was also observed in intestine (93% and stomach (45% treated with the limonoid mixture. Finally, a significant induction of NAD(PH: quinone reductase (QR activity was

  13. Glutathione S-transferase activity in follicular fluid from women undergoing ovarian stimulation: role in maturation.

    Science.gov (United States)

    Meijide, Susana; Hernández, M Luisa; Navarro, Rosaura; Larreategui, Zaloa; Ferrando, Marcos; Ruiz-Sanz, José Ignacio; Ruiz-Larrea, M Begoña

    2014-10-01

    Female infertility involves an emotional impact for the woman, often leading to a state of anxiety and low self-esteem. The assisted reproduction techniques (ART) are used to overcome the problem of infertility. In a first step of the in vitro fertilization therapy women are subjected to an ovarian stimulation protocol to obtain mature oocytes, which will result in competent oocytes necessary for fertilization to occur. Ovarian stimulation, however, subjects the women to a high physical and psychological stress, thus being essential to improve ART and to find biomarkers of dysfunction and fertility. GSH is an important antioxidant, and is also used in detoxification reactions, catalysed by glutathione S-transferases (GST). In the present work, we have investigated the involvement of GST in follicular maturation. Patients with fertility problems and oocyte donors were recruited for the study. From each woman follicles at two stages of maturation were extracted at the preovulatory stage. Follicular fluid was separated from the oocyte by centrifugation and used as the enzyme source. GST activity was determined based on its conjugation with 3,4-dichloronitrobenzene and the assay was adapted to a 96-well microplate reader. The absorbance was represented against the incubation time and the curves were adjusted to linearity (R(2)>0.990). Results showed that in both donors and patients GST activity was significantly lower in mature oocytes compared to small ones. These results suggest that GST may play a role in the follicle maturation by detoxifying xenobiotics, thus contributing to the normal development of the oocyte. Supported by FIS/FEDER (PI11/02559), Gobierno Vasco (Dep. Educación, Universiades e Investigación, IT687-13), and UPV/EHU (CLUMBER UFI11/20 and PES13/58). The work was approved by the Ethics Committee of the UPV/EHU (CEISH/96/2011/RUIZLARREA), and performed according to the UPV/EHU and IVI-Bilbao agreement (Ref. 2012/01). Copyright © 2014. Published by

  14. Clonorchis sinensis omega-class glutathione transferases are reliable biomarkers for serodiagnosis of clonorchiasis and opisthorchiasis.

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    Kim, J-G; Ahn, C-S; Sripa, B; Eom, K S; Kang, I; Sohn, W-M; Nawa, Y; Kong, Y

    2018-04-09

    To determine the potential for immunodiagnostic application of two recombinant forms of Clonorchis sinensis omega-class glutathione transferases (rCsGSTo1 and rCsGSTo2) against human small liver-fluke C. sinensis and Opisthorchis viverrini infections. Specific antibody levels against rCsGSTo1 and rCsGSTo2 in patients' sera of egg-positive opisthorchiasis (n = 87) and clonorchiasis (n = 120), as well as those in sera from patients with other helminthic infections (n = 252) and healthy controls (n = 40) were retrospectively analysed by ELISA. We observed highly positive correlation coefficients between specific antibody levels against rCsGSTo1 and rCsGSTo2 and egg counts per gramme of faeces (EPG) of patients with opisthorchiasis (n = 87; r = 0.88 for rCsGSTo1 and r = 0.90 for rCsGSTo2). Sera from opisthorchiasis patients whose EPG counts >100 (n = 43) revealed high antibody titres against both antigens. Patients' sera with low EPG counts (<100, n = 44) also exhibited reliable sensitivities of 93.2% and 97.7% for rCsGSTo1 and rCsGSTo2, respectively. Sera from clonorchiasis patients showed sensitivities of 90% (108/120 samples) and 89.2% (107/120 sera) for rCsGSTo1 and rCsGSTo2. Overall diagnostic sensitivities for liver-fluke infections were 92.3% for rCsGSTo1 (191/207 samples) and 93.2% for rCsGSTo2 (193/207 samples). Specificities were 89.7% (rCsGSTo1) and 97.6% (rCsGSTo2). Detection of specific antibody levels against rCsGSTo1 or rCsGSTo2 might be promising for the serodiagnosis of patients infected with these two phylogenetically close carcinogenic liver-flukes. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  15. SERUM GAMMA-GLUTAMYL TRANSFERASE AS A BIOMARKER OF TYPE-2 DM AMONG CIGARETTE SMOKERS

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    K. Suganthy

    2017-03-01

    Full Text Available BACKGROUND Smoking is one of the most common addictions of modern times and needs to be studied in a community as a public health issue. Also, smoking is a modifiable risk factor for type-2 DM. The smoking-related diseases share common pathophysiologies of imbalance of systemic oxidants and antioxidant status, increased inflammatory reactions, insulin resistance and dyslipidaemia. Biochemical assay of serum Gamma-Glutamyl Transferase (GGT activity is a low cost and highly sensitive laboratory test. Studies have indicated GGT is moderately elevated before the onset of other traditional risk factors for type-2 DM. So, among hepatic markers, the baseline GGT analysis can be an early risk marker of type 2 diabetes in cigarette smokers has to be studied. MATERIALS AND METHODS This is a case-control study on male cigarette smokers. 57 smokers were studied clinically and biochemically for plasma insulin, glucose and liver enzymes including GGT using standard biochemical methods and compared with 42 age and sex matched non-smokers as controls. RESULTS The mean serum GGT in smokers (25.45 ± 10.8 was increased compared to non-smokers (18.8 ± 5.8. Smokers GGT (r=0.396 and HOMA-IR (r=0.352 showed significant positive association with duration of smoking (p24 IU/L. Regression analysis showed none of the diabetic risk factors were observed to be dependent on GGT including other liver enzymes. Regression analysis showed GGT is not an independent risk factor for DM. Although, the mean fasting blood glucose (91.4 ± 21.3, BMI (26.1 ± 9.3 and HOMA-IR (7.3 ± 2.3 was increased among cigarette smokers with GGT >24 IU/L. CONCLUSION The baseline GGT assay in cigarette smokers might be associated with the proinflammatory status or be a marker of oxidative stress of smoke toxins. Smokers with baseline GGT >24 IU/L develop insulin resistance should be investigated in future longitudinal studies for prediabetes to consider cigarette smoking as an important modifiable

  16. Effects of imidacloprid on detoxifying enzyme glutathione S-transferase on Folsomia candida (Collembola).

    Science.gov (United States)

    Sillapawattana, Panwad; Schäffer, Andreas

    2017-04-01

    Chemical analyses of the environment can document contamination by various xenobiotics, but it is also important to understand the effect of pollutants on living organisms. Thus, in the present work, we investigated the effect of the pesticide imidacloprid on the detoxifying enzyme glutathione S-transferase (GST) from Folsomia candida (Collembola), a standard test organism for estimating the effects of pesticides and environmental pollutants on non-target soil arthropods. Test animals were treated with different concentrations of imidacloprid for 48 h. Changes in steady-state levels of GST messenger RNA (mRNA) and GST enzyme activity were investigated. Extracted proteins were separated according to their sizes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the resolved protein bands were detected by silver staining. The size of the glutathione (GSH) pool in Collembola was also determined. A predicted protein sequence of putative GSTs was identified with animals from control group. A 3-fold up-regulation of GST steady-state mRNA levels was detected in the samples treated with 5.0 mg L -1 imidacloprid compared to the control, while a 2.5- and 2.0- fold up-regulation was found in organisms treated with 2.5 and 7.5 mg L -1 imidacloprid, respectively. GST activity increased with increasing imidacloprid amounts from an initial activity of 0.11 μmol min -1  mg -1 protein in the control group up to 0.25 μmol min -1  mg -1 protein in the sample treated with the 5.0 mg L -1 of pesticide. By contrast, the total amount of GSH decreased with increasing imidacloprid concentration. The results suggest that the alteration of GST activity, steady-state level of GST mRNA, and GSH level may be involved in the response of F. candida to the exposure of imidacloprid and can be used as biomarkers to monitor the toxic effects of imidacloprid and other environmental pollutants on Collembola.

  17. O-linked-N-acetylglucosamine modification of mammalian Notch receptors by an atypical O-GlcNAc transferase Eogt1

    Energy Technology Data Exchange (ETDEWEB)

    Sakaidani, Yuta [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Ichiyanagi, Naoki [Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan); Saito, Chika; Nomura, Tomoko [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Ito, Makiko [Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan); Nishio, Yosuke [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Nadano, Daita; Matsuda, Tsukasa [Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan); Furukawa, Koichi [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Okajima, Tetsuya, E-mail: tokajima@med.nagoya-u.ac.jp [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan)

    2012-03-02

    Highlights: Black-Right-Pointing-Pointer We characterized A130022J15Rik (Eogt1)-a mouse gene homologous to Drosophila Eogt. Black-Right-Pointing-Pointer Eogt1 encodes EGF domain O-GlcNAc transferase. Black-Right-Pointing-Pointer Expression of Eogt1 in Drosophila rescued the cell-adhesion defect in the Eogt mutant. Black-Right-Pointing-Pointer O-GlcNAcylation reaction in the secretory pathway is conserved through evolution. -- Abstract: O-linked-{beta}-N-acetylglucosamine (O-GlcNAc) modification is a unique cytoplasmic and nuclear protein modification that is common in nearly all eukaryotes, including filamentous fungi, plants, and animals. We had recently reported that epidermal growth factor (EGF) repeats of Notch and Dumpy are O-GlcNAcylated by an atypical O-GlcNAc transferase, EOGT, in Drosophila. However, no study has yet shown whether O-GlcNAcylation of extracellular proteins is limited to insects such as Drosophila or whether it occurs in other organisms, including mammals. Here, we report the characterization of A130022J15Rik, a mouse gene homolog of Drosophila Eogt (Eogt 1). Enzymatic analysis revealed that Eogt1 has a substrate specificity similar to that of Drosophila EOGT, wherein the Thr residue located between the fifth and sixth conserved cysteines of the folded EGF-like domains is modified. This observation is supported by the fact that the expression of Eogt1 in Drosophila rescued the cell-adhesion defect caused by Eogt downregulation. In HEK293T cells, Eogt1 expression promoted modification of Notch1 EGF repeats by O-GlcNAc, which was further modified, at least in part, by galactose to generate a novel O-linked-N-acetyllactosamine structure. These results suggest that Eogt1 encodes EGF domain O-GlcNAc transferase and that O-GlcNAcylation reaction in the secretory pathway is a fundamental biochemical process conserved through evolution.

  18. O-linked-N-acetylglucosamine modification of mammalian Notch receptors by an atypical O-GlcNAc transferase Eogt1

    International Nuclear Information System (INIS)

    Sakaidani, Yuta; Ichiyanagi, Naoki; Saito, Chika; Nomura, Tomoko; Ito, Makiko; Nishio, Yosuke; Nadano, Daita; Matsuda, Tsukasa; Furukawa, Koichi; Okajima, Tetsuya

    2012-01-01

    Highlights: ► We characterized A130022J15Rik (Eogt1)—a mouse gene homologous to Drosophila Eogt. ► Eogt1 encodes EGF domain O-GlcNAc transferase. ► Expression of Eogt1 in Drosophila rescued the cell-adhesion defect in the Eogt mutant. ► O-GlcNAcylation reaction in the secretory pathway is conserved through evolution. -- Abstract: O-linked-β-N-acetylglucosamine (O-GlcNAc) modification is a unique cytoplasmic and nuclear protein modification that is common in nearly all eukaryotes, including filamentous fungi, plants, and animals. We had recently reported that epidermal growth factor (EGF) repeats of Notch and Dumpy are O-GlcNAcylated by an atypical O-GlcNAc transferase, EOGT, in Drosophila. However, no study has yet shown whether O-GlcNAcylation of extracellular proteins is limited to insects such as Drosophila or whether it occurs in other organisms, including mammals. Here, we report the characterization of A130022J15Rik, a mouse gene homolog of Drosophila Eogt (Eogt 1). Enzymatic analysis revealed that Eogt1 has a substrate specificity similar to that of Drosophila EOGT, wherein the Thr residue located between the fifth and sixth conserved cysteines of the folded EGF-like domains is modified. This observation is supported by the fact that the expression of Eogt1 in Drosophila rescued the cell-adhesion defect caused by Eogt downregulation. In HEK293T cells, Eogt1 expression promoted modification of Notch1 EGF repeats by O-GlcNAc, which was further modified, at least in part, by galactose to generate a novel O-linked-N-acetyllactosamine structure. These results suggest that Eogt1 encodes EGF domain O-GlcNAc transferase and that O-GlcNAcylation reaction in the secretory pathway is a fundamental biochemical process conserved through evolution.

  19. Combined glutathione S transferase M1/T1 null genotypes is associated with type 2 diabetes mellitus

    Science.gov (United States)

    POROJAN, MIHAI D.; BALA, CORNELIA; ILIES, ROXANA; CATANA, ANDREEA; POPP, RADU A.; DUMITRASCU, DAN L.

    2015-01-01

    Background Due to new genetic insights, a considerably large number of genes and polymorphic gene variants are screened and linked with the complex pathogenesis of type 2 diabetes (DM). Our study aimed to investigate the association between the two isoforms of the glutathione S-transferase genes (Glutathione S transferase isoemzyme type M1- GSTM1 and Glutathione S transferase isoemzyme type T1-GSTT1) and the prevalence of DM in the Northern Romanian population. Methods We conducted a cross-sectional, randomized, case-control study evaluating the frequency of GSTM1 and GSTT1 null alleles in patients diagnosed with DM. A total of 106 patients diagnosed with DM and 124 healthy controls were included in the study. GSTM1 and GSTT1 null alleles genotyping was carried out using Multiplex PCR amplification of relevant gene fragments, followed by gel electrophoresis analysis of the resulting amplicons. Results Molecular analysis did not reveal an increased frequency of the null GSTM1 and GSTT1 alleles (mutant genotypes) respectively in the DM group compared to controls (p=0.171, OR=1.444 CI=0.852–2.447; p=0.647, OR=0.854, CI=0.436–1.673). Nevertheless, the combined GSTM1/GSTT1 null genotypes were statistically significantly higher in DM patients compared to control subjects (p=0.0021, OR=0.313, CI=0.149–0.655) Conclusions The main finding of our study is that the combined, double GSTM1/GSTT1 null genotypes are to be considered among the polymorphic genetic risk factors for type 2 DM. PMID:26528065

  20. The Fusarium oxysporum gnt2, Encoding a Putative N-Acetylglucosamine Transferase, Is Involved in Cell Wall Architecture and Virulence

    Science.gov (United States)

    López-Fernández, Loida; Ruiz-Roldán, Carmen; Pareja-Jaime, Yolanda; Prieto, Alicia; Khraiwesh, Husam; Roncero, M. Isabel G.

    2013-01-01

    With the aim to decipher the molecular dialogue and cross talk between Fusarium oxysporum f.sp. lycopersci and its host during infection and to understand the molecular bases that govern fungal pathogenicity, we analysed genes presumably encoding N-acetylglucosaminyl transferases, involved in glycosylation of glycoproteins, glycolipids, proteoglycans or small molecule acceptors in other microorganisms. In silico analysis revealed the existence of seven putative N-glycosyl transferase encoding genes (named gnt) in F. oxysporum f.sp. lycopersici genome. gnt2 deletion mutants showed a dramatic reduction in virulence on both plant and animal hosts. Δgnt2 mutants had αalterations in cell wall properties related to terminal αor β-linked N-acetyl glucosamine. Mutant conidia and germlings also showed differences in structure and physicochemical surface properties. Conidial and hyphal aggregation differed between the mutant and wild type strains, in a pH independent manner. Transmission electron micrographs of germlings showed strong cell-to-cell adherence and the presence of an extracellular chemical matrix. Δgnt2 cell walls presented a significant reduction in N-linked oligosaccharides, suggesting the involvement of Gnt2 in N-glycosylation of cell wall proteins. Gnt2 was localized in Golgi-like sub-cellular compartments as determined by fluorescence microscopy of GFP::Gnt2 fusion protein after treatment with the antibiotic brefeldin A or by staining with fluorescent sphingolipid BODIPY-TR ceramide. Furthermore, density gradient ultracentrifugation allowed co-localization of GFP::Gnt2 fusion protein and Vps10p in subcellular fractions enriched in Golgi specific enzymatic activities. Our results suggest that N-acetylglucosaminyl transferases are key components for cell wall structure and influence interactions of F. oxysporum with both plant and animal hosts during pathogenicity. PMID:24416097

  1. The Fusarium oxysporum gnt2, encoding a putative N-acetylglucosamine transferase, is involved in cell wall architecture and virulence.

