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  1. Direct Analysis of Glyphosate, Glufosinate, and Their Metabolites in Palm Oil Using Liquid Chromatography with Tandem Mass Spectrometry
    Shinde R, Halim N, Banerjee AK
    J AOAC Int, 2020 Nov 01;103(6):1528-1533.
    PMID: 33247748 DOI: 10.1093/jaoacint/qsaa066
    BACKGROUND: Glyphosate and glufosinate are broad-spectrum herbicides which are frequently used in palm oil plantations for weed control. Metabolites of these herbicides are known to have environmental and food safety implications. As there is no validated method for multiresidue testing of these herbicides and their metabolites in palm oil products, a new method was needed for the purpose of regulatory analysis.

    OBJECTIVE: In this study, we endeavored to develop a rapid method for multiresidue analysis of glyphosate (+aminomethylphosphonic acid) and glufosinate (+3-methylphosphinicopropionic acid and N-acetyl-glufosinate) in refined and crude palm oil matrices using liquid chromatography (LC) tandem mass spectrometry (MS/MS).

    METHOD: The optimized sample preparation workflow included extraction of refined or crude palm oil (10 g) with acidified water (0.1 M HCl), cleanup by phase separation with dichloromethane, and analysis by LC-MS/MS with multiple reaction monitoring.

    RESULTS: The use of a Torus-DEA LC column ensured simultaneous analysis of these compounds within a runtime of 10 min. The LOQ of these analytes was 0.01 mg/kg, except that of aminomethylphosphonic acid which was 0.02 mg/kg. The method sensitivity complied with the national maximum residue limits of Malaysia and the European Union. Also, the method selectivity, sensitivity, accuracy, and precision were aligned with the SANTE/12682/2019 guidelines of analytical quality control.

    CONCLUSIONS: The potentiality of the optimized method lies in a high throughput direct analysis of glyphosate and glufosinate with their metabolites in a single chromatographic run. The method is fit for purpose for regulatory testing of these residues in a broad range of palm oil matrices.

    HIGHLIGHTS: The study reports for the first time a validated method for simultaneous analysis of glyphosate, glufosinate, and their metabolites in a range of palm oil products. The method did not require a derivatization step and provided a high throughput analysis of these compounds with satisfactory selectivity, sensitivity, accuracy, and precision.

    Matched MeSH terms: Glycine/analogs & derivatives
  2. Modeling biodegradation and kinetics of glyphosate by artificial neural network
    Nourouzi MM, Chuah TG, Choong TS, Rabiei F
    J Environ Sci Health B, 2012;47(5):455-65.
    PMID: 22424071 DOI: 10.1080/03601234.2012.663603
    An artificial neural network (ANN) model was developed to simulate the biodegradation of herbicide glyphosate [2-(Phosphonomethylamino) acetic acid] in a solution with varying parameters pH, inoculum size and initial glyphosate concentration. The predictive ability of ANN model was also compared with Monod model. The result showed that ANN model was able to accurately predict the experimental results. A low ratio of self-inhibition and half saturation constants of Haldane equations (< 8) exhibited the inhibitory effect of glyphosate on bacteria growth. The value of K(i)/K(s) increased when the mixed inoculum size was increased from 10(4) to 10(6) bacteria/mL. It was found that the percentage of glyphosate degradation reached a maximum value of 99% at an optimum pH 6-7 while for pH values higher than 9 or lower than 4, no degradation was observed.
    Matched MeSH terms: Glycine/analogs & derivatives*
  3. Genetic diversity and in silico evidence of target-site mutation in the EPSPS gene in endowing glyphosate resistance in Eleusine indica (L.) from Malaysia
    Franci J, Lam KW, Chuah TS, Cha TS
    Pestic Biochem Physiol, 2020 May;165:104556.
