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1.
Lett Appl Microbiol ; 64(4): 289-296, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28083911

RESUMEN

Methomyl (S-methyl N-(methylcarbamoyloxy) thioacetimidate) is a kind of oxime carbamate insecticide. It is considered to be extremely toxic to nontarget organism. To date, no pure culture or consortium has been reported to have the ability to degrade methomyl completely. In this study, a methomyl-degrading enrichment E1 was obtained by using the sludge from the wastewater-treating system of a pesticide manufacturer as the original inoculant. Two bacterial strains named MDW-2 and MDW-3 were isolated from this enrichment, and they were preliminarily identified as Aminobacter sp. and Afipia sp. respectively. Strains MDW-2 and MDW-3 could coexist and degrade 50 mg l-1 methomyl completely within 3 days by the cooperative metabolism. Methomyl was first converted to methomyl oxime and methylcarbamic acid by strain MDW-2, and the latter could be used as the carbon source for the growth of strain MDW-2. But methomyl oxime could not be sequentially degraded by strain MDW-2. However, it could be degraded and used as the carbon source by strain MDW-3. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents a bacterial combination of Aminobacter sp. MDW-2 and Afipia sp. MDW-3, which could degrade methomyl completely by biochemical cooperation. This study also proposes the biodegradation pathway of methomyl for the first time and highlights the application potential of a bacterial combination in the remediation of methomyl-contaminated environments.


Asunto(s)
Afipia/metabolismo , Insecticidas/metabolismo , Metomil/metabolismo , Phyllobacteriaceae/metabolismo , Afipia/genética , Biodegradación Ambiental , Carbamatos/química , Carbamatos/metabolismo , Insecticidas/química , Metomil/análogos & derivados , Metomil/química , Phyllobacteriaceae/genética , Filogenia , Análisis de Secuencia de ADN
2.
Cell Physiol Biochem ; 39(5): 1941-1954, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27771709

RESUMEN

BACKGROUND/AIMS: In previous publications we were able to demonstrate the exposure of phosphatidylserine (PS) in the outer membrane leaflet after activation of red blood cells (RBCs) by lysophosphatidic acid (LPA), phorbol-12 myristate-13acetate (PMA), or 4-bromo-A23187 (A23187). It has been concluded that three different mechanisms are responsible for the PS exposure in human RBCs: (i) Ca2+-stimulated scramblase activation (and flippase inhibition) by A23187, LPA, and PMA; (ii) PKCα activation by LPA and PMA; and (iii) enhanced lipid flip flop caused by LPA. Further studies aimed to elucidate interconnections between the increased Ca2+ content, scramblase- and PKCα-activation. In addition, the role of the Ca2+-activated K+ channel (Gardos channel) activity in the process of PS exposure needs to be investigated. METHODS: The intracellular Ca2+ content and the PS exposure of RBCs have been investigated after treatment with LPA (2.5 µM), PMA (6 µM), or A23187 (2 µM). Fluo-4 and annexin V-FITC has been used to detect intracellular Ca2+ content and PS exposure, respectively. Both parameters (Ca2+ content, PS exposure) were studied using flow cytometry. Inhibitors of the scramblase, the PKCα, and the Gardos channel have been applied. RESULTS: The percentage of RBCs showing PS exposure after activation with LPA, PMA, or A23187 is significantly reduced after inhibition of the scramblase using the specific inhibitor R5421 as well as after the inhibition of the PKCα using chelerythrine chloride or calphostin C. The inhibitory effect is more pronounced when the scramblase and the PKCα are inhibited simultaneously. Additionally, the inhibition of the Gardos channel using charybdotoxin resulted in a significant reduction of the percentage of RBCs showing PS exposure under all conditions measured. Similar results were obtained when the Gardos channel activity was suppressed by increased extracellular K+ content. CONCLUSION: PS exposure is mediated by the Ca2+-dependent scramblase but also by PKCα activated by LPA and PMA in a Ca2+-dependent and a Ca2+-independent manner. Furthermore, we hypothesize that a hyperpolarisation of RBCs caused by the opening of the Gardos channel is essential for the scramblase activity as well as for a fraction of the LPA-induced Ca2+ entry.