    Directory of Open Access Journals (Sweden)

    Loida López-Fernández

    Full Text Available With the aim to decipher the molecular dialogue and cross talk between Fusarium oxysporum f.sp. lycopersci and its host during infection and to understand the molecular bases that govern fungal pathogenicity, we analysed genes presumably encoding N-acetylglucosaminyl transferases, involved in glycosylation of glycoproteins, glycolipids, proteoglycans or small molecule acceptors in other microorganisms. In silico analysis revealed the existence of seven putative N-glycosyl transferase encoding genes (named gnt in F. oxysporum f.sp. lycopersici genome. gnt2 deletion mutants showed a dramatic reduction in virulence on both plant and animal hosts. Δgnt2 mutants had αalterations in cell wall properties related to terminal αor β-linked N-acetyl glucosamine. Mutant conidia and germlings also showed differences in structure and physicochemical surface properties. Conidial and hyphal aggregation differed between the mutant and wild type strains, in a pH independent manner. Transmission electron micrographs of germlings showed strong cell-to-cell adherence and the presence of an extracellular chemical matrix. Δgnt2 cell walls presented a significant reduction in N-linked oligosaccharides, suggesting the involvement of Gnt2 in N-glycosylation of cell wall proteins. Gnt2 was localized in Golgi-like sub-cellular compartments as determined by fluorescence microscopy of GFP::Gnt2 fusion protein after treatment with the antibiotic brefeldin A or by staining with fluorescent sphingolipid BODIPY-TR ceramide. Furthermore, density gradient ultracentrifugation allowed co-localization of GFP::Gnt2 fusion protein and Vps10p in subcellular fractions enriched in Golgi specific enzymatic activities. Our results suggest that N-acetylglucosaminyl transferases are key components for cell wall structure and influence interactions of F. oxysporum with both plant and animal hosts during pathogenicity.

  2. Alterations at the peptidyl transferase centre of the ribosome induced by the synergistic action of the streptogramins dalfopristin and quinupristin

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    Fucini Paola

    2004-04-01

    Full Text Available Abstract Background The bacterial ribosome is a primary target of several classes of antibiotics. Investigation of the structure of the ribosomal subunits in complex with different antibiotics can reveal the mode of inhibition of ribosomal protein synthesis. Analysis of the interactions between antibiotics and the ribosome permits investigation of the specific effect of modifications leading to antimicrobial resistances. Streptogramins are unique among the ribosome-targeting antibiotics because they consist of two components, streptogramins A and B, which act synergistically. Each compound alone exhibits a weak bacteriostatic activity, whereas the combination can act bactericidal. The streptogramins A display a prolonged activity that even persists after removal of the drug. However, the mode of activity of the streptogramins has not yet been fully elucidated, despite a plethora of biochemical and structural data. Results The investigation of the crystal structure of the 50S ribosomal subunit from Deinococcus radiodurans in complex with the clinically relevant streptogramins quinupristin and dalfopristin reveals their unique inhibitory mechanism. Quinupristin, a streptogramin B compound, binds in the ribosomal exit tunnel in a similar manner and position as the macrolides, suggesting a similar inhibitory mechanism, namely blockage of the ribosomal tunnel. Dalfopristin, the corresponding streptogramin A compound, binds close to quinupristin directly within the peptidyl transferase centre affecting both A- and P-site occupation by tRNA molecules. Conclusions The crystal structure indicates that the synergistic effect derives from direct interaction between both compounds and shared contacts with a single nucleotide, A2062. Upon binding of the streptogramins, the peptidyl transferase centre undergoes a significant conformational transition, which leads to a stable, non-productive orientation of the universally conserved U2585. Mutations of this r

  3. Expression of an enzymatically active Yb3 glutathione S-transferase in Escherichia coli and identification of its natural form in rat brain.

    Science.gov (United States)

    Abramovitz, M; Ishigaki, S; Felix, A M; Listowsky, I

    1988-11-25

    Glutathione S-transferases containing Yb3 subunits are relatively uncommon forms that are expressed in a tissue-specific manner and have not been identified unequivocally or characterized. A cDNA clone containing the entire coding sequence of Yb3 glutathione S-transferase mRNA was incorporated into a pIN-III expression vector used to transform Escherichia coli. A fusion Yb3-protein containing 14 additional amino acid residues at its N terminus was purified to homogeneity. Recombinant Yb3 was enzymatically active with both 1-chloro-2,4-dinitrobenzene and 1,2-dichloro-4-nitrobenzene as substrates but lacked glutathione peroxidase activity. Substrate specificity patterns of recombinant Yb3 were more limited than those of glutathione S-transferase isoenzymes containing Yb1- or Yb2-type subunits. Peptides corresponding to unique amino acid sequences of Yb3 as well as a peptide from a region of homology with Yb1 and Yb2 subunits were synthesized. These synthetic peptides were used to raise antibodies specific to Yb3 and others that cross-reacted with all Yb forms. Immunoblotting was utilized to identify the natural counterpart of recombinant Yb3 among rat glutathione transferases. Brain and testis glutathione S-transferases were rich in Yb3 subunits, but very little was found in liver or kidney. Physical properties, substrate specificities, and binding patterns of the recombinant protein paralleled properties of the natural isoenzyme isolated from brain.

  4. Ameliorating Effects of Biochar Derived from Poultry Manure and White Clover Residues on Soil Nutrient Status and Plant growth Promotion--Greenhouse Experiments.

    Science.gov (United States)

    Abbasi, M Kaleem; Anwar, Ahsan Ali

    2015-01-01

    Biochar application to agricultural soils is rapidly emerging as a new management strategy for its potential role in carbon sequestration, soil quality improvements, and plant growth promotion. The aim of our study was to investigate the effects of biochars derived from white clover residues and poultry manure on soil quality characteristics, growth and N accumulation in maize (Zea mays L.) and wheat (Triticum aestivum L.) grown in a loam soil under greenhouse conditions. Treatments comprised of: untreated control; mineral N fertilizer (urea N, UN) at the rate of 200, and 100 mg N kg(-1), white clover residues biochar (WCRB), poultry manure biochar (PMB) at 30 Mg ha(-1), and the possible combinations of WCRB+PMB (50:50), UN+WCRB (50:50), UN+PMB (50:50), and UN+WCRB+PMB (50:25:25). The treatments were arranged in a completely randomized design with three replications. Results indicated a significant increase in the growth and biomass production of maize and wheat supplemented with biochars alone or mixed with N fertilizer. Biochars treatments showed varying impact on plant growth depended upon the type of the biochar, and in general plant growth under PMB was significantly higher than that recorded under WCRB. The growth characteristics in the combined treatments (half biochar+half N) were either higher or equivalent to that recorded under full fertilizer N treatment (N200). The biochar treatments WCRB, PMB, and WCRB+PMB (50:50) increased maize shoot N by 18, 26 and 21%, respectively compared to the control while wheat shoot N did not show positive response. The N-uptake by maize treated with WCRB, PMB, and WCRB+PMB (50:50) was 54, 116, and 90 mg g(-1) compared to the 33 mg g(-1) in the control while the N-uptake by wheat was 41, 60, and 53 mg g(-1) compared to 24 mg g(-1) in the control. The mixed treatments (half biochar+half N) increased N-uptake by 2.3 folds in maize and 1.7 to 2.5 folds in wheat compared to the N100 showing increasing effect of biochar on N

  5. Produtividade de sistemas forrageiros consorciados com amendoim forrageiro ou trevo vermelho Productivity of pastures-based systems mixed to forage peanut or red clover

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    Ricardo Lima de Azevedo Junior

    2012-11-01

    Full Text Available O objetivo desta pesquisa foi avaliar três sistemas forrageiros (SF com capim elefante (CE Pennisetum purpureum Schum., cv. 'MerckeronPinda' + espécies de crescimento espontâneo (ECE + azevém anual (AZ Lolium multiflorum Lam., cv. 'Comum', como SF1; CE + ECE + AZ + amendoim forrageiro (AF Arachis pintoi Krap. e Greg., cv. 'Amarillo', como SF2; e CE + ECE + AZ + trevo vermelho (TV Trifolium pratense L., cv. 'Estanzoela 116', como SF3. O CE foi estabelecido em linhas afastadas a cada 4m. O azevém anual foi estabelecido entre as linhas do CE durante o período hibernal; o TV foi semeado e o AF foi preservado nos respectivos tratamentos. Para avaliação, foram usadas vacas da raça Holandesa que receberam 5,5kg dia-1 como complemento alimentar. Foram avaliadas a taxa de acúmulo diário de matéria seca (TA, a massa de forragem desaparecida (MFD, a matéria seca desaparecida com base em 100kg de peso vivo (MSD e a produção de forragem (PF, as composições botânica e estrutural do CE. O delineamento experimental foi o inteiramente casualizado, com três tratamentos (SF e duas repetições (piquetes em parcelas subdivididas no tempo (pastejo. Durante o período experimental (341 dias, foram efetuados nove ciclos de pastejo. Os valores médios de TA, MFD, MSD e PF foram de 53,16kg ha-1; 36,13%; 2,77kg de matéria seca por 100kg de peso vivo e 17,80t ha-1. Para a variável ECE, houve aumento significativo no SF1. Considerando a carga animal, o SF3 apresentou melhor desempenho.The objective of this research was to evaluate of tree pasture-based systems (PS with elephantgrass (EG Pennisetum purpureum Schum., cv. 'Merckeron Pinda' + spontaneous growing species (SGS, annual ryegrass (RG Lolium multiflorum Lam., cv. 'Comum', for PS1; EG + SGS + forage peanut (FP Arachis pintoi Krap. e Greg., cv. 'Amarillo', for PS2; and EG + SGS + RG + red clover (RC Trifolium pratense L., cv. 'Estanzoela 116', for PS3. EG was planted in lines with a distance of 4m

  6. Does occupational exposure to solvents and pesticides in association with glutathione S-transferase A1, M1, P1, and T1 polymorphisms increase the risk of bladder cancer? The Belgrade case-control study.

    Directory of Open Access Journals (Sweden)

    Marija G Matic

    Full Text Available OBJECTIVE: We investigated the role of the glutathione S-transferase A1, M1, P1 and T1 gene polymorphisms and potential effect modification by occupational exposure to different chemicals in Serbian bladder cancer male patients. PATIENTS AND METHODS: A hospital-based case-control study of bladder cancer in men comprised 143 histologically confirmed cases and 114 age-matched male controls. Deletion polymorphism of glutathione S-transferase M1 and T1 was identified by polymerase chain reaction method. Single nucleotide polymorphism of glutathione S-transferase A1 and P1 was identified by restriction fragment length polymorphism method. As a measure of effect size, odds ratio (OR with corresponding 95% confidence interval (95%CI was calculated. RESULTS: The glutathione S-transferase A1, T1 and P1 genotypes did not contribute independently toward the risk of bladder cancer, while the glutathione S-transferase M1-null genotype was overrepresented among cases (OR = 2.1, 95% CI = 1.1-4.2, p = 0.032. The most pronounced effect regarding occupational exposure to solvents and glutathione S-transferase genotype on bladder cancer risk was observed for the low activity glutathione S-transferase A1 genotype (OR = 9.2, 95% CI = 2.4-34.7, p = 0.001. The glutathione S-transferase M1-null genotype also enhanced the risk of bladder cancer among subjects exposed to solvents (OR = 6,5, 95% CI = 2.1-19.7, p = 0.001. The risk of bladder cancer development was 5.3-fold elevated among glutathione S-transferase T1-active patients exposed to solvents in comparison with glutathione S-transferase T1-active unexposed patients (95% CI = 1.9-15.1, p = 0.002. Moreover, men with glutathione S-transferase T1-active genotype exposed to pesticides exhibited 4.5 times higher risk in comparison with unexposed glutathione S-transferase T1-active subjects (95% CI = 0.9-22.5, p = 0.067. CONCLUSION: Null or low-activity genotypes of the

  7. Cap-independent translation mechanism of red clover necrotic mosaic virus RNA2 differs from that of RNA1 and is linked to RNA replication.

    Science.gov (United States)

    Mizumoto, Hiroyuki; Iwakawa, Hiro-Oki; Kaido, Masanori; Mise, Kazuyuki; Okuno, Tetsuro

    2006-04-01

    The genome of Red clover necrotic mosaic virus (RCNMV) in the genus Dianthovirus is divided into two RNA molecules of RNA1 and RNA2, which have no cap structure at the 5' end and no poly(A) tail at the 3' end. The 3' untranslated region (3' UTR) of RCNMV RNA1 contains an essential RNA element (3'TE-DR1), which is required for cap-independent translation. In this study, we investigated a cap-independent translational mechanism of RNA2 using a firefly luciferase (Luc) gene expression assay system in cowpea protoplasts and a cell-free lysate (BYL) prepared from evacuolated tobacco BY2 protoplasts. We were unable to detect cis-acting RNA sequences in RNA2 that can replace the function of a cap structure, such as the 3'TE-DR1 of RNA1. However, the uncapped reporter RNA2, RNA2-Luc, in which the Luc open reading frame (ORF) was inserted between the 5' UTR and the movement protein ORF, was effectively translated in the presence of p27 and p88 in protoplasts in which RNA2-Luc was replicated. Time course experiments in protoplasts showed that the translational activity of RNA2-Luc did not reflect the amount of RNA2. Mutations in cis-acting RNA replication elements of RNA2 abolished the cap-independent translational activity of RNA2-Luc, suggesting that the translational activity of RNA2-Luc is coupled to RNA replication. Our results show that the translational mechanism differs between two segmented genomic RNAs of RCNMV. We present a model in which only RNA2 that is generated de novo through the viral RNA replication machinery functions as mRNA for translation.

  8. Polyamine regulates tolerance to water stress in leaves of white clover associated with antioxidant defense and dehydrin genes via involvement in calcium messenger system and hydrogen peroxide signaling

    Directory of Open Access Journals (Sweden)

    Zhou eLi

    2015-10-01

    Full Text Available Endogenous polyamine (PA may play a critical role in tolerance to water stress in plants acting as a signaling molecule activator. Water stress caused increases in endogenous PA content in leaves, including putrescine (Put, spermidine (Spd, and spermine (Spm. Exogenous application of Spd could induce the instantaneous H2O2 burst and accumulation of cytosolic free Ca2+, and activate NADPH oxidase and CDPK gene expression in cells. To a great extent, PA biosynthetic inhibitor reduced the water stress-induced H2O2 accumulation, free cytosolic Ca2+ release, antioxidant enzyme activities and genes expression leading to aggravate water stress-induced oxidative damage, while these suppressing effects were alleviated by the addition of exogenous Spd, indicating PA was involved in water stress-induced H2O2 and cytosolic free Ca2+ production as well as stress tolerance. Dehydrin genes (Y2SK, Y2K, and SK2 were showed to be highly responsive to exogenous Spd. PA-induced antioxidant defense and dehydrin genes expression could be blocked by the scavenger of H2O2 and the inhibitors of H2O2 generation or Ca2+ channels blockers, a calmodulin antagonist, as well as the inhibitor of CDPK. These findings suggested that PA regulated tolerance to water stress in white clover associated with antioxidant defenses and dehydrins via involvement in the calcium messenger system and H2O2 signaling pathways. PA-induced H2O2 production required Ca2+ release, while PA-induced Ca2+ release was also essential for H2O2 production, suggesting an interaction between PA-induced H2O2 and Ca2+ signaling.