    PMID: 32359543 DOI: 10.1016/j.pestbp.2020.104556
    Glyphosate-resistant populations of Eleusine indica are widespread in several states of Malaysia. A whole-plant bioassay confirmed that eight out of the 17 populations tested were resistant to glyphosate at double the recommended rate of 2.44 kg ha-1. Screening with allele-specific PCR (AS-PCR) revealed that resistant plants contained an EPSPS gene with either the homozygous S/S-106 or the heterozygous P/S-106 alleles. All susceptible plants contained only the homozygous P/P-106 allele. In addition, DNA sequences of the full-length EPSPS gene from one susceptible (SB) and four resistant (R2, R6, R8 and R11) populations revealed an amino acid substitution of T102I in all the resistant plants, while another substitution of P381L was only found in resistant populations R6 and R11. The significance of the P381L mutation was examined by Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA) and residue interaction network (RIN) analyses, which suggests the P381L mutation may contribute to resistance. Mutations at 102 and 106 occur widely in the EPSPS gene of glyphosate-resistant E. indica populations from Malaysia with the TIPS mutation. In addition, the P381L mutation could also contribute to resistance.
    Matched MeSH terms: Glycine/analogs & derivatives
  4. Effect of glyphosate-based herbicide on early life stages of Java medaka (Oryzias javanicus): a potential tropical test fish
    Yusof S, Ismail A, Alias MS
    Mar Pollut Bull, 2014 Aug 30;85(2):494-8.
    PMID: 24731878 DOI: 10.1016/j.marpolbul.2014.03.022
    Glyphosate is globally a widely used herbicide, yet there is little information on their toxicity to marine fishes. Java medaka, a small tropical fish native to coastal areas in several Southeast Asian countries, is viewed as a suitable candidate for toxicity test and thus was used for this study. Java medaka adults were cultured in the laboratory and the fertilized eggs of the F2 generation were exposed to different concentrations of glyphosate-based herbicide (100, 200, 300, 400 and 500 ppm) until they hatched. The survival and hatching rates of the embryos, changes in the heart rate and morphological impairments were recorded. Generally, survival and hatching percentage decreased as glyphosate concentration increased. Absence of pectoral fin(s) and cornea, permanently bent tail, irregular shaped abdomen, and cell disruption in the fin, head and abdomen are among the common teratogenic effects observed. Furthermore, risk factor also increased with the increased in glyphosate concentrations.
    Matched MeSH terms: Glycine/analogs & derivatives*
  5. Identification and characterization of RAPD-SCAR markers linked to glyphosate-susceptible and -resistant biotypes of Eleusine indica (L.) Gaertn
    Cha TS, Anne-Marie K, Chuah TS
    Mol Biol Rep, 2014 Feb;41(2):823-31.
    PMID: 24374894 DOI: 10.1007/s11033-013-2922-7
    Eleusine indica is one of the most common weed species found in agricultural land worldwide. Although herbicide-glyphosate provides good control of the weed, its frequent uses has led to abundant reported cases of resistance. Hence, the development of genetic markers for quick detection of glyphosate-resistance in E. indica population is imperative for the control and management of the weed. In this study, a total of 14 specific random amplified polymorphic DNA (RAPD) markers were identified and two of the markers, namely S4R727 and S26R6976 were further sequence characterized. Sequence alignment revealed that marker S4R727 showing a 12-bp nucleotides deletion in resistant biotypes, while marker S26R6976 contained a 167-bp nucleotides insertion in the resistant biotypes. Based on these sequence differences, three pairs of new sequence characterized amplified region (SCAR) primers were developed. The specificity of these primer pairs were further validated with genomic DNA extracted from ten individual plants of one glyphosate-susceptible and five glyphosate-resistant (R2, R4, R6, R8 and R11) populations. The resulting RAPD-SCAR markers provided the basis for assessing genetic diversity between glyphosate-susceptible and -resistant E. indica biotypes, as well for the identification of genetic locus link to glyphosate-resistance event in the species.
    Matched MeSH terms: Glycine/analogs & derivatives
  6. Rapid and direct determination of glyphosate, glufosinate, and aminophosphonic acid by online preconcentration CE with contactless conductivity detection
    See HH, Hauser PC, Ibrahim WA, Sanagi MM
    Electrophoresis, 2010 Jan;31(3):575-82.