Asunto(s)
Calcimicina/farmacología , Calcio/metabolismo , Lisofosfolípidos/farmacología , Fosfatidilserinas/metabolismo , Acetato de Tetradecanoilforbol/farmacología , Anexina A5/genética , Anexina A5/metabolismo , Benzofenantridinas/farmacología , Células Cultivadas , Caribdotoxina/farmacología , Recuento de Eritrocitos , Eritrocitos , Regulación de la Expresión Génica , Humanos , Canales de Potasio de Conductancia Intermedia Activados por el Calcio/antagonistas & inhibidores , Canales de Potasio de Conductancia Intermedia Activados por el Calcio/genética , Canales de Potasio de Conductancia Intermedia Activados por el Calcio/metabolismo , Metomil/análogos & derivados , Metomil/farmacología , Naftalenos/farmacología , Fosfatidilserinas/química , Proteínas de Transferencia de Fosfolípidos/antagonistas & inhibidores , Proteínas de Transferencia de Fosfolípidos/genética , Proteínas de Transferencia de Fosfolípidos/metabolismo , Proteína Quinasa C-alfa/antagonistas & inhibidores , Proteína Quinasa C-alfa/genética , Proteína Quinasa C-alfa/metabolismo , Transducción de Señal
3.
Br J Pharmacol ; 177(17): 4007-4020, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-32496597

RESUMEN

BACKGROUND AND PURPOSE: Ethaninidothioic acid (R5421) has been used as a scramblase inhibitor to determine the role of phospholipid scrambling across a range of systems including platelet procoagulant activity. The selectivity of R5421 has not been thoroughly studied. Here, we characterised the effects of R5421 on platelet function and its suitability for use as a scramblase inhibitor. EXPERIMENTAL APPROACH: Human platelet activation was measured following pretreatment with R5421 and stimulation with a range of agonists. Phosphatidylserine exposure was measured using annexin V binding. Integrin αIIb ß3 activation and α-granule release were measured by flow cytometry. Cytosolic Ca2+ signals were measured using Cal520 fluorescence. An in silico ligand-based screen identified 16 compounds which were tested in these assays. KEY RESULTS: R5421 inhibited A23187-induced phosphatidylserine exposure in a time- and temperature-dependent manner. R5421 inhibited Ca2+ signalling from the PAR1, PAR4 and glycoprotein VI receptors as well as platelet αIIb ß3 integrin activation and α-granule release. R5421 is therefore not a selective inhibitor of platelet scramblase activity. An in silico screen identified the pesticide thiodicarb as similar to R5421. It also inhibited platelet phosphatidylserine exposure, Ca2+ signalling from the PAR1 and glycoprotein VI, αIIb ß3 activation and α-granule release. Thiodicarb additionally disrupted Ca2+ homeostasis in unstimulated platelets. CONCLUSION AND IMPLICATIONS: R5421 is not a selective inhibitor of platelet scramblase activity. We have identified the pesticide thiodicarb, which had similar effects on platelet function to R5421 as well as additional disruption of Ca2+ signalling which may underlie some of thiodicarb's toxicity.


Asunto(s)
Plaquetas , Proteínas de Transferencia de Fosfolípidos , Plaquetas/metabolismo , Humanos , Metomil/análogos & derivados , Fosfatidilserinas , Activación Plaquetaria , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo
4.
Environ Mol Mutagen ; 23(4): 306-11, 1994.
Artículo en Inglés | MEDLINE | ID: mdl-8013478

RESUMEN

The carbamate insecticide methomyl and the methomyl-containing technical formulation "Lannate 25" were tested on whole blood human lymphocyte cultures. Both products induced dose-dependent increases in chromosome aberrations and micronuclei. Lannate 25 induced DNA damage as measured by the alkaline elution assay and hydroxylation of guanine at the C8 position. Sister chromatid exchanges were not increased significantly with either product. Overall, the technical formulation was more active than the pure compound, when compared at similar concentrations of active principle. Moreover, a different ratio of CREST-positive/CREST-negative micronuclei was observed with the two products, pure methomyl being relatively more active than Lannate 25 in the induction of CREST-positive micronuclei. On the basis of these results, previous evaluations of methomyl as a nongenotoxic compound should be reconsidered.