  9. Changes in soil solution Zn and pH and uptake of Zn by arbuscular mycorrhizal red clover in Zn-contaminated soil.

    Science.gov (United States)

    Li, X; Christie, P

    2001-01-01

    Red clover plants inoculated with Glomus mosseae were grown in a sterile pasture soil containing 50 mg Zn kg(-1) in 'Plexiglas' (acrylic) containers with nylon net partitions (30 microm mesh) designed to separate the soil into a central root zone and two outer zones for hyphal growth with no root penetration. Two porous plastic soil moisture samplers were installed in each pot, one in the root compartment and the other in one of the hyphal compartments. The soil in the outer compartments was amended with one of the four application rates of Zn (as ZnSO4) ranging from 0 to 1000 mg kg(-1). Non-mycorrhizal controls were included, and there were five replicates of each treatment in a randomised block in a glasshouse. Uninoculated plants received supplementary P to avoid yield limitation due to low soil P status. Plants grew in the central compartment for nine weeks. Soil moisture samples were collected 4, 24 and 62 days after sowing to monitor changes in the Zn concentration and pH of the soil solution. At harvest, the mean mycorrhizal infection rate of inoculated plants ranged from 29% to 34% of total root length and was little affected by Zn application. Root and shoot yields were not affected by mycorrhizal infection. Plant Zn concentration and uptake were lower in mycorrhizal plants than non-mycorrhizal controls, and this effect was more pronounced with increasing Zn application rate to the soil. Soil solution Zn concentrations were lower and pH values were higher in mycorrhizal treatments than non-mycorrhizal controls and the mycorrhiza effect was more pronounced at higher Zn application rates. The protective effect of mycorrhiza against plant Zn uptake may have been associated with changes in Zn solubility mediated by changes in the soil solution pH, or by immobilisation of Zn in the extraradical mycelium.

  10. Crystallization and preliminary X-ray diffraction analysis of a glutathione S-transferase from Xylella fastidiosa

    International Nuclear Information System (INIS)

    Garcia, Wanius; Travensolo, Regiane F.; Rodrigues, Nathalia C.; Muniz, João R. C.; Caruso, Célia S.; Lemos, Eliana G. M.; Araujo, Ana Paula U.; Carrilho, Emanuel

    2008-01-01

    Glutathione S-transferase from X. fastidiosa (xfGST) has been overexpressed in E. coli, purified and crystallized. Diffraction data were collected to 2.23 Å. Glutathione S-transferases (GSTs) form a group of multifunctional isoenzymes that catalyze the glutathione-dependent conjugation and reduction reactions involved in the cellular detoxification of xenobiotic and endobiotic compounds. GST from Xylella fastidiosa (xfGST) was overexpressed in Escherichia coli and purified by conventional affinity chromatography. In this study, the crystallization and preliminary X-ray analysis of xfGST is described. The purified protein was crystallized by the vapour-diffusion method, producing crystals that belonged to the triclinic space group P1. The unit-cell parameters were a = 47.73, b = 87.73, c = 90.74 Å, α = 63.45, β = 80.66, γ = 94.55°. xfGST crystals diffracted to 2.23 Å resolution on a rotating-anode X-ray source

  11. Glutathione transferase (GST) as a candidate molecular-based biomarker for soil toxin exposure in the earthworm Lumbricus rubellus

    International Nuclear Information System (INIS)

    LaCourse, E. James; Hernandez-Viadel, Mariluz; Jefferies, James R.; Svendsen, Claus; Spurgeon, David J.; Barrett, John; John Morgan, A.; Kille, Peter; Brophy, Peter M.

    2009-01-01

    The earthworm Lumbricus rubellus (Hoffmeister, 1843) is a terrestrial pollution sentinel. Enzyme activity and transcription of phase II detoxification superfamily glutathione transferases (GST) is known to respond in earthworms after soil toxin exposure, suggesting GST as a candidate molecular-based pollution biomarker. This study combined sub-proteomics, bioinformatics and biochemical assay to characterise the L. rubellus GST complement as pre-requisite to initialise assessment of the applicability of GST as a biomarker. L. rubellus possesses a range of GSTs related to known classes, with evidence of tissue-specific synthesis. Two affinity-purified GSTs dominating GST protein synthesis (Sigma and Pi class) were cloned, expressed and characterised for enzyme activity with various substrates. Electrospray ionisation mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) following SDS-PAGE were superior in retaining subunit stability relative to two-dimensional gel electrophoresis (2-DE). This study provides greater understanding of Phase II detoxification GST superfamily status of an important environmental pollution sentinel organism. - This study currently provides the most comprehensive view of the Phase II detoxification enzyme superfamily of glutathione transferases within the important environmental pollution sentinel earthworm Lumbricus rubellus.

  12. Glutathione transferase (GST) as a candidate molecular-based biomarker for soil toxin exposure in the earthworm Lumbricus rubellus

    Energy Technology Data Exchange (ETDEWEB)

    LaCourse, E. James, E-mail: james.la-course@liverpool.ac.u [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom); Hernandez-Viadel, Mariluz; Jefferies, James R. [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom); Svendsen, Claus; Spurgeon, David J. [Centre for Ecology and Hydrology, Huntingdon PE28 2LS (United Kingdom); Barrett, John [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom); John Morgan, A.; Kille, Peter [Biosciences, University of Cardiff, Cardiff CF10 3TL (United Kingdom); Brophy, Peter M. [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom)

    2009-08-15

    The earthworm Lumbricus rubellus (Hoffmeister, 1843) is a terrestrial pollution sentinel. Enzyme activity and transcription of phase II detoxification superfamily glutathione transferases (GST) is known to respond in earthworms after soil toxin exposure, suggesting GST as a candidate molecular-based pollution biomarker. This study combined sub-proteomics, bioinformatics and biochemical assay to characterise the L. rubellus GST complement as pre-requisite to initialise assessment of the applicability of GST as a biomarker. L. rubellus possesses a range of GSTs related to known classes, with evidence of tissue-specific synthesis. Two affinity-purified GSTs dominating GST protein synthesis (Sigma and Pi class) were cloned, expressed and characterised for enzyme activity with various substrates. Electrospray ionisation mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) following SDS-PAGE were superior in retaining subunit stability relative to two-dimensional gel electrophoresis (2-DE). This study provides greater understanding of Phase II detoxification GST superfamily status of an important environmental pollution sentinel organism. - This study currently provides the most comprehensive view of the Phase II detoxification enzyme superfamily of glutathione transferases within the important environmental pollution sentinel earthworm Lumbricus rubellus.

  13. The role of human demographic history in determining the distribution and frequency of transferase-deficient galactosaemia mutations.

    LENUS (Irish Health Repository)

    Flanagan, J M

    2010-02-01

    Classical or transferase-deficient galactosaemia is an inherited metabolic disorder caused by mutation in the human Galactose-1-phosphate uridyl transferase (GALT) gene. Of some 170 causative mutations reported, fewer than 10% are observed in more than one geographic region or ethnic group. To better understand the population history of the common GALT mutations, we have established a haplotyping system for the GALT locus incorporating eight single nucleotide polymorphisms and three short tandem repeat markers. We analysed haplotypes associated with the three most frequent GALT gene mutations, Q188R, K285N and Duarte-2 (D2), and estimated their age. Haplotype diversity, in conjunction with measures of genetic diversity and of linkage disequilibrium, indicated that Q188R and K285N are European mutations. The Q188R mutation arose in central Europe within the last 20 000 years, with its observed east-west cline of increasing relative allele frequency possibly being due to population expansion during the re-colonization of Europe by Homo sapiens in the Mesolithic age. K285N was found to be a younger mutation that originated in Eastern Europe and is probably more geographically restricted as it arose after all major European population expansions. The D2 variant was found to be an ancient mutation that originated before the expansion of Homo sapiens out of Africa.

  14. Acute cadmium intoxication induces alpha-class glutathione S-transferase protein synthesis and enzyme activity in rat liver

    International Nuclear Information System (INIS)

    Casalino, Elisabetta; Sblano, Cesare; Calzaretti, Giovanna; Landriscina, Clemente

    2006-01-01

    Acute cadmium intoxication affects glutathione S-transferase (GST) in rat liver. It has been found that 24 h after i.p. cadmium administration to rats, at a dose of 2.5 mg CdCl 2 kg -1 body weight, the activity of this enzyme in liver cytosol increased by 40%. A less stimulatory effect persisted till 48 h and thereafter the enzyme activity normalized. Since, GST isoenzymes belong to different classes in mammalian tissues, we used quantitative immunoassays to verify which family of GST isoenzymes is influenced by this intoxication. Only alpha-class glutathione S-transferase (α-GST) proteins were detected in rat liver cytosol and their level increased by about 25%, 24 h after cadmium treatment. No pi-GST isoforms were found in liver cytosol from either normal or cadmium-treated rats. Co-administration of actinomycin D with cadmium normalized both the protein level and the activity of α-GST, suggesting that some effect occurs on enzyme transcription of these isoenzymes by this metal. On the other hand, it seems unlikely that the stimulatory effect is due to the high level of peroxides caused by lipid peroxidation, since Vitamin E administration strongly reduced the TBARS level, but did not cause any GST activity decrease

  15. Crystallization and preliminary X-ray diffraction analysis of a glutathione S-transferase from Xylella fastidiosa

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, Wanius, E-mail: wanius@if.sc.usp.br [Laboratório de Biofísica Molecular ‘Sérgio Mascarenhas’, Instituto de Física de São Carlos, Universidade de São Paulo (USP), São Carlos (Brazil); Travensolo, Regiane F. [Grupo de Bioanalítica, Microfabricação e Separações, Instituto de Química de São Carlos, Universidade de São Paulo (USP), São Carlos (Brazil); Rodrigues, Nathalia C.; Muniz, João R. C. [Laboratório de Biofísica Molecular ‘Sérgio Mascarenhas’, Instituto de Física de São Carlos, Universidade de São Paulo (USP), São Carlos (Brazil); Caruso, Célia S. [Grupo de Bioanalítica, Microfabricação e Separações, Instituto de Química de São Carlos, Universidade de São Paulo (USP), São Carlos (Brazil); Lemos, Eliana G. M. [Laboratório de Bioquímica de Microrganismos e de Plantas, Departamento de Tecnologia, UNESP, Jaboticabal (Brazil); Araujo, Ana Paula U. [Laboratório de Biofísica Molecular ‘Sérgio Mascarenhas’, Instituto de Física de São Carlos, Universidade de São Paulo (USP), São Carlos (Brazil); Carrilho, Emanuel, E-mail: wanius@if.sc.usp.br [Grupo de Bioanalítica, Microfabricação e Separações, Instituto de Química de São Carlos, Universidade de São Paulo (USP), São Carlos (Brazil); Laboratório de Biofísica Molecular ‘Sérgio Mascarenhas’, Instituto de Física de São Carlos, Universidade de São Paulo (USP), São Carlos (Brazil)

    2008-02-01

    Glutathione S-transferase from X. fastidiosa (xfGST) has been overexpressed in E. coli, purified and crystallized. Diffraction data were collected to 2.23 Å. Glutathione S-transferases (GSTs) form a group of multifunctional isoenzymes that catalyze the glutathione-dependent conjugation and reduction reactions involved in the cellular detoxification of xenobiotic and endobiotic compounds. GST from Xylella fastidiosa (xfGST) was overexpressed in Escherichia coli and purified by conventional affinity chromatography. In this study, the crystallization and preliminary X-ray analysis of xfGST is described. The purified protein was crystallized by the vapour-diffusion method, producing crystals that belonged to the triclinic space group P1. The unit-cell parameters were a = 47.73, b = 87.73, c = 90.74 Å, α = 63.45, β = 80.66, γ = 94.55°. xfGST crystals diffracted to 2.23 Å resolution on a rotating-anode X-ray source.

  16. Activity-Based Probes for Isoenzyme- and Site-Specific Functional Characterization of Glutathione S -Transferases

    Energy Technology Data Exchange (ETDEWEB)

    Stoddard, Ethan G. [Chemical Biology and Exposure; Killinger, Bryan J. [Chemical Biology and Exposure; Nair, Reji N. [Chemical Biology and Exposure; Sadler, Natalie C. [Chemical Biology and Exposure; Volk, Regan F. [Chemical Biology and Exposure; Purvine, Samuel O. [Chemical Biology and Exposure; Shukla, Anil K. [Chemical Biology and Exposure; Smith, Jordan N. [Chemical Biology and Exposure; Wright, Aaron T. [Chemical Biology and Exposure

    2017-11-01

    Glutathione S-transferases (GSTs) comprise a highly diverse family of phase II drug metabolizing enzymes whose shared function is the conjugation of reduced glutathione to various endo- and xenobiotics. Although the conglomerate activity of these enzymes can be measured by colorimetric assays, measurement of the individual contribution from specific isoforms and their contribution to the detoxification of xenobiotics in complex biological samples has not been possible. For this reason, we have developed two activity-based probes that characterize active glutathione transferases in mammalian tissues. The GST active site is comprised of a glutathione binding “G site” and a distinct substrate binding “H site”. Therefore, we developed (1) a glutathione-based photoaffinity probe (GSH-ABP) to target the “G site”, and (2) a probe designed to mimic a substrate molecule and show “H site” activity (GST-ABP). The GSH-ABP features a photoreactive moiety for UV-induced covalent binding to GSTs and glutathione-binding enzymes. The GST-ABP is a derivative of a known mechanism-based GST inhibitor that binds within the active site and inhibits GST activity. Validation of probe targets and “G” and “H” site specificity was carried out using a series of competitors in liver homogenates. Herein, we present robust tools for the novel characterization of enzyme- and active site-specific GST activity in mammalian model systems.

  17. Cantharidin Impedes Activity of Glutathione S-Transferase in the Midgut of Helicoverpa armigera Hübner

    Directory of Open Access Journals (Sweden)

    Ya Lin Zhang

    2013-03-01

    Full Text Available Previous investigations have implicated glutathione S-transferases (GSTs as one of the major reasons for insecticide resistance. Therefore, effectiveness of new candidate compounds depends on their ability to inhibit GSTs to prevent metabolic detoxification by insects. Cantharidin, a terpenoid compound of insect origin, has been developed as a bio-pesticide in China, and proves highly toxic to a wide range of insects, especially lepidopteran. In the present study, we test cantharidin as a model compound for its toxicity, effects on the mRNA transcription of a model Helicoverpa armigera glutathione S-transferase gene (HaGST and also for its putative inhibitory effect on the catalytic activity of GSTs, both in vivo and in vitro in Helicoverpa armigera, employing molecular and biochemical methods. Bioassay results showed that cantharidin was highly toxic to H. armigera. Real-time qPCR showed down-regulation of the HaGST at the mRNA transcript ranging from 2.5 to 12.5 folds while biochemical assays showed in vivo inhibition of GSTs in midgut and in vitro inhibition of rHaGST. Binding of cantharidin to HaGST was rationalized by homology and molecular docking simulations using a model GST (1PN9 as a template structure. Molecular docking simulations also confirmed accurate docking of the cantharidin molecule to the active site of HaGST impeding its catalytic activity.

  18. Glutathione S transferase polymorphisms influence on iron overload in β-thalassemia patients

    Directory of Open Access Journals (Sweden)

    Serena Sclafani

    2013-11-01

    Full Text Available In patients with β-thalassemia iron overload that leads to damage to vital organs is observed. Glutathione S transferase (GST enzymes have an antioxidant role in detoxification processes of toxic substances. This role is determined genetically. In this study, we correlated GSTT1 and GSTM1 genotypes with iron overload measured with direct and indirect non-invasive methods; in particular, we used serum ferritin and signal intensity of the magnetic resonance image (MRI in 42 patients with β-thalassemia, which were regularly subjected to chelation and transfusion therapy. Multiplex polymerase chain reaction was used to determine the genotype. The loss of both alleles leads to a decreased value of liver and heart MRI-signal intensity with a consequent iron accumulation in these organs; the loss of only one allele doesn’t lead to relevant overload. Serum ferritin doesn’t appear to be correlated to iron overload instead. 对于β-地中海贫血患者,由于铁过量而造成重要器官受损的情况也在观察之中。谷胱甘肽S转移酶(GST 酶类在对有毒物质进行解毒的过程中有着抗氧化剂的作用。该作用是由基因决定的。 在这份研究中,我们运用了直接和间接非侵入性的方法对基因型铁过量GSTT1 和GSTM1进行了相关性测量;特别地,我们对42位定期接受螯合和输血治疗的β-地中海贫血患者进行了血清铁蛋白和磁共振强度图像(MRI 的测试。 多重聚合酶链反应的测试也被运用来确定该基因型。 该两种等位基因的缺失,导致了肝功能减损及心脏磁共振强度的下降,并造成了在这些器官中铁含量的积累;其中一种等位基因的缺失并不会导致过度的铁含量。血清蛋白和铁过量之间,看起来并不存在相关性。

  19. A comparative study of transfer factors of water, iodine and strontium on rye-grass and clover. Development of a model of evaluation of the limits of foliar contamination by wet deposit

    International Nuclear Information System (INIS)

    Angeletti, Livio; Levi, Emilio.