    PMID: 20119968 DOI: 10.1002/elps.200900380
    Rapid and direct online preconcentration followed by CE with capacitively coupled contactless conductivity detection (CE-C(4)D) is evaluated as a new approach for the determination of glyphosate, glufosinate (GLUF), and aminophosphonic acid (AMPA) in drinking water. Two online preconcentration techniques, namely large volume sample stacking without polarity switching and field-enhanced sample injection, coupled with CE-C(4)D were successfully developed and optimized. Under optimized conditions, LODs in the range of 0.01-0.1 microM (1.7-11.1 microg/L) and sensitivity enhancements of 48- to 53-fold were achieved with the large volume sample stacking-CE-C(4)D method. By performing the field-enhanced sample injection-CE-C(4)D procedure, excellent LODs down to 0.0005-0.02 microM (0.1-2.2 microg/L) as well as sensitivity enhancements of up to 245- to 1002-fold were obtained. Both techniques showed satisfactory reproducibility with RSDs of peak height of better than 10%. The newly established approaches were successfully applied to the analysis of glyphosate, glufosinate, and aminophosphonic acid in spiked tap drinking water.
    Matched MeSH terms: Glycine/analogs & derivatives*
  7. FTIR, CHNS and XRD analyses define mechanism of glyphosate herbicide removal by electrocoagulation
    Danial R, Sobri S, Abdullah LC, Mobarekeh MN
    Chemosphere, 2019 Oct;233:559-569.
    PMID: 31195261 DOI: 10.1016/j.chemosphere.2019.06.010
    In this study, the performance of glyphosate removal in an electrocoagulation batch with two electrodes formed by the same metal type, consisting of aluminum, iron, steel and copper have been compared. The aim of this study intends to remove glyphosate from an aqueous solution by an electrocoagulation process using metal electrode plates, which involves electrogeneration of metal cations as coagulant agents. The production of metal cations showed an ability to bind together to form aggregates of flocs composed of a combination of glyphosate and metal oxide. Electrocoagulation using aluminum electrodes indicated a high percentage removal of glyphosate, 94.25%; followed by iron electrodes, 88.37%; steel electrodes, 62.82%; and copper electrodes, 46.69%. The treated aqueous solution was then analyzed by Fourier Transform Infrared Spectroscopy. Percentages of Carbon, Hydrogen, Nitrogen, Sulfur remaining in the treated aqueous solution after the electrocoagulation process have been determined. The treated water and sludge were characterized and the mechanism of the overall process was concluded as an outcome. An X-Ray Diffraction analysis of dried sludge confirmed that new polymeric compounds were formed during the treatment. The sludge composed of new compounds were also verified the removals. This study revealed that an electrocoagulation process using metal electrodes is reliable and efficient.
    Matched MeSH terms: Glycine/analogs & derivatives*
  8. Does maternal-fetal transfer of creatine occur in pregnant sheep?
    Baharom S, De Matteo R, Ellery S, Della Gatta P, Bruce CR, Kowalski GM, et al.
    Am J Physiol Endocrinol Metab, 2017 07 01;313(1):E75-E83.
    PMID: 28325734 DOI: 10.1152/ajpendo.00450.2016
    Our aim was to determine the disposition of creatine in ovine pregnancy and whether creatine is transferred across the placenta from mother to fetus. Pregnant ewes received either1) a continuous intravenous infusion of creatine monohydrate or saline from 122 to 131 days gestation, with maternal and fetal arterial blood and amniotic fluid samples collected daily for creatine analysis and fetal tissues collected at necropsy at 133 days for analysis of creatine content, or2) a single systemic bolus injection of [13C]creatine monohydrate at 130 days of gestation, with maternal and fetal arterial blood, uterine vein blood, and amniotic fluid samples collected before and for 4 h after injection and analyzed for creatine, creatine isotopic enrichment, and guanidinoacetic acid (GAA; precursor of creatine) concentrations. Presence of the creatine transporter-1 (SLC6A8) and l-arginine:glycine amidinotransferase (AGAT; the enzyme synthesizing GAA) proteins were determined by Western blots of placental cotyledons. The 10-day creatine infusion increased maternal plasma creatine concentration three- to fourfold (P< 0.05) without significantly changing fetal arterial, amniotic fluid, fetal tissues, or placental creatine content. Maternal arterial13C enrichment was increased (P< 0.05) after bolus [13C]creatine injection without change of fetal arterial13C enrichment. SLC6A8 and AGAT proteins were identified in placental cotyledons, and GAA concentration was significantly higher in uterine vein than maternal artery plasma. Despite the presence of SLC6A8 protein in cotyledons, these results suggest that creatine is not transferred from mother to fetus in near-term sheep and that the ovine utero-placental unit releases GAA into the maternal circulation.