Asunto(s)
Metomil/toxicidad , Mutagénesis , Mutágenos/toxicidad , 8-Hidroxi-2'-Desoxicoguanosina , Células Cultivadas , Centrómero , Distribución de Chi-Cuadrado , Aberraciones Cromosómicas , Daño del ADN , Desoxiguanosina/análogos & derivados , Desoxiguanosina/biosíntesis , Relación Dosis-Respuesta a Droga , Humanos , Modelos Lineales , Linfocitos/efectos de los fármacos , Metomil/análogos & derivados , Micronúcleos con Defecto Cromosómico/química , Pruebas de Micronúcleos , Proteínas Nucleares/análisis , Intercambio de Cromátides Hermanas
5.
Environ Mol Mutagen ; 24(3): 235-42, 1994.
Artículo en Inglés | MEDLINE | ID: mdl-7957126

RESUMEN

The carbamate insecticide, methomyl, and the methomyl-containing technical formulation, "Lannate 25", were tested for the induction of DNA damage in vivo. Swiss CD1 mice were treated intraperitoneally with test substances and the following tests were performed: alkaline elution of liver and kidney DNA, 8-hydroxyguanosine detection in liver DNA, and 32P-postlabelling analysis of DNA adducts in liver DNA. The clastogenic activity of the two pesticide preparations was also evaluated as micronucleus frequency in bone marrow. No DNA adducts were detected in liver DNA of mice treated with pure methomyl, while a dose-related increase in DNA adducts was found in Lannate 25-treated animals. All other tests were positive with both methomyl and Lannate 25. A summary of genotoxic activity of methomyl is also presented. The hypothesis that the observed genotoxic effects of methomyl are induced indirectly, through formation of active oxygen species, is discussed.


Asunto(s)
Daño del ADN , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Metomil/toxicidad , Mutagénesis/efectos de los fármacos , Animales , Médula Ósea/metabolismo , Células de la Médula Ósea , Núcleo Celular/efectos de los fármacos , Cromatografía en Capa Delgada , ADN/genética , ADN/metabolismo , Aductos de ADN/genética , Aductos de ADN/metabolismo , Daño del ADN/genética , Relación Dosis-Respuesta a Droga , Femenino , Guanosina/análogos & derivados , Guanosina/metabolismo , Inyecciones Intraperitoneales , Riñón/química , Riñón/citología , Hígado/química , Hígado/citología , Masculino , Metomil/administración & dosificación , Metomil/análogos & derivados , Ratones , Pruebas de Micronúcleos , Mutagénesis/genética , Control de Plagas , Radioisótopos de Fósforo , Especies Reactivas de Oxígeno
6.
Forensic Sci Int ; 227(1-3): 103-5, 2013 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-22999231

RESUMEN

We applied energy dispersive X-ray fluorescence spectrometry (EDX) in a case of poisoning by methomyl, a carbamate pesticide. Quantitative GC/MS analysis showed that the concentration of methomyl-oxime in the femoral blood was 4.0 µg/ml. The elemental analysis by EDX identified the high peak of silicon and sulfur in the stomach contents. We concluded that the cause of his death was methomyl poisoning. This indicates that screening of stomach contents by EDX provides useful information for the forensic diagnosis.


Asunto(s)
Insecticidas/envenenamiento , Metomil/envenenamiento , Espectrometría por Rayos X/métodos , Anciano de 80 o más Años , Toxicología Forense , Cromatografía de Gases y Espectrometría de Masas , Contenido Digestivo/química , Humanos , Insecticidas/análisis , Masculino , Metomil/análogos & derivados , Metomil/análisis , Metomil/sangre
8.
Arch Environ Contam Toxicol ; 10(1): 55-67, 1981.
Artículo en Inglés | MEDLINE | ID: mdl-7235742