    1977-07-01

    Transfer factors of water, iodine ( 131 I) and strontium ( 85 Sr) on above-ground parts of rye-grass and clover were determined as a function of aspersion intensities. An analysis of the results showed the effect of aspersion intensities, nature of the chemical element and plant species on the values of transfer factors of iodine and strontium. It also made it possible to propose a simple method of evaluation of contamination limits of the aerial parts of plants by wet deposit, based on transfer values of water on plants only [fr

  20. The lectin domains of polypeptide GalNAc-transferases exhibit carbohydrate-binding specificity for GalNAc: lectin binding to GalNAc-glycopeptide substrates is required for high density GalNAc-O-glycosylation

    DEFF Research Database (Denmark)

    Wandall, Hans H; Irazoqui, Fernando; Tarp, Mads Agervig

    2007-01-01

    Initiation of mucin-type O-glycosylation is controlled by a large family of UDP GalNAc:polypeptide N-acetylgalactosaminyltransferases (GalNAc-transferases). Most GalNAc-transferases contain a ricin-like lectin domain in the C-terminal end, which may confer GalNAc-glycopeptide substrate specificit...

  1. Intracellular APP Domain Regulates Serine-Palmitoyl-CoA Transferase Expression and Is Affected in Alzheimer's Disease

    Science.gov (United States)

    Grimm, Marcus O. W.; Grösgen, Sven; Rothhaar, Tatjana L.; Burg, Verena K.; Hundsdörfer, Benjamin; Haupenthal, Viola J.; Friess, Petra; Müller, Ulrike; Fassbender, Klaus; Riemenschneider, Matthias; Grimm, Heike S.; Hartmann, Tobias

    2011-01-01

    Lipids play an important role as risk or protective factors in Alzheimer's disease (AD), a disease biochemically characterized by the accumulation of amyloid beta peptides (Aβ), released by proteolytic processing of the amyloid precursor protein (APP). Changes in sphingolipid metabolism have been associated to the development of AD. The key enzyme in sphingolipid de novo synthesis is serine-palmitoyl-CoA transferase (SPT). In the present study we identified a new physiological function of APP in sphingolipid synthesis. The APP intracellular domain (AICD) was found to decrease the expression of the SPT subunit SPTLC2, the catalytic subunit of the SPT heterodimer, resulting in that decreased SPT activity. AICD function was dependent on Fe65 and SPTLC2 levels are increased in APP knock-in mice missing a functional AICD domain. SPTLC2 levels are also increased in familial and sporadic AD postmortem brains, suggesting that SPT is involved in AD pathology. PMID:21660213

  2. Yeast one-hybrid system used to identify the binding proteins for rat glutathione S-transferase P enhancer I.

    Science.gov (United States)

    Liao, Ming-Xiang; Liu, Dong-Yuan; Zuo, Jin; Fang, Fu-De

    2002-03-01

    To detect the trans-factors specifically binding to the strong enhancer element (GPEI) in the upstream of rat glutathione S-transferase P (GST-P) gene. Yeast one-hybrid system was used to screen rat lung MATCHMAKER cDNA library to identify potential trans-factors that can interact with core sequence of GPEI(cGPEI). Electrophoresis mobility shift assay (EMSA) was used to analyze the binding of transfactors to cGPEI. cDNA fragments coding for the C-terminal part of the transcription factor c-Jun and rat adenine nucleotide translocator (ANT) were isolated. The binding of c-Jun and ANT to GPEI core sequence were confirmed. Rat c-jun transcriptional factor and ANT may interact with cGPEI. They could play an important role in the induced expression of GST-P gene.

  3. Glutathione S-transferase of brown planthoppers (Nilaparvata lugens) is essential for their adaptation to gramine-containing host plants.

    Science.gov (United States)

    Sun, Xiao-Qin; Zhang, Mao-Xin; Yu, Jing-Ya; Jin, Yu; Ling, Bing; Du, Jin-Ping; Li, Gui-Hua; Qin, Qing-Ming; Cai, Qing-Nian

    2013-01-01

    Plants have evolved complex processes to ward off attacks by insects. In parallel, insects have evolved mechanisms to thwart these plant defenses. To gain insight into mechanisms that mediate this arms race between plants and herbivorous insects, we investigated the interactions between gramine, a toxin synthesized by plants of the family Gramineae, and glutathione S transferase (GST), an enzyme found in insects that is known to detoxify xenobiotics. Here, we demonstrate that rice (Oryza sativa), a hydrophytic plant, also produces gramine and that rice resistance to brown planthoppers (Nilaparvata lugens, BPHs) is highly associated with in planta gramine content. We also show that gramine is a toxicant that causes BPH mortality in vivo and that knockdown of BPH GST gene nlgst1-1 results in increased sensitivity to diets containing gramine. These results suggest that the knockdown of key detoxification genes in sap-sucking insects may provide an avenue for increasing their sensitivity to natural plant-associated defense mechanisms.

  4. Dynamic interplay between catalytic and lectin domains of GalNAc-transferases modulates protein O-glycosylation

    DEFF Research Database (Denmark)

    Lira-Navarrete, Erandi; de Las Rivas, Matilde; Compañón, Ismael

    2015-01-01

    the first crystal structures of complexes of GalNAc-T2 with glycopeptides that together with enhanced sampling molecular dynamics simulations demonstrate a cooperative mechanism by which the lectin domain enables free acceptor sites binding of glycopeptides into the catalytic domain. Atomic force microscopy......Protein O-glycosylation is controlled by polypeptide GalNAc-transferases (GalNAc-Ts) that uniquely feature both a catalytic and lectin domain. The underlying molecular basis of how the lectin domains of GalNAc-Ts contribute to glycopeptide specificity and catalysis remains unclear. Here we present...... and small-angle X-ray scattering experiments further reveal a dynamic conformational landscape of GalNAc-T2 and a prominent role of compact structures that are both required for efficient catalysis. Our model indicates that the activity profile of GalNAc-T2 is dictated by conformational heterogeneity...

  5. Protective effect of lemongrass oil against dexamethasone induced hyperlipidemia in rats: possible role of decreased lecithin cholesterol acetyl transferase activity.

    Science.gov (United States)

    Kumar, V R Santhosh; Inamdar, Md Naseeruddin; Nayeemunnisa; Viswanatha, G L

    2011-08-01

    To evaluate the anti-hyperlipidemic activity of lemongrass oil against in dexamethasone induced hyperlipidemia in rats. Administration of dexamethasone was given at 10 mg/kg, sc. to the adult rats for 8 d induces hyperlipidemia characterized by marked increase in serum cholesterol and triglyceride levels along with increase in atherogenic index. Lemongrass oil (100 and 200 mg/kg, po.) treatment has showed significant inhibition against dexamethasone hyperlipidemia by maintaining the serum levels of cholesterol, triglycerides and atherogenic index near to the normal levels and the antihyperlipidemic effect of the lemongross oil was comparable with atorvastatin 10 mg/kg, po. The possible mechanism may be associated with decrease in lecithin cholesterol acetyl transferase (LCAT) activity. These results suggested that Lemon gross oil possess significant anti-hyperlipidemic activity. Copyright © 2011 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  6. A SERS protocol as a potential tool to access 6-mercaptopurine release accelerated by glutathione-S-transferase.

    Science.gov (United States)

    Wang, Ying; Sun, Jie; Yang, Qingran; Lu, Wenbo; Li, Yan; Dong, Jian; Qian, Weiping

    2015-11-21

    The developed method for monitoring GST, an important drug metabolic enzyme, could greatly facilitate researches on relative biological fields. In this work, we have developed a SERS technique to monitor the absorbance behaviour of 6-mercaptopurine (6-MP) and its glutathione-S-transferase (GST)-accelerated glutathione (GSH)-triggered release behaviour on the surface of gold nanoflowers (GNFs), using the GNFs as excellent SERS substrates. The SERS signal was used as an indicator of absorbance or release of 6-MP on the gold surface. We found that GST can accelerate GSH-triggered release behaviour of 6-MP from the gold surface. We speculated that GST catalyzes nucleophilic GSH to competitively bind with the electrophilic substance 6-MP. Experimental results have proved that the presented SERS protocol can be utilized as an effective tool for accessing the release of anticancer drugs.

  7. The identification and characterisation of a functional interaction between arginyl-tRNA-protein transferase and topoisomerase II

    International Nuclear Information System (INIS)

    Barker, Catherine R.; Mouchel, Nathalie A.P.; Jenkins, John R.

    2006-01-01

    Topoisomerase II is required for the viability of all eukaryotic cells. It plays important roles in DNA replication, recombination, chromosome segregation, and the maintenance of the nuclear scaffold. Proteins that interact with and regulate this essential enzyme are of great interest. To investigate the role of proteins interacting with the N-terminal domain of the Saccharomyces cerevisiae topoisomerase II, we used a yeast two-hybrid protein interaction screen. We identified an interaction between arginyl-tRNA-protein transferase (Ate1) and the N-terminal domain of the S. cerevisiae topoisomerase II, including the potential site of interaction. Ate1 is a component of the N-end rule protein degradation pathway which targets proteins for degradation. We also propose a previously unidentified role for Ate1 in modulating the level of topoisomerase II through the cell cycle

  8. Identification and suppression of the p-coumaroyl CoA:hydroxycinnamyl alcohol transferase in Zea mays L.

    Science.gov (United States)

    Marita, Jane M; Hatfield, Ronald D; Rancour, David M; Frost, Kenneth E

    2014-06-01

    Grasses, such as Zea mays L. (maize), contain relatively high levels of p-coumarates (pCA) within their cell walls. Incorporation of pCA into cell walls is believed to be due to a hydroxycinnamyl transferase that couples pCA to monolignols. To understand the role of pCA in maize development, the p-coumaroyl CoA:hydroxycinnamyl alcohol transferase (pCAT) was isolated and purified from maize stems. Purified pCAT was subjected to partial trypsin digestion, and peptides were sequenced by tandem mass spectrometry. TBLASTN analysis of the acquired peptide sequences identified a single full-length maize cDNA clone encoding all the peptide sequences obtained from the purified enzyme. The cDNA clone was obtained and used to generate an RNAi construct for suppressing pCAT expression in maize. Here we describe the effects of suppression of pCAT in maize. Primary screening of transgenic maize seedling leaves using a new rapid analytical platform was used to identify plants with decreased amounts of pCA. Using this screening method, mature leaves from fully developed plants were analyzed, confirming reduced pCA levels throughout plant development. Complete analysis of isolated cell walls from mature transgenic stems and leaves revealed that lignin levels did not change, but pCA levels decreased and the lignin composition was altered. Transgenic plants with the lowest levels of pCA had decreased levels of syringyl units in the lignin. Thus, altering the levels of pCAT expression in maize leads to altered lignin composition, but does not appear to alter the total amount of lignin present in the cell walls. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  9. Synergistic and independent actions of multiple terminal nucleotidyl transferases in the 3' tailing of small RNAs in Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Xiaoyan Wang

    2015-04-01

    Full Text Available All types of small RNAs in plants, piwi-interacting RNAs (piRNAs in animals and a subset of siRNAs in Drosophila and C. elegans are subject to HEN1 mediated 3' terminal 2'-O-methylation. This modification plays a pivotal role in protecting small RNAs from 3' uridylation, trimming and degradation. In Arabidopsis, HESO1 is a major enzyme that uridylates small RNAs to trigger their degradation. However, U-tail is still present in null hen1 heso1 mutants, suggesting the existence of (an enzymatic activities redundant with HESO1. Here, we report that UTP: RNA uridylyltransferase (URT1 is a functional paralog of HESO1. URT1 interacts with AGO1 and plays a predominant role in miRNA uridylation when HESO1 is absent. Uridylation of miRNA is globally abolished in a hen1 heso1 urt1 triple mutant, accompanied by an extensive increase of 3'-to-5' trimming. In contrast, disruption of URT1 appears not to affect the heterochromatic siRNA uridylation. This indicates the involvement of additional nucleotidyl transferases in the siRNA pathway. Analysis of miRNA tailings in the hen1 heso1 urt1 triple mutant also reveals the existence of previously unknown enzymatic activities that can add non-uridine nucleotides. Importantly, we show HESO1 may also act redundantly with URT1 in miRNA uridylation when HEN1 is fully competent. Taken together, our data not only reveal a synergistic action of HESO1 and URT1 in the 3' uridylation of miRNAs, but also independent activities of multiple terminal nucleotidyl transferases in the 3' tailing of small RNAs and an antagonistic relationship between uridylation and trimming. Our results may provide further insight into the mechanisms of small RNA 3' end modification and stability control.

  10. Phylogenetic characterization of Clonorchis sinensis proteins homologous to the sigma-class glutathione transferase and their differential expression profiles.

    Science.gov (United States)

    Bae, Young-An; Kim, Jeong-Geun; Kong, Yoon

    2016-01-01

    Glutathione transferase (GST) is one of the major antioxidant proteins with diverse supplemental activities including peroxidase, isomerase, and thiol transferase. GSTs are classified into multiple classes on the basis of their primary structures and substrate/inhibitor specificity. However, the evolutionary routes and physiological environments specific to each of the closely related bioactive enzymes remain elusive. The sigma-like GSTs exhibit amino acid conservation patterns similar to the prostaglandin D synthases (PGDSs). In this study, we analyzed the phylogenetic position of the GSTs of the biocarcinogenic liver fluke, Clonorchis sinensis. We also observed induction profile of the GSTs in association with the parasite's maturation and in response to exogenous oxidative stresses, with special attention to sigma-class GSTs and PGDSs. The C. sinensis genome encoded 12 GST protein species, which were separately assigned to cytosolic (two omega-, one zeta-, two mu-, and five sigma-class), mitochondrial (one kappa-class), and microsomal (one membrane-associated proteins in eicosanoid and glutathione metabolism-like protein) GST families. Multiple sigma GST (or PGDS) orthologs were also detected in Opisthorchis viverrini. Other trematode species possessed only a single sigma-like GST gene. A phylogenetic analysis demonstrated that one of the sigma GST lineages duplicated in the common ancestor of trematodes were specifically expanded in the opisthorchiids, but deleted in other trematodes. The induction profiles of these sigma GST genes along with the development and aging of C. sinensis, and against various exogenous chemical stimuli strongly suggest that the paralogous sigma GST genes might be undergone specialized evolution to cope with the diverse hostile biochemical environments within the mammalian hepatobiliary ductal system. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Maize w3 disrupts homogentisate solanesyl transferase (ZmHst) and reveals a plastoquinone-9 independent path for phytoene desaturation and tocopherol accumulation in kernels

    Science.gov (United States)

    Maize white seedling 3 (w3) has been used to study carotenoid deficiency for almost 100 years, although its genetic basis remained unknown. We show here that w3 phenotype is caused by disruption of homogentisate solanesyl transferase (HST), which catalyzes the first committed step in plastoquinone-9...

  12. Succinyl-CoA:acetoacetate transferase deficiency : identification of a new patient with a neonatal onset and review of the literature

    NARCIS (Netherlands)

    Niezen-Koning, K E; Wanders, R J; Ruiter, J P; Ijlst, L; Visser, G; Reitsma-Bierens, W C; Heijmans, Hugo; Reijngoud, D J; Smit, G P

    1997-01-01

    UNLABELLED: We describe the clinical symptoms and biochemical findings of a patient with succinyl-CoA:acetoacetate transferase deficiency who presented in the neonatal period and review the current literature on this subject. Our patient was initially suspected to have distal renal tubular acidosis,

  13. Effects of Deletion of the Streptococcus pneumoniae Lipoprotein Diacylglyceryl Transferase Gene lgt on ABC Transporter Function and on Growth In Vivo.

    NARCIS (Netherlands)

    Chimalapati, S.; Cohen, J.M.; Camberlein, E.; Macdonald, N.; Durmort, C.; Vernet, T.; Hermans, P.W.M.; Mitchell, T.; Brown, J.S.