    Matched MeSH terms: Glycine/analogs & derivatives*
  9. Adsorption-desorption and leaching potential of glyphosate and aminomethylphosphonic acid in acidic Malaysian soil amended with cow dung and rice husk ash
    Garba J, Samsuri AW, Othman R, Ahmad Hamdani MS
    Environ Monit Assess, 2018 Oct 27;190(11):676.
    PMID: 30368595 DOI: 10.1007/s10661-018-7034-3
    This study investigates adsorption-desorption and the leaching potential of glyphosate and aminomethylphosphonic acid (AMPA) in control and amended-addition of cow dung or rice husk ash-acidic Malaysian soil with high oxide mineral content. The addition of cow dung or rice husk ash increased the adsorptive removal of AMPA. The isotherm data of glyphosate and AMPA best fitted the Freundlich model. The constant Kf for glyphosate was high in the control soil (544.873 mg g-1) followed by soil with cow dung (482.451 mg g-1) then soil with rice husk ash (418.539 mg g-1). However, for AMPA, soil with cow dung was high (166.636 mg g-1) followed by soil with rice husk ash (137.570 mg g-1) then the control soil (48.446 mg g-1). The 1/n values for both glyphosate and AMPA adsorptions were
    Matched MeSH terms: Glycine/analogs & derivatives*
  10. The oral and intratracheal toxicities of ROUNDUP and its components to rats
    Adam A, Marzuki A, Abdul Rahman H, Abdul Aziz M
    Vet Hum Toxicol, 1997 Jun;39(3):147-51.
    PMID: 9167243
    The toxicities of ROUNDUP and its component chemicals, glyphosate (N-phosphonomethylglycine) and polyoxyethyleneamine (POEA), were determined at 0, 1, 3, 6 and 24 h following administration to rats. The intratracheal administration of glyphosate (0.2 g/kg), POEA (0.1 g/kg), a mixture of glyphosate (0.2 g/kg) + POEA (0.1 g/kg), or ROUNDUP (containing 0.2 g/kg glyphosate and 0.1 g/kg POEA) elicited immediate respiratory effects which were more severe and which lasted longer in the groups receiving the POEA-containing preparations than in the glyphosate alone group. By 1 h, all test preparations had caused deaths, but more occurred from the POEA-containing preparations than from glyphosate. The po administration of POEA (1 g/kg), the mixture of glyphosate (2 g/kg) +POEA (1 g/kg), or ROUNDUP (containing 2 g/kg glyphosate and 1 g/kg POEA) produced diarrhea and blood-stained weeping from noses. Death was only seen from POEA at 24 h. Glyphosate (2 g/kg po) produced transient diarrhea without nose bleeds; POEA caused diarrhea at 1 h; and the mixture of POEA + glyphosate produced diarrhea later that increased in severity with time. Bloody nose secretions were seen only with the preparations that contained POEA. No deaths, respiratory effects or bloody nose secretions occurred in controls given saline. Both POEA and glyphosate caused lung hemorrhages and lung epithelial cell damage with po or intratracheal exposures. These results indicate POEA and preparations that contained POEA were more toxic than glyphosate.
    Matched MeSH terms: Glycine/analogs & derivatives*
  11. Green nanoemulsion-laden glyphosate isopropylamine formulation in suppressing creeping foxglove (A. gangetica), slender button weed (D. ocimifolia) and buffalo grass (P. conjugatum)
    Lim CJ, Basri M, Omar D, Abdul Rahman MB, Salleh AB, Raja Abdul Rahman RN
    Pest Manag Sci, 2013 Jan;69(1):104-11.
    PMID: 22865686 DOI: 10.1002/ps.3371
    Pesticides are developed with carriers to improve their physicochemical properties and, accordingly, the bioefficacy of the applied formulation. For foliar-applied herbicide, generally less than 0.1% of the active ingredient reaching the target site could reduce pesticide performance. Recently, a carrier of nanoemulsion consisting of oil, surfactant and water, with a particle size of less than 200 nm, has been shown to enhance drug permeability for skin penetration in pharmaceutical delivery systems. In the present work, the aim was to formulate a water-soluble herbicide, glyphosate isopropylamine (IPA), using a green nanoemulsion system for a biological activity study against the weeds creeping foxglove, slender button weed and buffalo grass.
    Matched MeSH terms: Glycine/analogs & derivatives*
  12. Characterization of Eleusine indica with gene mutation or amplification in EPSPS to glyphosate
    Chen J, Jiang C, Huang H, Wei S, Huang Z, Wang H, et al.