RESUMEN

The penetration and fate of methomyl and its oxime metabolite, methomyl oxime or S-methyl N-hydroxythioacetimidate, were examined in house flies (Musca domestica Linnaeus), face flies (Musca autumnalis De Geer), black cutworm larvae (Agrotis ipsilon (Hufnagel), and twospotted spider mites (Tetranychus urticae Koch). Generally, penetration and metabolism of methomyl-1-14C were rapid in insects and spider mites although differences in rates among the different test organisms were observed. Metabolites from methomyl included CO2, methomyl oxime, and several unknowns. Penetration and metabolism of methomyl oxime-1-14C also were rapid in insects. Metabolites from methomyl oxime included a small amount of CO2 and several unidentified compounds. Organosoluble metabolites from methomyl oxime generally displayed similar chromatographic behavior as those from methomyl. Methomyl was hydrolyzed to its oxime metabolite, which, apparently, was further metabolized to 14CO2 presumably via a Beckmann rearrangement reaction.


Asunto(s)
Insectos/metabolismo , Insecticidas/metabolismo , Metomil/metabolismo , Ácaros/metabolismo , Animales , Femenino , Cinética , Masculino , Metomil/análogos & derivados
9.
Blood ; 91(6): 2133-8, 1998 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-9490700

RESUMEN

Scott syndrome is an hereditary bleeding disorder characterized by a deficiency in platelet procoagulant activity. Unlike normal blood cells, Scott platelets, as well as erythrocytes and lymphocytes, are strongly impaired in their ability to scramble their membrane phospholipids when challenged with Ca2+. In normal cells this collapse of membrane asymmetry leads to surface exposure of phosphatidylserine. Here we report that Scott erythrocytes show an apparent defect in tyrosine phosphorylation on treatment with Ca2+-ionophore. Diminished tyrosine phosphorylation was also apparent in activated Scott platelets, but much less pronounced than observed in red blood cells. On the other hand, tyrosine phosphorylation profiles observed in Scott red blood cell ghosts after sealing in the presence of adenosine triphosphate (ATP) were indistinguishable from those obtained from normal ghosts. Several observations argue in favor of a mechanism in which tyrosine phosphorylation in red blood cells is facilitated by, rather than required for scrambling of membrane lipids. Staurosporin blocks tyrosine phosphorylation in normal red blood cells, but does not inhibit the lipid scrambling process. White ghosts from normal erythrocytes, resealed in the absence of ATP, exhibit Ca2+-induced lipid scrambling without tyrosine phosphorylation. A selective inhibitor of Ca2+-induced lipid scrambling also showed an apparent inhibition of tyrosine phosphorylation in ionophore-treated normal red blood cells, similar to that observed in Scott erythrocytes. While this inhibitor also suppressed Ca2+-induced lipid scrambling in ghosts that were sealed in the presence of ATP, it did not inhibit tyrosine kinase activity. We conclude that the apparent deficiency in tyrosine phosphorylation in Scott cells is an epiphenomenon, possibly associated with a defect in phospholipid scrambling, but not causal to this defect.


Asunto(s)
Trastornos de la Coagulación Sanguínea/genética , Calcio/farmacología , Proteínas Portadoras/antagonistas & inhibidores , Membrana Eritrocítica/efectos de los fármacos , Ionóforos/farmacología , Lípidos de la Membrana/metabolismo , Proteínas de la Membrana/antagonistas & inhibidores , Metomil/análogos & derivados , Proteínas de Transferencia de Fosfolípidos , Fosfolípidos/metabolismo , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Proteínas Tirosina Quinasas/sangre , Adenosina Trifosfato/farmacología , Trastornos de la Coagulación Sanguínea/sangre , Plaquetas/efectos de los fármacos , Plaquetas/metabolismo , Proteínas Portadoras/sangre , Proteínas Portadoras/genética , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Inhibidores Enzimáticos/farmacología , Membrana Eritrocítica/metabolismo , Humanos , Proteínas de la Membrana/sangre , Proteínas de la Membrana/genética , Metomil/farmacología , Fosfatidilserinas/sangre , Fosfatidilserinas/fisiología , Fosforilación/efectos de los fármacos , Estaurosporina/farmacología , Síndrome , Tromboplastina/metabolismo
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