    2012-01-01

    Lipoproteins are an important class of surface associated proteins that have diverse roles and frequently are involved in the virulence of bacterial pathogens. As prolipoproteins are attached to the cell membrane by a single enzyme, prolipoprotein diacylglyceryl transferase (Lgt), deletion of the

  14. Succinyl-CoA:acetoacetate transferase deficiency: identification of a new patient with a neonatal onset and review of the literature

    NARCIS (Netherlands)

    Niezen-Koning, K. E.; Wanders, R. J.; Ruiter, J. P.; IJlst, L.; Visser, G.; Reitsma-Bierens, W. C.; Heymans, H. S.; Reijngoud, D. J.; Smit, G. P.

    1997-01-01

    We describe the clinical symptoms and biochemical findings of a patient with succinyl-CoA:acetoacetate transferase deficiency who presented in the neonatal period and review the current literature on this subject. Our patient was initially suspected to have distal renal tubular acidosis, and

  15. The interaction of glutathione S-transferase M1-null variants with tobacco smoke exposure and the development of childhood asthma

    DEFF Research Database (Denmark)

    Rogers, A J; Brasch-Andersen, C; Ionita-Laza, I

    2009-01-01

    BACKGROUND: The glutathione S-transferase M1 (GSTM1)-null variant is a common copy number variant associated with adverse pulmonary outcomes, including asthma and airflow obstruction, with evidence of important gene-by-environment interactions with exposures to oxidative stress. OBJECTIVE: To exp...

  16. Quantum Mechanical/Molecular Mechanical Free Energy Simulations of the Glutathione S-Transferase (M1-1) Reaction with Phenanthrene 9,10-Oxide

    NARCIS (Netherlands)

    Ridder, L.; Rietjens, I.M.C.M.; Vervoort, J.J.M.; Mulholland, A.J.

    2002-01-01

    Glutathione S-transferases (GSTs) play an important role in the detoxification of xenobiotics in mammals. They catalyze the conjugation of glutathione to a wide range of electrophilic compounds. Phenanthrene 9,10-oxide is a model substrate for GSTs, representing an important group of epoxide

  17. Succinyl-CoA:acetoacetate transferase deficiency : identification of a new patient with a neonatal onset and review of the literature

    NARCIS (Netherlands)

    Niezen-Koning, K E; Wanders, R J; Ruiter, J P; Ijlst, L; Visser, G; Reitsma-Bierens, W C; Heijmans, Hugo; Reijngoud, D J; Smit, G P

    UNLABELLED: We describe the clinical symptoms and biochemical findings of a patient with succinyl-CoA:acetoacetate transferase deficiency who presented in the neonatal period and review the current literature on this subject. Our patient was initially suspected to have distal renal tubular acidosis,

  18. The role of the glutathione S-transferase genes GSTT1, GSTM1, and GSTP1 in acetaminophen-poisoned patients

    DEFF Research Database (Denmark)

    Buchard, Anders; Eefsen, Martin; Semb, Synne

    2012-01-01

    The aim of this study was to assess if genetic variants in the glutathione-S-transferase genes GST-T1, M1, and P1 reflect risk factors in acetaminophen (APAP)-poisoned patients assessed by investigation of the relation to prothrombin time (PT), which is a sensitive marker of survival in these pat...

  19. The association of glutathione S-transferase GSTT1 and GSTM1 gene polymorphism with pseudoexfoliative glaucoma in a Pakistani population.

    NARCIS (Netherlands)

    Khan, M.I.; Micheal, S.; Akhtar, F.; Ahmed, W.; Ijaz, B.; Ahmed, A.; Qamar, R.

    2010-01-01

    PURPOSE: The aim of the present study was to investigate the association of glutathione S-transferase GSTT1 and GSTM1 genotypes with pseudoexfoliative glaucoma (PEXG) in a group of Pakistani patients. METHODS: Multiplex polymerase chain reaction was used to study the GSTT1 and GSTM1 polymorphisms in

  20. Differential transcription of cytochrome P450s and glutathione S transferases in DDT-susceptible and resistant Drosophila melanogaster strains in response to DDT and oxidative stress

    Science.gov (United States)

    Metabolic DDT resistance in Drosophila melanogaster has previously been associated with constitutive over-transcription of cytochrome P450s. Increased P450 activity has also been associated with increased oxidative stress. In contrast, over-transcription of glutathione S transferases (GSTs) has been...

  1. The Effects of Arbuscular Mycorrhiza Fungi on Dry Matter and Concentrations of Nitrogen, Phosphorus and Potassium in Berseem Clover, by Cadmium stress

    Directory of Open Access Journals (Sweden)

    hashem aram

    2016-02-01

    Zanjan, after the complete analysis of soil and obtaining the chemical and physical properties in the laboratory. 6 kg of soil was weighed for each pot and then the soil was contaminated. Cadmium sulfate was used in this experiment. The mycorrhizal fungi weighed 150 grams and was mixed with the soil. After mixing the soil with mycorrhizal fungi, the soil was put in pots and then it was cultivated with clover. In this study, clover seeds weighed 0/5 grams and were disinfected with 10% hydrogen peroxide solution and were added to each pot. Distilled water was used for irrigation. After the completion of growth of plants (about 70 day, plant aerial parts and roots were harvested and before measuring, they were washed with distilled water and then were dried in the oven for 72 hours. Plant aerial parts were harvested. Data were analyzed by SAS (version 9 and MSTATC (version 2.10 software, and obtained variance analysis tables. Mean comparison of different treatments was conducted by Duncan test. Charts were obtained by excel software. Results and Discussion: The results showed that the effects of arbuscular mycorrhizal fungi were significant on all traits measured (P< 0.01. With increasing cadmium concentration in soil, dry matter of 37% and 39%, nitrogen concentration of 35% and 28%, Potassium 9/27% and 37%, and phosphorus concentration of 37% and 39%, reduced in root and aerial, respectively. Also the results showed that arbuscular mycorrhizal fungi increased dry matter amount by 42% and 26%, nitrogen concentration by 40.3% and 30%, phosphorus concentration by 6% and 15.4%, potassium concentrations by 54% and 91.2% in root and aerial, respectively. Interaction between cadmium levels and mycorrhizal fungi in statistics was significant on dry matter aerial, nitrogen concentration in aerial and root, and potassium concentrations in plant root (P< 0.01. Conclusion: The results showed that mycorrhizal fungi were significant on all traits measured in one percent level. Cadmium

  2. Elevated gamma glutamyl transferase levels are associated with the location of acute pulmonary embolism. Cross-sectional evaluation in hospital setting

    Directory of Open Access Journals (Sweden)

    Ozge Korkmaz

    Full Text Available ABSTRACT CONTEXT AND OBJECTIVE: The location of embolism is associated with clinical findings and disease severity in cases of acute pulmonary embolism. The level of gamma-glutamyl transferase increases under oxidative stress-related conditions. In this study, we investigated whether gamma-glutamyl transferase levels could predict the location of pulmonary embolism. DESIGN AND SETTING: Hospital-based cross-sectional study at Cumhuriyet University, Sivas, Turkey. METHODS : 120 patients who were diagnosed with acute pulmonary embolism through computed tomography-assisted pulmonary angiography were evaluated. They were divided into two main groups (proximally and distally located, and subsequently into subgroups according to thrombus localization as follows: first group (thrombus in main pulmonary artery; n = 9; second group (thrombus in main pulmonary artery branches; n = 71; third group (thrombus in pulmonary artery segmental branches; n = 34; and fourth group (thrombus in pulmonary artery subsegmental branches; n = 8. RESULTS : Gamma-glutamyl transferase levels on admission, heart rate, oxygen saturation, right ventricular dilatation/hypokinesia, pulmonary artery systolic pressure and cardiopulmonary resuscitation requirement showed prognostic significance in univariate analysis. The multivariate logistic regression model showed that gamma-glutamyl transferase level on admission (odds ratio, OR = 1.044; 95% confidence interval, CI: 1.011-1.079; P = 0.009 and pulmonary artery systolic pressure (OR = 1.063; 95% CI: 1.005-1.124; P = 0.033 remained independently associated with proximally localized thrombus in pulmonary artery. CONCLUSIONS : The findings revealed a significant association between increased existing embolism load in the pulmonary artery and increased serum gamma-glutamyl transferase levels.

  3. Sweet clover production and agronomy.

    Science.gov (United States)

    Goplen, B P

    1980-05-01

    Sweetclover has a notorious reputation for causing "sweetclover disease" when improperly cured. In spite of this, however, sweetclover remains a useful forage legume with valuable agronomic traits. It is drought-resistant and well adapted to Western Canada. Sweetclover is the highest yielding legume forage in this region and is valuable in soil improvement, silage, hay and pasture production and a prized crop for the honey producer. It is the most saline-tolerant of the legumes and is particularly useful on saline "white alkali" soils where cereals and other crops cannot grow. Special precautions are necessary to avoid spoilage and concomitant dicoumarol formation in preserving sweetclover hay and silage. Feeding recommendations are suggested for the safe utilization of spoiled forage. Low coumarin cultivars of sweetclover are completely safe and will not result in sweetclover disease despite spoilage. The breeding program at Saskatoon is expected to produce a new low coumarin (yellow flowered) sweetclover cultivar within the next two years.

  4. P3N-PIPO, a Frameshift Product from the P3 Gene, Pleiotropically Determines the Virulence of Clover Yellow Vein Virus in both Resistant and Susceptible Peas

    Science.gov (United States)

    Suzuki, Haruka; Miyashita, Yuri; Choi, Sun Hee; Hisa, Yusuke; Rihei, Shunsuke; Shimada, Ryoko; Jeon, Eun Jin; Abe, Junya; Uyeda, Ichiro

    2016-01-01

    ABSTRACT Peas carrying the cyv1 recessive resistance gene are resistant to clover yellow vein virus (ClYVV) isolates No.30 (Cl-No.30) and 90-1 (Cl-90-1) but can be infected by a derivative of Cl-90-1 (Cl-90-1 Br2). The main determinant for the breaking of cyv1 resistance by Cl-90-1 Br2 is P3N-PIPO produced from the P3 gene via transcriptional slippage, and the higher level of P3N-PIPO produced by Cl-90-1 Br2 than by Cl-No.30 contributes to the breaking of resistance. Here we show that P3N-PIPO is also a major virulence determinant in susceptible peas that possess another resistance gene, Cyn1, which does not inhibit systemic infection with ClYVV but causes hypersensitive reaction-like lethal systemic cell death. We previously assumed that the susceptible pea cultivar PI 226564 has a weak allele of Cyn1. Cl-No.30 did not induce cell death, but Cl-90-1 Br2 killed the plants. Our results suggest that P3N-PIPO is recognized by Cyn1 and induces cell death. Unexpectedly, heterologously strongly expressed P3N-PIPO of Cl-No.30 appears to be recognized by Cyn1 in PI 226564. The level of P3N-PIPO accumulation from the P3 gene of Cl-No.30 was significantly lower than that of Cl-90-1 Br2 in a Nicotiana benthamiana transient assay. Therefore, Cyn1-mediated cell death also appears to be determined by the level of P3N-PIPO. The more efficiently a ClYVV isolate broke cyv1 resistance, the more it induced cell death systemically (resulting in a loss of the environment for virus accumulation) in susceptible peas carrying Cyn1, suggesting that antagonistic pleiotropy of P3N-PIPO controls the resistance breaking of ClYVV. IMPORTANCE Control of plant viral disease has relied on the use of resistant cultivars; however, emerging mutant viruses have broken many types of resistance. Recently, we revealed that Cl-90-1 Br2 breaks the recessive resistance conferred by cyv1, mainly by accumulating a higher level of P3N-PIPO than that of the nonbreaking isolate Cl-No.30. Here we show that a

  5. Antioxidant role of glutathione S-transferases: 4-Hydroxynonenal, a key molecule in stress-mediated signaling

    International Nuclear Information System (INIS)

    Singhal, Sharad S.; Singh, Sharda P.; Singhal, Preeti; Horne, David; Singhal, Jyotsana; Awasthi, Sanjay

    2015-01-01

    4-Hydroxy-2-trans-nonenal (4HNE), one of the major end products of lipid peroxidation (LPO), has been shown to induce apoptosis in a variety of cell lines. It appears to modulate signaling processes in more than one way because it has been suggested to have a role in signaling for differentiation and proliferation. It has been known that glutathione S-transferases (GSTs) can reduce lipid hydroperoxides through their Se-independent glutathione-peroxidase activity and that these enzymes can also detoxify LPO end-products such as 4HNE. Available evidence from earlier studies together with results of recent studies in our laboratories strongly suggests that LPO products, particularly hydroperoxides and 4HNE, are involved in the mechanisms of stress-mediated signaling and that it can be modulated by the alpha-class GSTs through the regulation of the intracellular concentrations of 4HNE. We demonstrate that 4HNE induced apoptosis in various cell lines is accompanied with c-Jun-N-terminal kinase (JNK) and caspase-3 activation. Cells exposed to mild, transient heat or oxidative stress acquire the capacity to exclude intracellular 4HNE at a faster rate by inducing GSTA4-4 which conjugates 4HNE to glutathione (GSH), and RLIP76 which mediates the ATP-dependent transport of the GSH-conjugate of 4HNE (GS-HNE). The balance between formation and exclusion promotes different cellular processes — higher concentrations of 4HNE promote apoptosis; whereas, lower concentrations promote proliferation. In this article, we provide a brief summary of the cellular effects of 4HNE, followed by a review of its GST-catalyzed detoxification, with an emphasis on the structural attributes that play an important role in the interactions with alpha-class GSTA4-4. Taken together, 4HNE is a key signaling molecule and that GSTs being determinants of its intracellular concentrations, can regulate stress-mediated signaling, are reviewed in this article. - Highlights: • GSTs are the major

  6. Relating N2O emissions from energy crops to the avoided fossil fuel-derived CO2 – a study on bioethanol and biogas produced from organically managed maize, rye, vetch and grass-clover

    DEFF Research Database (Denmark)

    Carter, Mette Sustmann; Hauggard-Nielsen, Henrik; Thomsen, Sune Tjalfe

    2010-01-01

    ‐derived CO2, where the N2O emission has been subtracted. This value does not account for farm machinery CO2 emissions and fuel consumption during biofuel production. We obtained the greatest net reduction in greenhouse gas emissions by co‐production of bioethanol and biogas or by biogas alone produced from...... fuel‐derived CO2, which is obtained when energy crops are used for biofuel production. The analysis includes five organically managed crops (viz. maize, rye, rye‐vetch, vetch and grass‐clover) and three scenarios for conversion of biomass to biofuel. The scenarios are 1) bioethanol production, 2......) biogas production and 3) co‐production of bioethanol and biogas, where the energy crops are first used for bioethanol fermentation and subsequently the residues from this process are utilized for biogas production. The net reduction in greenhouse gas missions is calculated as the avoided fossil fuel...

  7. Determination of region-specific data of yield and quality of alternatives to silage maize in fodder crops – field trails with forage gras and clover grass mixtures, Sorghum as well as whole plant silage of grain

    Directory of Open Access Journals (Sweden)

    Wosnitza, Andrea

    2014-02-01

    Full Text Available This project should generate current regional results over a period of three years about the parameter yield and quality of alternative fodder crops to maize; this includes grass and clover grass mixtures, silage maize, varieties of Sorghum/millets and whole plant silages of wheat, rye and triticale. The tested silage maize showed the highest and most reliable average dry matter yield with 23 tons per hectare, with a very low variance. The Sorghum and millet varieties had dry matter yields of 3 to 5 tons per hectare below the silage maize yield but with individual values fluctuating in a broad range within years and locations. With values far below 28% the dry matter contents were not suitable for ensiling. The grass and clover grass mixtures are good, stable and established alternatives to maize for silage. They achieved high yields comparable with these of Sorghum but stable and with a highly suitable dry matter content for ensiling. The yield of the whole plant silages was up to 22% lower compared with maize. So none of the alternative crops can compete with the high level yield of silage maize in its favoured region, therefore would be a combination of two crops recommended. But some individual locally adapted mixtures or varieties of the alternative crops reached nearly 80% of the maize yield. Silage maize showed the highest level of the net energy content for lactation (NEL, followed by the values of the fodder crops and the whole plant silages. The Sorghum varieties showed the lowest NEL value of all tested cultures. The highest crude protein showed the fodder crops contents. Silage maize, Sorghum and the whole plant silages had values lying nearly around the 50% mark of the fodder crops.