    Pestic Biochem Physiol, 2017 Nov;143:201-206.
    PMID: 29183593 DOI: 10.1016/j.pestbp.2017.09.012
    The evolution of weed-resistant species threatens the sustainable use of glyphosate, which is the most important herbicide widely used in agriculture worldwide. Moreover, the high glyphosate resistance (>180-fold based on LD50) of Eleusine indica found in Malaysia, which carries a double mutation in its 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), made the control of this species more difficult. By contrast, the same species carrying the same double mutation in EPSPS (T102I+P106S) but found in China only shows a resistance level of not more than 14-fold based on GR50. The resistance level of this population is four times higher than that of the population carrying a single mutation (P106L). Although the members of this population survive under a high glyphosate dosage of 10,080gaeha-1, their growth was significantly inhibited by glyphosate under the recommend dose (840gaeha-1), where in the fresh weight was 85.4% of the control. EPSPS expression, relative copy number, and EPSPS activity in this population were similar to those of the susceptible population. In addition, the expression of two glutathione transferase (GST) genes (GST-U8 and GST-23) and the enzyme activity of the GST in this population did not significantly differ from those of the susceptible population. This finding is important in elucidating the resistance of the naturally evolved glyphosate-resistant (GR) weed species carrying a double mutation in EPSPS to glyphosate.
    Matched MeSH terms: Glycine/analogs & derivatives
  13. Biological and cytoselective anticancer properties of copper(II)-polypyridyl complexes modulated by auxiliary methylated glycine ligand
    Seng HL, Wang WS, Kong SM, Alan Ong HK, Win YF, Raja Abd Rahman RN, et al.
    Biometals, 2012 Oct;25(5):1061-81.
    PMID: 22836829 DOI: 10.1007/s10534-012-9572-4
    A series of ternary copper(II)-1,10-phenanthroline complexes with glycine and methylated glycine derivatives, [Cu(phen)(aa)(H(2)O)]NO(3)·xH(2)O 1-4 (amino acid (aa): glycine (gly), 1; DL: -alanine (DL: -ala), 2; 2,2-dimethylglycine (C-dmg), 3; sarcosine (sar), 4), were synthesized and characterized by FTIR, elemental analysis, electrospray ionization-mass spectra (ESI-MS), UV-visible spectroscopy and molar conductivity measurement. The determined X-ray crystallographic structures of 2 and 3 show each to consist of distorted square pyramidal [Cu(phen)(aa)(H(2)O)](+) cation, a nitrate counter anion, and with or without lattice water, similar to previously reported structure of [Cu(phen)(gly)(H(2)O)]NO(3)·1½H(2)O. It is found that 1-4 exist as 1:1 electrolytes in aqueous solution, and the cationic copper(II) complexes are at least stable up to 24 h. Positive-ion ESI-MS spectra show existence of only undissociated [Cu(phen)(aa)](+) species. Electron paramagnetic resonance, gel electrophoresis, fluorescence quenching, and restriction enzyme inhibition assay were used to study the binding interaction, binding affinity and selectivity of these complexes for various types of B-form DNA duplexes and G-quadruplex. All complexes can bind selectively to DNA by intercalation and electrostatic forces, and inhibit topoisomerase I. The effect of the methyl substituents of the coordinated amino acid in the above complexes on these biological properties are presented and discussed. The IC(50) values (24 h) of 1-4 for nasopharyngeal cancer cell line HK1 are in the range 2.2-5.2 μM while the corresponding values for normal cell line NP69 are greater than 13.0 μM. All complexes, at 5 μM, induced 41-60 % apoptotic cell death in HK1 cells but no significant cell death in NP69 cells.
    Matched MeSH terms: Glycine/analogs & derivatives
  14. Dynamic supported liquid membrane tip extraction of glyphosate and aminomethylphosphonic acid followed by capillary electrophoresis with contactless conductivity detection
    See HH, Hauser PC, Sanagi MM, Ibrahim WA
    J Chromatogr A, 2010 Sep 10;1217(37):5832-8.