  8. THE EFFECT OF POLYMORPHISM IN GLUTATHIONE S-TRANSFERASES ON THE DEVELOPING SECOND MALIGNANT NEOPLASMS AFTER LEUKEMIA TREATMENT IN CHILDHOOD

    Directory of Open Access Journals (Sweden)

    Janez Jazbec

    2004-12-01

    Full Text Available Background. Survivors of childhood leukemia have an increased risk of developing second malignant neoplasms and specific treatment factors such as alkylating agents, topoisomerase inhibitors and radiation have been associated with their occurrence. Genetic polymorphism in drug-metabolizing enzymes may result in impared detoxification of chemotherapeutics and may lead to increased risk for cancer.Methods. To test if polymorphism in glutathione S-transferases (GST genes is associated with occurrence of secondary malignant neoplasms, we compared GSTM1, GSTT1 and GSTP1 genotypes among 16 patients treated for childhood leukemia in whom second neoplasm occurred and matched the control group.Results. GSTM1 null genotype was found in 44% of patients with second neoplasms and in 50% in control group (p = 0.768, GSTT1 null genotype in 19% of cases and in 29% of controls (p = 0.729 and GSTP1 105 Ile/ile in 50% of cases and 37% of controls (p = 0.537. Differences in distribution of GST genotypes in patients with second neoplasms after childhood leukemia, compared to a matched control group of patients were not statistically significant.Conclusions. In our study we were not able to show relation between GST genotype and occurrence of second neoplasms after the childhood acute leukemia.

  9. Noninvasive evaluation of adult onset myopathy from carnitine palmitoyl transferase II deficiency using proton magnetic resonance spectroscopy.

    Science.gov (United States)

    Videen, J S; Haseler, L J; Karpinski, N C; Terkeltaub, R A

    1999-08-01

    The adult onset metabolic myopathy of carnitine palmitoyl transferase II (CPT II) deficiency is under-recognized, in part due to variable degrees of enzyme deficiency and symptomatology, as well as limitations in means for noninvasive evaluation. We describe a proton magnetic resonance spectroscopy (MRS) technique, using a standard clinical magnetic resonance imaging scanner, to diagnose and help monitor the response to therapy in adult CPT II deficiency. A 53-year-old woman presented with a long standing history of diffuse aching and fatigue provoked by high fat intake, fasting, or prolonged exertion. Muscle biopsy revealed myopathic features and a deficiency (33% of control) of CPT II activity with elevated palmitoyl carnitine. Proton MRS of the soleus muscle was performed using a 1.5 Tesla scanner before and during dietary therapy. Proton MRS revealed shortening of the transverse relaxation time (T2), consistent with increased acetylation of the carnitine pool. The symptoms resolved completely by treatment with frequent feedings of a high carbohydrate diet low in long chain fatty acids supplemented with medium chain triglycerides and L-carnitine. Recovery of normal muscle MRS and carnitine T2 relaxation was documented by the third month of therapy. Proton MRS is a novel, potentially useful, and readily available adjunct in the diagnosis and therapeutic monitoring of muscle CPT II deficiency.

  10. Purification and properties of an O-acetyl-transferase from Escherichia coli that can O-acetylate polysialic acid sequences

    International Nuclear Information System (INIS)

    Higa, H.; Varki, A.

    1986-01-01

    Certain strains of bacteria synthesize an outer polysialic acid (K1) capsule. Some strains of K1 + E.coli are also capable of adding O-acetyl-esters to the exocyclic hydroxyl groups of the sialic acid residues. Both the capsule and the O-acetyl modification have been correlated with differences in antigenicity and pathogenicity. The authors have developed an assay for an O-acetyl-transferase in E.coli that transfers O-[ 3 H]acetyl groups from [ 3 H]acetyl-Coenzyme A to colominic acid (fragments of the polysialic acid capsule). Using this assay, the enzyme was solubilized, and purified ∼ 600-fold using a single affinity chromatography step with Procion Red-A Agarose. The enzyme also binds to Coenzyme A Sepharose, and can be eluted with high salt or Coenzyme A. The partially purified enzyme has a pH optimum of 7.0 - 7.5, is unaffected by divalent cations, is inhibited by high salt concentrations, is inhibited by Coenzyme A (50% inhibition at 100 μM), and shows an apparent Km for colominic acid of 3.7 mM (sialic acid concentration). This enzyme could be involved in the O-acetyl +/- form variation seen in some strains of K1 + E.coli

  11. Differential substrate behaviours of ethylene oxide and propylene oxide towards human glutathione transferase theta hGSTT1-1.

    Science.gov (United States)

    Thier, R; Wiebel, F A; Bolt, H M

    1999-11-01

    The transformation of ethylene oxide (EO), propylene oxide (PO) and 1-butylene oxide (1-BuO) by human glutathione transferase theta (hGSTT1-1) was studied comparatively using 'conjugator' (GSTT1 + individuals) erythrocyte lysates. The relative sequence of velocity of enzymic transformation was PO > EO > 1-BuO. The faster transformation of PO compared to EO was corroborated in studies with human and rat GSTT1-1 (hGSTT1-1 and rGSTT1-1, respectively) expressed by Salmonella typhimurium TA1535. This sequence of reactivities of homologous epoxides towards GSTT1-1 contrasts to the sequence observed in homologous alkyl halides (methyl bromide, MBr; ethyl bromide, EtBr; n-propyl bromide, PrBr) where the relative sequence MeBr > EtBr > PrBr is observed. The higher reactivity towards GSTT1-1 of propylene oxide compared to ethylene oxide is consistent with a higher chemical reactivity. This is corroborated by experimental data of acid-catalysed hydrolysis of a number of aliphatic epoxides, including ethylene oxide and propylene oxide and consistent with semi-empirical molecular orbital modelings.

  12. A Turkish Patient With Succinyl-CoA:3-Oxoacid CoA Transferase Deficiency Mimicking Diabetic Ketoacidosis

    Directory of Open Access Journals (Sweden)

    Sahin Erdol MD

    2016-05-01

    Full Text Available Succinyl-CoA:3-oxoacid CoA transferase (SCOT deficiency is an autosomal recessive disorder of ketone body utilization that is clinically characterized with intermittent ketoacidosis crises. We report here the second Turkish case with SCOT deficiency. She experienced 3 ketoacidotic episodes: The first ketoacidotic crisis mimicked diabetic ketoacidosis because of the associated hyperglycemia. Among patients with SCOT deficiency, the blood glucose levels at the first crises were variable, and this case had the highest ever reported blood glucose level. She is a compound heterozygote with 2 novel mutations, c.517A>G (K173E and c.1543A>G (M515V, in exons 5 and 17 of the OXCT1 gene, respectively. In patient’s fibroblasts, SCOT activity was deficient and, by immunoblot analysis, SCOT protein was much reduced. The patient attained normal development and had no permanent ketosis. The accurate diagnosis of SCOT deficiency in this case had a vital impact on the management strategy and outcome.

  13. Fractal binding and dissociation kinetics of lecithin cholesterol acyl transferase (LCAT), a heart-related compound, on biosensor surfaces

    Science.gov (United States)

    Doke, Atul M.; Sadana, Ajit

    2006-05-01

    A fractal analysis is presented for the binding and dissociation of different heart-related compounds in solution to receptors immobilized on biosensor surfaces. The data analyzed include LCAT (lecithin cholesterol acyl transferase) concentrations in solution to egg-white apoA-I rHDL immobilized on a biosensor chip surface.1 Single- and dual- fractal models were employed to fit the data. Values of the binding and the dissociation rate coefficient(s), affinity values, and the fractal dimensions were obtained from the regression analysis provided by Corel Quattro Pro 8.0 (Corel Corporation Limited).2 The binding rate coefficients are quite sensitive to the degree of heterogeneity on the sensor chip surface. Predictive equations are developed for the binding rate coefficient as a function of the degree of heterogeneity present on the sensor chip surface and on the LCAT concentration in solution, and for the affinity as a function of the ratio of fractal dimensions present in the binding and the dissociation phases. The analysis presented provided physical insights into these analyte-receptor reactions occurring on different biosensor surfaces.

  14. Proteomic Profiling of Cytosolic Glutathione Transferases from Three Bivalve Species: Corbicula fluminea, Mytilus galloprovincialis and Anodonta cygnea

    Directory of Open Access Journals (Sweden)

    José Carlos Martins

    2014-01-01

    Full Text Available Suspension-feeding bivalves are considered efficient toxin vectors with a relative insensitivity to toxicants compared to other aquatic organisms. This fact highlights the potential role of detoxification enzymes, such as glutathione transferases (GSTs, in this bivalve resistance. Nevertheless, the GST system has not been extensively described in these organisms. In the present study, cytosolic GSTs isoforms (cGST were surveyed in three bivalves with different habitats and life strategies: Corbicula fluminea, Anodonta cygnea and Mytilus galloprovincialis. GSTs were purified by glutathione-agarose affinity chromatography, and the collection of expressed cGST classes of each bivalve were identified using a proteomic approach. All the purified extracts were also characterized kinetically. Results reveal variations in cGST subunits collection (diversity and properties between the three tested bivalves. Using proteomics, four pi-class and two sigma-class GST subunits were identified in M. galloprovincialis. C. fluminea also yielded four pi-class and one sigma-class GST subunits. For A. cygnea, two mu-class and one pi-class GST subunits were identified, these being the first record of GSTs from these freshwater mussels. The affinity purified extracts also show differences regarding enzymatic behavior among species. The variations found in cGST collection and kinetics might justify diverse selective advantages for each bivalve organism.

  15. In silico genome-wide identification and characterization of the glutathione S-transferase gene family in Vigna radiata.

    Science.gov (United States)

    Vaish, Swati; Awasthi, Praveen; Tiwari, Siddharth; Tiwari, Shailesh Kumar; Gupta, Divya; Basantani, Mahesh Kumar

    2018-05-01

    Plant glutathione S-transferases (GSTs) are integral to normal plant metabolism and biotic and abiotic stress tolerance. The GST gene family has been characterized in diverse plant species using molecular biology and bioinformatics approaches. In the current study, in silico analysis identified 44 GSTs in Vigna radiata. Of the total 44 GSTs identified, chromosomal locations of 31 GSTs were confirmed. The pI value of GST proteins ranged from 5.10 to 9.40. The predicted molecular weights ranged from 13.12 to 50 kDa. Subcellular localization analysis revealed that all GSTs were predominantly localized in the cytoplasm. The active site amino acids were confirmed to be serine in tau, phi, theta, zeta, and TCHQD; cysteine in lambda, DHAR, and omega; and tyrosine in EF1G. The gene architecture conformed to the two-exon/one-intron and three-exon/two-intron organization in the case of tau and phi classes, respectively. MEME analysis identified 10 significantly conserved motifs with the width of 8-50 amino acids. The motifs identified were either specific to a specific GST class or were shared by multiple GST classes. The results of the current study will be of potential importance in the characterization of the GST gene family in V. radiata, an economically important leguminous crop.

  16. Crystal Structure of Saccharomyces cerevisiae ECM4, a Xi-Class Glutathione Transferase that Reacts with Glutathionyl-(hydro)quinones

    Science.gov (United States)

    Schwartz, Mathieu; Didierjean, Claude; Hecker, Arnaud; Girardet, Jean-Michel; Morel-Rouhier, Mélanie; Gelhaye, Eric; Favier, Frédérique

    2016-01-01

    Glutathionyl-hydroquinone reductases (GHRs) belong to the recently characterized Xi-class of glutathione transferases (GSTXs) according to unique structural properties and are present in all but animal kingdoms. The GHR ScECM4 from the yeast Saccharomyces cerevisiae has been studied since 1997 when it was found to be potentially involved in cell-wall biosynthesis. Up to now and in spite of biological studies made on this enzyme, its physiological role remains challenging. The work here reports its crystallographic study. In addition to exhibiting the general GSTX structural features, ScECM4 shows extensions including a huge loop which contributes to the quaternary assembly. These structural extensions are probably specific to Saccharomycetaceae. Soaking of ScECM4 crystals with GS-menadione results in a structure where glutathione forms a mixed disulfide bond with the cysteine 46. Solution studies confirm that ScECM4 has reductase activity for GS-menadione in presence of glutathione. Moreover, the high resolution structures allowed us to propose new roles of conserved residues of the active site to assist the cysteine 46 during the catalytic act. PMID:27736955

  17. Crystal Structure of Saccharomyces cerevisiae ECM4, a Xi-Class Glutathione Transferase that Reacts with Glutathionyl-(hydroquinones.

    Directory of Open Access Journals (Sweden)

    Mathieu Schwartz

    Full Text Available Glutathionyl-hydroquinone reductases (GHRs belong to the recently characterized Xi-class of glutathione transferases (GSTXs according to unique structural properties and are present in all but animal kingdoms. The GHR ScECM4 from the yeast Saccharomyces cerevisiae has been studied since 1997 when it was found to be potentially involved in cell-wall biosynthesis. Up to now and in spite of biological studies made on this enzyme, its physiological role remains challenging. The work here reports its crystallographic study. In addition to exhibiting the general GSTX structural features, ScECM4 shows extensions including a huge loop which contributes to the quaternary assembly. These structural extensions are probably specific to Saccharomycetaceae. Soaking of ScECM4 crystals with GS-menadione results in a structure where glutathione forms a mixed disulfide bond with the cysteine 46. Solution studies confirm that ScECM4 has reductase activity for GS-menadione in presence of glutathione. Moreover, the high resolution structures allowed us to propose new roles of conserved residues of the active site to assist the cysteine 46 during the catalytic act.

  18. Inhibition of carnitine-acyl transferase I by oxfenicine studied in vivo with [11C]-labeled fatty acids

    International Nuclear Information System (INIS)

    Angsten, Gertrud; Valind, Sven; Takalo, Reijo; Neu, Henrik; Meurling, Staffan; Langstroem, Bengt

    2005-01-01

    Methods: Anesthetized pigs were studied with [ 11 C]-labeled fatty acids (FAs) with carbon chain length ranging from 8 to 16 carbon atoms, during control conditions and during inhibition of carnitine-palmitoyl transferase I (CPT I) with oxfenicine. The myocardial uptake of [ 11 C]-FAs from blood was measured together with the relative distribution of [ 11 C]-acyl-CoA between rapid mitochondrial oxidation and incorporation into slow turnover lipid pools in the heart. Results: During baseline conditions, the fractional oxidative utilization of palmitate was almost as high as that of carnitine-independent short-chain FAs, unless the carnitine shuttle was inhibited by high levels of lactate. Inhibition of CPT I almost completely blocked the oxidative pathway for palmitic acid and reduced the fractional oxidative utilization, while the rate of oxidative metabolism of acyl-CoA was unaffected. Conclusions: [ 11 C]-Labeled FAs allow rapid oxidation to be well separated from esterification into slow turnover lipid pools in the heart of anaesthetized pigs. The fractional oxidative utilization of [ 11 C]-palmitate serves well to characterize, in vivo, the carnitine-dependent transfer of long-chain FAs

  19. Inhibition of carnitine-acyl transferase I by oxfenicine studied in vivo with [{sup 11}C]-labeled fatty acids

    Energy Technology Data Exchange (ETDEWEB)

    Angsten, Gertrud [Department of Pediatric Surgery, University Children' s Hospital, S-751 85 Uppsala (Sweden)]. E-mail: gertrud.angsten@surgsci.uu.se; Valind, Sven [Uppsala University PET Centre, Uppsala University, S-751 05 Uppsala (Sweden); Department of Clinical Physiology, University Hospital, S-751 85 Uppsala (Sweden); Takalo, Reijo [Uppsala University PET Centre, Uppsala University, S-751 05 Uppsala (Sweden); Department of Clinical Physiology, University Hospital, S-751 85 Uppsala (Sweden); Neu, Henrik [Uppsala University PET Centre, Uppsala University, S-751 05 Uppsala (Sweden); Department of Organic Chemistry, Uppsala University, S-751 24 Uppsala (Sweden); Meurling, Staffan [Department of Pediatric Surgery, University Children' s Hospital, S-751 85 Uppsala (Sweden); Langstroem, Bengt [Uppsala University PET Centre, Uppsala University, S-751 05 Uppsala (Sweden); Department of Organic Chemistry, Uppsala University, S-751 24 Uppsala (Sweden)

    2005-07-01

    Methods: Anesthetized pigs were studied with [{sup 11}C]-labeled fatty acids (FAs) with carbon chain length ranging from 8 to 16 carbon atoms, during control conditions and during inhibition of carnitine-palmitoyl transferase I (CPT I) with oxfenicine. The myocardial uptake of [{sup 11}C]-FAs from blood was measured together with the relative distribution of [{sup 11}C]-acyl-CoA between rapid mitochondrial oxidation and incorporation into slow turnover lipid pools in the heart. Results: During baseline conditions, the fractional oxidative utilization of palmitate was almost as high as that of carnitine-independent short-chain FAs, unless the carnitine shuttle was inhibited by high levels of lactate. Inhibition of CPT I almost completely blocked the oxidative pathway for palmitic acid and reduced the fractional oxidative utilization, while the rate of oxidative metabolism of acyl-CoA was unaffected. Conclusions: [{sup 11}C]-Labeled FAs allow rapid oxidation to be well separated from esterification into slow turnover lipid pools in the heart of anaesthetized pigs. The fractional oxidative utilization of [{sup 11}C]-palmitate serves well to characterize, in vivo, the carnitine-dependent transfer of long-chain FAs.