    PMID: 20696433 DOI: 10.1016/j.chroma.2010.07.054
    A dynamic supported liquid membrane tip extraction (SLMTE) procedure for the effective extraction and preconcentration of glyphosate (GLYP) and its metabolite aminomethylphosphonic acid (AMPA) in water has been investigated. The SLMTE procedure was performed in a semi-automated dynamic mode and demonstrated a greater performance against a static extraction. Several important extraction parameters such as donor phase pH, cationic carrier concentration, type of membrane solvent, type of acceptor stripping phase, agitation and extraction time were comprehensively optimized. A solution of Aliquat-336, a cationic carrier, in dihexyl ether was selected as the supported liquid incorporated into the membrane phase. Quantification of GLYP and AMPA was carried out using capillary electrophoresis with contactless conductivity detection. An electrolyte solution consisting of 12 mM histidine (His), 8 mM 2-(N-morpholino)ethanesulfonic acid (MES), 75 microM cetyltrimethylammonium bromide (CTAB), 3% methanol, pH 6.3, was used as running buffer. Under the optimum extraction conditions, the method showed good linearity in the range of 0.01-200 microg/L (GLYP) and 0.1-400 microg/L (AMPA), acceptable reproducibility (RSD 5-7%, n=5), low limits of detection of 0.005 microg/L for GLYP and 0.06 microg/L for AMPA, and satisfactory relative recoveries (90-94%). Due to the low cost, the SLMTE device was disposed after each run which additionally eliminated the possibility of carry-over between runs. The validated method was tested for the analysis of both analytes in spiked tap water and river water with good success.
    Matched MeSH terms: Glycine/analogs & derivatives*
  15. Preconditioning effect of (S)-3,5-dihydroxyphenylglycine on ischemic injury in middle cerebral artery occluded Sprague-Dawley rats
    Nik Ramli NN, Omar N, Husin A, Ismail Z, Siran R
    Neurosci Lett, 2015 Feb 19;588:137-41.
    PMID: 25562631 DOI: 10.1016/j.neulet.2014.12.062
    Glutamate receptors are the integral cellular components associated with excitotoxicity mechanism induced by the ischemic cascade events. Therefore the glutamate receptors have become the major molecular targets of neuroprotective agents in stroke researches. Recent studies have demonstrated that a Group I metabotropic glutamate receptor agonist, (S)-3,5-dihydroxyphenylglycine ((S)-3,5-DHPG) preconditioning elicits neuroprotection in the hippocampal slice cultures exposed to toxic level of N-methyl-d-aspartate (NMDA). We further investigated the preconditioning effects of (S)-3,5-DHPG on acute ischemic stroke rats. One 10 or 100μM of (S)-3,5-DHPG was administered intrathecally to Sprague-Dawley adult male rats, 2h prior to induction of acute ischemic stroke by middle cerebral artery occlusion (MCAO). After 24h, neurological deficits were evaluated by modified stroke severity scores and grid-walking test. All rats were sacrificed and infarct volumes were determined by 2,3,5-triphenyltetrazolium chloride staining. The serum level of neuron-specific enolase (NSE) of each rat was analyzed by enzyme-linked immunosorbent assay (ELISA). One and 10μM of (S)-3,5-DHPG preconditioning in the stroke rats showed significant improvements in motor impairment (P<0.01), reduction in the infarct volume (P<0.01) and reduction in the NSE serum level (P<0.01) compared to the control stroke rats. We conclude that 1 and 10μM (S)-3,5-DHPG preconditioning induced protective effects against acute ischemic insult in vivo.
    Matched MeSH terms: Glycine/analogs & derivatives*
  16. LPS-induced Apoptosis is Partially Mediated by Hydrogen Sulphide in RAW 264.7 Murine Macrophages
    George L, Ramasamy T, Sirajudeen K, Manickam V
    Immunol Invest, 2019 Jul;48(5):451-465.
    PMID: 30689461 DOI: 10.1080/08820139.2019.1566355
    Lipopolysaccharide (LPS) induces apoptosis in murine macrophages through the autocrine secretion of tumor necrosis factor (TNF)-α and nitric oxide (NO). LPS-induced inflammation in murine macrophages is associated with hydrogen sulfide (H2S) production. In this present study, we reported the novel role of H2S in LPS-induced apoptosis and its underlying molecular mechanism specifically at late phases in murine macrophage cells. Stimulation of RAW 264.7 macrophages with LPS resulted in a time- and dose-dependent induction of apoptosis. We observed that the LPS-induced early apoptosis (associated with TNF-α secretion) in macrophages was not inhibited in the presence of H2S inhibitor (DL-propargylglycine), whereas early apoptosis was absent in the presence of TNF receptor antibody. Interestingly, LPS-induced late apoptosis paralleled with H2S production was reduced in the presence of H2S inhibitor but not with TNF receptor antibody. The late apoptotic events mediated by H2S and not the TNF-α induced early apoptosis correlated significantly with the induction of p53 and Bax expression in LPS-induced macrophages. Thus, it is possible that RAW 264.7 murine macrophages treated with LPS mediated early apoptosis through TNF-α and the late apoptotic events through the production of H2S.