  20. Mechanistic insights into EgGST1, a Mu class glutathione S-transferase from the cestode parasite Echinococcus granulosus.

    Science.gov (United States)

    Arbildi, Paula; Turell, Lucía; López, Verónica; Alvarez, Beatriz; Fernández, Verónica

    2017-11-01

    Glutathione transferases (GSTs) comprise a major detoxification system in helminth parasites, displaying both catalytic and non-catalytic activities. The kinetic mechanism of these enzymes is complex and depends on the isoenzyme which is being analyzed. Here, we characterized the kinetic mechanism of rEgGST1, a recombinant form of a cytosolic GST from Echinococcus granulosus (EgGST1), which is related to the Mu-class of mammalian enzymes, using the canonical substrates glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). Initial rate and product inhibition studies were consistent with a steady-state random sequential mechanism, where both substrates are bound to the enzyme before the products are released. Kinetic constants were also determined (pH 6.5 and 30 °C). Moreover, rEgGST1 lowered the pK a of GSH from 8.71 ± 0.07 to 6.77 ± 0.08, and enzyme-bound GSH reacted with CDNB 1 × 10 5 times faster than free GSH at pH 7.4. Finally, the dissociation of the enzyme-GSH complex was studied by means of intrinsic fluorescence, as well as that of the complex with the anthelminth drug mebendazole. This is the first report on mechanistic issues related to a helminth parasitic GST. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Human T cell recognition of the blood stage antigen Plasmodium hypoxanthine guanine xanthine phosphoribosyl transferase (HGXPRT in acute malaria

    Directory of Open Access Journals (Sweden)

    Woodberry Tonia

    2009-06-01

    Full Text Available Abstract Background The Plasmodium purine salvage enzyme, hypoxanthine guanine xanthine phosphoribosyl transferase (HGXPRT can protect mice against Plasmodium yoelii pRBC challenge in a T cell-dependent manner and has, therefore, been proposed as a novel vaccine candidate. It is not known whether natural exposure to Plasmodium falciparum stimulates HGXPRT T cell reactivity in humans. Methods PBMC and plasma collected from malaria-exposed Indonesians during infection and 7–28 days after anti-malarial therapy, were assessed for HGXPRT recognition using CFSE proliferation, IFNγ ELISPOT assay and ELISA. Results HGXPRT-specific T cell proliferation was found in 44% of patients during acute infection; in 80% of responders both CD4+ and CD8+ T cell subsets proliferated. Antigen-specific T cell proliferation was largely lost within 28 days of parasite clearance. HGXPRT-specific IFN-γ production was more frequent 28 days after treatment than during acute infection. HGXPRT-specific plasma IgG was undetectable even in individuals exposed to malaria for at least two years. Conclusion The prevalence of acute proliferative and convalescent IFNγ responses to HGXPRT demonstrates cellular immunogenicity in humans. Further studies to determine minimal HGXPRT epitopes, the specificity of responses for Plasmodia and associations with protection are required. Frequent and robust T cell proliferation, high sequence conservation among Plasmodium species and absent IgG responses distinguish HGXPRT from other malaria antigens.

  2. γ-Glutamyl Transferase as a Risk Factor for All-Cause or Cardiovascular Disease Mortality Among 5912 Ischemic Stroke.

    Science.gov (United States)

    Tu, Wen-Jun; Liu, Qiang; Cao, Jian-Lei; Zhao, Sheng-Jie; Zeng, Xian-Wei; Deng, Ai-Jun

    2017-10-01

    The aim of the study was to evaluate the association of the measurement of serum γ-glutamyl transferase (GGT) concentrations at admission with 1-year all-cause or cardiovascular disease (CVD) mortality in patients with acute ischemic stroke. This prospective, multicenter cohort study was conducted in 4 stroke centers in China. Baseline GGT measurements were tested. The relationship of GGT to the risk of death from all-cause or CVD was examined among 1-year follow-up patients. We recorded results from 5912 patients with stroke. In those patients, 51.0% were men, and the median age was 61 years. In both men and women, high GGT was significantly associated with total mortality from all-cause or CVD ( P mortality from all-cause and CVD, respectively. With an area under the curve of 0.69 (95% confidence interval, 0.66-0.73), GGT showed a significantly greater discriminatory ability to predict all-cause mortality as compared with others factors. GGT improved the National Institutes of Health Stroke Scale score (area under the curve of the combined model, 0.75 [95% confidence interval, 0.73-0.78]; P mortality in patients with ischemic stroke. © 2017 American Heart Association, Inc.

  3. Glutathione S-transferase of brown planthoppers (Nilaparvata lugens is essential for their adaptation to gramine-containing host plants.

    Directory of Open Access Journals (Sweden)

    Xiao-Qin Sun

    Full Text Available Plants have evolved complex processes to ward off attacks by insects. In parallel, insects have evolved mechanisms to thwart these plant defenses. To gain insight into mechanisms that mediate this arms race between plants and herbivorous insects, we investigated the interactions between gramine, a toxin synthesized by plants of the family Gramineae, and glutathione S transferase (GST, an enzyme found in insects that is known to detoxify xenobiotics. Here, we demonstrate that rice (Oryza sativa, a hydrophytic plant, also produces gramine and that rice resistance to brown planthoppers (Nilaparvata lugens, BPHs is highly associated with in planta gramine content. We also show that gramine is a toxicant that causes BPH mortality in vivo and that knockdown of BPH GST gene nlgst1-1 results in increased sensitivity to diets containing gramine. These results suggest that the knockdown of key detoxification genes in sap-sucking insects may provide an avenue for increasing their sensitivity to natural plant-associated defense mechanisms.

  4. Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT of Plasmodium falciparum

    Directory of Open Access Journals (Sweden)

    Carlos Alfonso Nieto Clavijo

    2017-09-01

    Full Text Available Recombinant proteins have become useful tools in biochemistry research. During their production, however, inclusion bodies (IB appear, on the one hand, due to the high expression rate from the recombinant plasmids, which have high efficiency promoters, and, on the other hand, intrinsic characteristics of the expressed protein. Furhtermore, the nicotinamide/nicotinate mononucleotide adenilyl transferase (NMNAT is a central protein in NAD(H+ biosynthesis, an essential cofactor in cell metabolism, and in protozoon parasite has been studied. To study the NMNAT protein of these parasites, their recombinant version in E. coli has been expressed, getting a great quantity of IB as a by-product. To increase the solubility of the protein, the coding sequence of the NMNAT enzyme of Plasmodium falciparum was cloned in different expression plasmids which were subsequently transformed into E. coli BL21(DE3 expression strain. The solubility of the recombinant proteins was assessed and the one with the highest presence in the soluble fraction was subsequently purified and its enzyme activity was determined. The recombinant protein with a MBP (maltose-binding protein tag showed an increased solubility and purity.

  5. Purification and properties of the glutathione S-transferases from the anoxia-tolerant turtle, Trachemys scripta elegans.

    Science.gov (United States)

    Willmore, William G; Storey, Kenneth B

    2005-07-01

    Glutathione S-transferases (GSTs) play critical roles in detoxification, response to oxidative stress, regeneration of S-thiolated proteins, and catalysis of reactions in nondetoxification metabolic pathways. Liver GSTs were purified from the anoxia-tolerant turtle, Trachemys scripta elegans. Purification separated a homodimeric (subunit relative molecular mass =34 kDa) and a heterodimeric (subunit relative molecular mass = 32.6 and 36.8 kDa) form of GST. The enzymes were purified 23-69-fold and 156-174-fold for homodimeric and heterodimeric GSTs, respectively. Kinetic data gathered using a variety of substrates and inhibitors suggested that both homodimeric and heterodimeric GSTs were of the alpha class although they showed significant differences in substrate affinities and responses to inhibitors. For example, homodimeric GST showed activity with known alpha class substrates, cumene hydroperoxide and p-nitrobenzylchloride, whereas heterodimeric GST showed no activity with cumene hydroperoxide. The specific activity of liver GSTs with chlorodinitrobenzene (CDNB) as the substrate was reduced by 2.6- and 8.7-fold for homodimeric and heterodimeric GSTs isolated from liver of anoxic turtles as compared with aerobic controls, suggesting an anoxia-responsive stable modification of the protein that may alter its function during natural anaerobiosis.

  6. Designing Ligands for Leishmania, Plasmodium, and Aspergillus N-Myristoyl Transferase with Specificity and Anti-Target-Safe Virtual Libraries.

    Science.gov (United States)

    Garcia-Sosa, Alfonso T

    2018-01-01

    Leishmaniasis, malaria, and fungal diseases are burdens on individuals and populations and can present severe complications. Easily accessible chemical treatments for these diseases are increasingly sought-after. Targeting the parasite N-myristoyl transferase while avoiding the human enzyme and other anti-targets may allow the prospect of compounds with pan-activity against these diseases, which would simplify treatments and costs. Developing chemical libraries, both virtual and physical, that have been filtered and flagged early on in the drug discovery process (before virtual screening) could reduce attrition rates of compounds being developed and failing late in development stages due to problems of side-effects or toxicity. Chemical libraries have been screened against the anti-targets pregnane-X-receptor, sulfotransferase, cytochrome P450 2a6, 2c9, and 3a4 with three different docking programs. Statistically significant differences are observed in their interactions with these enzymes as compared to small molecule drugs and bioactive non-drug datasets. A series of compounds are proposed with the best predicted profiles for inhibition of all parasite targets while sparing the human form and anti-targets. Some of the topranked compounds have confirmed experimental activity against Leishmania, and highlighted are those compounds with best properties for further development. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  7. A Conserved Splicing Silencer Dynamically Regulates O-GlcNAc Transferase Intron Retention and O-GlcNAc Homeostasis

    Directory of Open Access Journals (Sweden)

    Sung-Kyun Park

    2017-08-01

    Full Text Available Modification of nucleocytoplasmic proteins with O-GlcNAc regulates a wide variety of cellular processes and has been linked to human diseases. The enzymes O-GlcNAc transferase (OGT and O-GlcNAcase (OGA add and remove O-GlcNAc, but the mechanisms regulating their expression remain unclear. Here, we demonstrate that retention of the fourth intron of OGT is regulated in response to O-GlcNAc levels. We further define a conserved intronic splicing silencer (ISS that is necessary for OGT intron retention. Deletion of the ISS in colon cancer cells leads to increases in OGT, but O-GlcNAc homeostasis is maintained by concomitant increases in OGA protein. However, the ISS-deleted cells are hypersensitive to OGA inhibition in culture and in soft agar. Moreover, growth of xenograft tumors from ISS-deleted cells is compromised in mice treated with an OGA inhibitor. Thus, ISS-mediated regulation of OGT intron retention is a key component in OGT expression and maintaining O-GlcNAc homeostasis.

  8. Volatile Gas Production by Methyl Halide Transferase: An In Situ Reporter Of Microbial Gene Expression In Soil.

    Science.gov (United States)

    Cheng, Hsiao-Ying; Masiello, Caroline A; Bennett, George N; Silberg, Jonathan J

    2016-08-16

    Traditional visual reporters of gene expression have only very limited use in soils because their outputs are challenging to detect through the soil matrix. This severely restricts our ability to study time-dependent microbial gene expression in one of the Earth's largest, most complex habitats. Here we describe an approach to report on dynamic gene expression within a microbial population in a soil under natural water levels (at and below water holding capacity) via production of methyl halides using a methyl halide transferase. As a proof-of-concept application, we couple the expression of this gas reporter to the conjugative transfer of a bacterial plasmid in a soil matrix and show that gas released from the matrix displays a strong correlation with the number of transconjugant bacteria that formed. Gas reporting of gene expression will make possible dynamic studies of natural and engineered microbes within many hard-to-image environmental matrices (soils, sediments, sludge, and biomass) at sample scales exceeding those used for traditional visual reporting.

  9. Association of N-acetyltransferase-2 and glutathione S-transferase polymorphisms with idiopathic male infertility in Vietnam male subjects.

    Science.gov (United States)

    Trang, Nguyen Thi; Huyen, Vu Thi; Tuan, Nguyen Thanh; Phan, Tran Duc

    2018-04-25

    N-acetyltransferase-2 (NAT2) and Glutathione S-transferases (GSTs) are phase-II xenobiotic metabolizing enzymes participating in detoxification of toxic arylamines, aromatic amines, hydrazines and reactive oxygen species (ROS), which are produced under oxidative and electrophile stresses. The purpose of this research was to investigate whether two common single-nucleotide polymorphisms (SNP) of NAT2 (rs1799929, rs1799930) and GSTP1 (rs1138272, rs1695) associated with susceptibility to idiopathic male infertility. A total 300 DNA samples (150 infertile patients and 150 healthy control) were genotyped for the polymorphisms by ARMS - PCR. We revealed a significant association between the NAT2 variant genotypes (CT + TT (rs1799929), (OR: 3.74; p male infertility in subjects from Vietnam. This pilot study is the first (as far as we know) to reveal that polymorphisms of NAT2 (rs1799929, rs1799930) and GSTP1 (rs1138272, rs1695) are some novel genetic markers for susceptibility to idiopathic male infertility. Copyright © 2018 Elsevier B.V. All rights reserved.

  10. Effect of Arsenic and Chromium on the Serum Amino-Transferases Activity in Indian Major Carp, Labeo rohita

    Directory of Open Access Journals (Sweden)

    Anjaneyulu Yerramilli

    2007-09-01

    Full Text Available Arsenic and hexavalent chromium toxicity results from their ability to interact with sulfahydryl groups of proteins and enzymes, and to substitute phosphorus in a variety of biochemical reactions. Alanine aminotransferase (ALT; E.C: 2.6.1.2 and Aspartate amino transferase (AST; EC 2.6.1.1 play a crucial role in transamination reactions and can be used as potential biomarkers to indicate hepatotoxicity and cellular damage. While histopathological studies in liver tissue require more time and expertise, simple and reliable biochemical analysis of ALT and AST can be used for a rapid assessment of tissue and cellular damage within 96 h. The main objective of this study was to determine the acute effects of arsenic and hexavalent chromium on the activity of ALT and AST in the Indian major carp, Labeo rohita for 24 h and 96 h. Significant increase in the activity of ALT (P < 0.01 from controls in arsenic exposed fish indicates serious hepatic damage and distress condition to the fish. However, no such significant changes were observed in chromium-exposed fish suggesting that arsenic is more toxic to the fish. These findings indicate that ALT and AST are candidate biomarkers for arsenic-induced hepatotoxicity in Labeo rohita.

  11. Thymidylate Synthase, Thymidine Phosphorylase and Orotate Phosphoribosyl Transferase Levels as Predictive Factors of Chemotherapy in Oral Squamous Cell Carcinoma

    International Nuclear Information System (INIS)

    Ogiuchi, Yosuke; Maruoka, Yasubumi; Ando, Tomohiro; Kobayashi, Makio; Ogiuchi, Hideki

    2008-01-01

    We conducted a clinicopathologic study on protein and mRNA levels of thymidylate synthase (TS), thymidine phosphorylase (TP) and orotate phosphoribosyl transferase (OPRT) using biopsy tissue specimens before treatment. The mRNA levels have been measured in tumor cells microdissected from paraffin-embedded specimens (Danenberg Tumor Profile method: DTP method). We studied the mRNA and protein expression as effect predictive factors in chemotherapy. The subjects consisted of 20 cases of untreated oral squamous cell carcinoma who had undergone chemotherapy with TS-1 (16 males and 4 females, tongue in 8 cases, upper gingiva in 3 cases, lower gingiva in 3 cases, buccal mucosa in 5 cases and floor of the mouth in 1 case). TS gene expressions of the responders were lower than those for the nonresponders. Furthermore, regarding males who were less than 70 years of age, stage I and II, well differentiated type and tongue, TS mRNA expression of the responders were lower than that for the nonresponders. The mRNA expression of OPRT for the male responders was lower than that for the nonresponders. No remarkable difference was observed by immunohistochemistry. In this study, the measurement of the TS levels using the DTP method may potentially act as a predictive factor of antitumor effectiveness

  12. An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2

    DEFF Research Database (Denmark)

    Zlocowski, Natacha; Lorenz, Virginia; Bennett, Eric Paul

    2013-01-01

    Abstract Polypeptide GalNAc-transferases (ppGalNAc-Ts) are a family of enzymes that catalyze the initiation of mucin-type O-glycosylation. All ppGalNAc-T family members contain a common (QXW)3 motif which is present in R-type lectin group. Acetylation site K521 is part of the QKW motif of ß......-trefoil in the lectin domain of ppGalNAc-T2. We used a combination of acetylation and site-directed mutagenesis approaches to examine the functional role of K521 in ppGalNAc-T2. Binding assays of non-acetylated and acetylated forms of the mutant ppGalNAc-T2K521Q to various naked and aGalNAc-glycosylated mucin peptides...... indicated that degree of interaction of lectin domain with aGalNAc depends on the peptide sequence of mucin. Studies of inhibitory effect of various carbohydrates on interactions of ppGalNAc-T2 with MUC1aGalNAc indicate that point K521Q mutation enhance the carbohydrate specificity of lectin domain for aGalNAc...

  13. Evaluation of the precision-cut liver and lung slice systems for the study of induction of CYP1, epoxide hydrolase and glutathione S-transferase activities.

    Science.gov (United States)

    Pushparajah, Daphnee S; Umachandran, Meera; Plant, Kathryn E; Plant, Nick; Ioannides, Costas

    2007-02-28

    The principal objective was to ascertain whether precision-cut tissue slices can be used to evaluate the potential of chemicals to induce CYP1, epoxide hydrolase and glutathione S-transferase activities, all being important enzymes involved in the metabolism of polycyclic aromatic hydrocarbons. Precision-cut rat liver and lung slices were incubated with a range of benzo[a]pyrene concentrations for various time periods. A rise in the O-deethylation of ethoxyresorufin was seen in both liver and lung slices exposed to benzo[a]pyrene, which was accompanied by increased CYP1A apoprotein levels. Pulmonary CYP1B1 apoprotein levels and hepatic mRNA levels were similarly enhanced. Elevated epoxide hydrolase and glutathione S-transferase activities were also observed in liver slices following incubation for 24h; similarly, a rise in apoprotein levels of both enzymes was evident, peak levels occurring at the same time point. When mRNA levels were monitored, a rise in the levels of both enzymes was seen as early as 4h after incubation, but maximum levels were attained at 24 h. In lung slices, induction of epoxide hydrolase by benzo[a]pyrene was observed after a 24-h incubation, and at a concentration of 1 microM; a rise in apoprotein levels was seen at this time point. Glutathione S-transferase activity was not inducible in lung slices by benzo[a]pyrene but a modest increase was observed in hepatic slices. Collectively, these studies confirmed CYP1A induction in rat liver slices and established that CYP1B1 expression, and epoxide hydrolase and glutathione S-transferase activities are inducible in precision-cut tissue slices.

  14. Purification, molecular cloning and heterologous expression of a glutathione S-transferase involved in insecticide resistance from the rice brown planthopper, Nilaparvata lugens.

    OpenAIRE

    Vontas, John G; Small, Graham J; Nikou, Dimitra C; Ranson, Hilary; Hemingway, Janet

    2002-01-01

    A novel glutathione S-transferase (GST)-based pyrethroid resistance mechanism was recently identified in Nilaparvata lugens [Vontas, Small and Hemingway (2001) Biochem. J. 357, 65-72]. To determine the nature of GSTs involved in conferring this resistance, the GSTs from resistant and susceptible strains of N. lugens were partially purified by anion exchange and affinity chromatography. The majority of peroxidase activity, previously correlated with resistance, was confined to the fraction tha...

  15. An ethylene-responsive enhancer element is involved in the senescence-related expression of the carnation glutathione-S-transferase (GST1) gene.

    OpenAIRE

    Itzhaki, H; Maxson, J M; Woodson, W R

    1994-01-01

    The increased production of ethylene during carnation petal senescence regulates the transcription of the GST1 gene encoding a subunit of glutathione-S-transferase. We have investigated the molecular basis for this ethylene-responsive transcription by examining the cis elements and trans-acting factors involved in the expression of the GST1 gene. Transient expression assays following delivery of GST1 5' flanking DNA fused to a beta-glucuronidase receptor gene were used to functionally define ...

  16. Two pear glutathione S-transferases genes are regulated during fruit development and involved in response to salicylic acid, auxin, and glucose signaling.

    Directory of Open Access Journals (Sweden)

    Hai-Yan Shi

    Full Text Available Two genes encoding putative glutathione S-transferase proteins were isolated from pear (Pyrus pyrifolia and designated PpGST1 and PpGST2. The deduced PpGST1 and PpGST2 proteins contain conserved Glutathione S-transferase N-terminal domain (GST_N and Glutathione S-transferase, C-terminal domain (GST_C. Using PCR amplification technique, the genomic clones corresponding to PpGST1 and PpGST2 were isolated and shown to contain two introns and a singal intron respectively with typical GT/AG boundaries defining the splice junctions. Phylogenetic analysis clearly demonstrated that PpGST1 belonged to Phi class of GST superfamilies and had high homology with apple MdGST, while PpGST2 was classified into the Tau class of GST superfamilies. The expression of PpGST1 and PpGST2 genes was developmentally regulated in fruit. Further study demonstrated that PpGST1 and PpGST2 expression was remarkably induced by glucose, salicylic acid (SA and indole-3-aceticacid (IAA treatments in pear fruit, and in diseased fruit. These data suggested that PpGST1 and PpGST2 might be involved in response to sugar, SA, and IAA signaling during fruit development of pear.

  17. EFEK POLIMORFISME GENA GSTP-1 TERHADAP AKTIVITAS GLUTATION S-TRANSFERASE (GST PADA INDIVIDU TERPAPAR LOGAM BERAT TIMBAL (Effect of GSTP-1 Gene Polymorphismson Glutation S- Transferase (GST Activity in Heavy Metals Lead-Exposed Individual

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    Hernayanti Hernayanti

    2015-11-01

    Full Text Available ABSTRAK Gena GSTP-1 merupakan penghasil enzim glutation S- transferase (GST, yang berfungsi dalam proses detoksifikasi senyawa toksik di hati. Faktor keberadaan polimorfisme gena GSTP-1 akan menyebabkan penurunan ekspresi GST, sehingga proses detoksifikasi terhadap senyawa toksik akan terhambat. Kerentanan terhadap paparan senyawa toksik pada manusia akan meningkat apabila dijumpai polimorfisme gena. Salah satu senyawa toksik yang dapat menghambat aktivitas GST adalah timbal (Pb, terutama dalam bentuk tetra ethyl lead (TEL. Tujuan penelitian adalah untuk mengetahui pengaruh polimorfisme gena GSTP-1 terhadap aktivitas GST pada individu terpapar Pb, yang diwakili pekerja bengkel mobil. Faktor keberadaan polimorfisme gena individu ditentukan dengan metode PCR-RFLP dan enzim restriksi BsmA1. Parameter yang diukur adalah kadar Pb dan aktivitas GST. Analisis molekuler gena GSTP-1 dilakukan secara deskriptif. Data kadar Pb dan aktivitas GST dianalisis dengan uji t independent. Hasil analisis gena GSTP-1 dari 40 orang subyek kasus setelah dilakukan digesti dengan enzim BsmA1, ditemukan sebanyak 10 orang individu dengan polimorfisme Ile105Val gena GSTP 1 atau sekitar 25% dengan genotip Ile-Val, sedangkan 30 orang atau 75% ditemukan tanpa polimorfisme dengan genotip Ile-Ile. Pita DNA individu dengan polimorfisme terpotong menjadi 3 fragmen sepanjang 176, 91 dan 85 pp (mutan heterozygot, sedangkan tanpa polimorfisme terletak pada 176 bp. Subyek kasus dengan polimorfisme gena GSTP-1 memiliki kadar Pb lebih tinggi dan aktivitas GST lebih rendah dibandingkan individu non polimorfisme. Telah terbukti bahwa polimorfisme gena GSTP-1 menyebabkan penurunan ekspresi enzim GST. Pada individu terpapar Pb dengan polimorfisme gena GSTP-1 memiliki aktivitas GST lebih rendah dibandingkan individu tanpa polimorfisme. ABSTRACT GSTP-1 gene regulates the expression of gluthation S-transferase enzyme, which role in detoxification of toxicant on liver. If the polymorphisms

  18. Differential activation of diverse glutathione transferases of Clonorchis sinensis in response to the host bile and oxidative stressors.

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    Young-An Bae

    Full Text Available BACKGROUND: Clonorchis sinensis causes chronic cumulative infections in the human hepatobiliary tract and is intimately associated with cholangiocarcinoma. Approximately 35 million people are infected and 600 million people are at risk of infections worldwide. C. sinensis excretory-secretory products (ESP constitute the first-line effector system affecting the host-parasite interrelationship by interacting with bile fluids and ductal epithelium. However, the secretory behavior of C. sinensis in an environment close to natural host conditions is unclear. C. sinensis differs from Fasciola hepatica in migration to, and maturation in, the hepatic bile duct, implying that protein profile of the ESP of these two trematodes might be different from each other. METHODOLOGY/PRINCIPAL FINDINGS: We conducted systemic approaches to analyze the C. sinensis ESP proteome and the biological reactivity of C. sinensis glutathione transferases (GSTs, such as global expression patterns and induction profiles under oxidative stress and host bile. When we observed ex host excretion behavior of C. sinensis in the presence of 10% host bile, the global proteome pattern was not significantly altered, but the amount of secretory proteins was increased by approximately 3.5-fold. Bioactive molecules secreted by C. sinensis revealed universal/unique features in relation to its intraluminal hydrophobic residing niche. A total of 38 protein spots identified abundantly included enzymes involved in glucose metabolism (11 spots, 28.9% and diverse-classes of glutathione transferases (GSTs; 10 spots, 26.3%. Cathepsin L/F (four spots, 10.5% and transporter molecules (three spots, 7.9% were also recognized. The universal secretory proteins found in other parasites, such as several enzymes involved in glucose metabolism and oxygen transporters, were commonly detected. C. sinensis secreted less cysteine proteases and fatty acid binding proteins compared to other tissue-invading or

  19. Differential Activation of Diverse Glutathione Transferases of Clonorchis sinensis in Response to the Host Bile and Oxidative Stressors

    Science.gov (United States)

    Bae, Young-An; Ahn, Do-Whan; Lee, Eung-Goo; Kim, Seon-Hee; Cai, Guo-Bin; Kang, Insug; Sohn, Woon-Mok; Kong, Yoon

    2013-01-01

    Background Clonorchis sinensis causes chronic cumulative infections in the human hepatobiliary tract and is intimately associated with cholangiocarcinoma. Approximately 35 million people are infected and 600 million people are at risk of infections worldwide. C. sinensis excretory-secretory products (ESP) constitute the first-line effector system affecting the host-parasite interrelationship by interacting with bile fluids and ductal epithelium. However, the secretory behavior of C. sinensis in an environment close to natural host conditions is unclear. C. sinensis differs from Fasciola hepatica in migration to, and maturation in, the hepatic bile duct, implying that protein profile of the ESP of these two trematodes might be different from each other. Methodology/Principal Findings We conducted systemic approaches to analyze the C. sinensis ESP proteome and the biological reactivity of C. sinensis glutathione transferases (GSTs), such as global expression patterns and induction profiles under oxidative stress and host bile. When we observed ex host excretion behavior of C. sinensis in the presence of 10% host bile, the global proteome pattern was not significantly altered, but the amount of secretory proteins was increased by approximately 3.5-fold. Bioactive molecules secreted by C. sinensis revealed universal/unique features in relation to its intraluminal hydrophobic residing niche. A total of 38 protein spots identified abundantly included enzymes involved in glucose metabolism (11 spots, 28.9%) and diverse-classes of glutathione transferases (GSTs; 10 spots, 26.3%). Cathepsin L/F (four spots, 10.5%) and transporter molecules (three spots, 7.9%) were also recognized. The universal secretory proteins found in other parasites, such as several enzymes involved in glucose metabolism and oxygen transporters, were commonly detected. C. sinensis secreted less cysteine proteases and fatty acid binding proteins compared to other tissue-invading or intravascular

  20. Are glutathione S transferases involved in DNA damage signalling? Interactions with DNA damage and repair revealed from molecular epidemiology studies

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    Dusinska, Maria, E-mail: Maria.DUSINSKA@nilu.no [CEE-Health Effects Group, NILU - Norwegian Institute for Air Research, Kjeller (Norway); Staruchova, Marta; Horska, Alexandra [Department of Experimental and Applied Genetics, Slovak Medical University, Bratislava (Slovakia); Smolkova, Bozena [Laboratory of Cancer Genetics, Cancer Research Institute of the Slovak Academy of Sciences, Bratislava (Slovakia); Collins, Andrew [Department of Nutrition, Faculty of Medicine, University of Oslo (Norway); Bonassi, Stefano [Unit of Clinical and Molecular Epidemiology, IRCCS San Raffaele Pisana, Rome (Italy); Volkovova, Katarina [Department of Experimental and Applied Genetics, Slovak Medical University, Bratislava (Slovakia)

    2012-08-01

    Glutathione S-transferases (GSTs) are members of a multigene family of isoenzymes that are important in the control of oxidative stress and in phase II metabolism. Acting non-enzymically, GSTs can modulate signalling pathways of cell proliferation, cell differentiation and apoptosis. Using a molecular epidemiology approach, we have investigated a potential involvement of GSTs in DNA damage processing, specifically the modulation of DNA repair in a group of 388 healthy adult volunteers; 239 with at least 5 years of occupational exposure to asbestos, stone wool or glass fibre, and 149 reference subjects. We measured DNA damage in lymphocytes using the comet assay (alkaline single cell gel electrophoresis): strand breaks (SBs) and alkali-labile sites, oxidised pyrimidines with endonuclease III, and oxidised purines with formamidopyrimidine DNA glycosylase. We also measured GST activity in erythrocytes, and the capacity for base excision repair (BER) in a lymphocyte extract. Polymorphisms in genes encoding three GST isoenzymes were determined, namely deletion of GSTM1 and GSTT1 and single nucleotide polymorphism Ile105Val in GSTP1. Consumption of vegetables and wine correlated negatively with DNA damage and modulated BER. GST activity correlated with oxidised bases and with BER capacity, and differed depending on polymorphisms in GSTP1, GSTT1 and GSTM1. A significantly lower BER rate was associated with the homozygous GSTT1 deletion in all asbestos site subjects and in the corresponding reference group. Multifactorial analysis revealed effects of sex and exposure in GSTP1 Ile/Val heterozygotes but not in Ile/Ile homozygotes. These variants affected also SBs levels, mainly by interactions of GSTP1 genotype with exposure, with sex, and with smoking habit; and by an interaction between sex and smoking. Our results show that GST polymorphisms and GST activity can apparently influence DNA stability and repair of oxidised bases, suggesting a potential new role for these