    Matched MeSH terms: Glycine/analogs & derivatives
  17. Glyphosate-resistant goosegrass. Identification of a mutation in the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase
    Baerson SR, Rodriguez DJ, Tran M, Feng Y, Biest NA, Dill GM
    Plant Physiol, 2002 Jul;129(3):1265-75.
    PMID: 12114580
    The spontaneous occurrence of resistance to the herbicide glyphosate in weed species has been an extremely infrequent event, despite over 20 years of extensive use. Recently, a glyphosate-resistant biotype of goosegrass (Eleusine indica) was identified in Malaysia exhibiting an LD(50) value approximately 2- to 4-fold greater than the sensitive biotype collected from the same region. A comparison of the inhibition of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity by glyphosate in extracts prepared from the resistant (R) and sensitive (S) biotypes revealed an approximately 5-fold higher IC(50)(glyphosate) for the (R) biotype. Sequence comparisons of the predicted EPSPS mature protein coding regions from both biotypes revealed four single-nucleotide differences, two of which result in amino acid changes. One of these changes, a proline to serine substitution at position 106 in the (R) biotype, corresponds to a substitution previously identified in a glyphosate-insensitive EPSPS enzyme from Salmonella typhimurium. Kinetic data generated for the recombinant enzymes suggests that the second substitution identified in the (R) EPSPS does not contribute significantly to its reduced glyphosate sensitivity. Escherichia coli aroA- (EPSPS deficient) strains expressing the mature EPSPS enzyme from the (R) biotype exhibited an approximately 3-fold increase in glyphosate tolerance relative to strains expressing the mature EPSPS from the (S) biotype. These results provide the first evidence for an altered EPSPS enzyme as an underlying component of evolved glyphosate resistance in any plant species.
    Matched MeSH terms: Glycine/analogs & derivatives*
  18. Fulltext BACE1 inhibitory activity of fungal endophytic extracts from Malaysian medicinal plants
    Harun A, James RM, Lim SM, Abdul Majeed AB, Cole AL, Ramasamy K
    BMC Complement Altern Med, 2011 Sep 24;11:79.
    PMID: 21943123 DOI: 10.1186/1472-6882-11-79
    BACKGROUND: BACE1 was found to be the major β-secretase in neurons and its appearance and activity were found to be elevated in the brains of AD patients. Fungal endophytic extracts for BACE1 inhibitory activity and cytotoxicity against PC-12 (a rat pheochromocytoma with neuronal properties) and WRL68 (a non-tumorigenic human hepatic) were investigated.

    METHODS: Endophytes were isolated from plants collected from Kuala Pilah, Negeri Sembilan and the National Park, Pahang and the extracts were tested for BACE1 inhibition. For investigation of biological activity, the pure endophytic cultures were cultivated for 14 days on PDA plates at 28°C and underwent semipolar extraction with ethyl acetate.

    RESULTS: Of 212 endophytic extracts (1000 μg/ml), 29 exhibited more than 90% inhibition of BACE1 in the preliminary screening. Four extracts from isolates HAB16R13, HAB16R14, HAB16R18 and HAB8R24 identified as Cytospora rhizophorae were the most active with IC(50(BACE1)) values of less than 3.0 μg/ml. The most active extract HAB16R13 was shown to non-competitively inhibit BACE1 with K(i) value of 10.0 μg/ml. HAB16R13 was considered non-potent against PC-12 and WRL68 (IC(50(CT))) of 60.0 and 40.0 μg/ml, respectively).

    CONCLUSIONS: This first report on endophytic fungal extract with good BACE1 inhibitory activity demonstrates that more extensive study is required to uncover the potential of endophytes.

    Matched MeSH terms: Glycine/analogs & derivatives
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