RESUMEN
With the aim of explaining the variations in microcystin (MC) concentrations during cyanobacterial blooms, we studied several Microcystis aeruginosa populations blooming in different freshwater ecosystems located in the same geographical area. As assessed by real-time PCR, it appeared that the potentially MC-producing cells (mcyB(+)) were predominant (70 to 100%) in all of these M. aeruginosa populations, with the exception of one population in which non-MC-producing cells always dominated. Apart from the population in the Grangent Reservoir, we found that the proportions of potentially MC-producing and non-MC-producing cells varied little over time, which was consistent with the fact that according to a previous study of the same populations, the intergenic transcribed spacer (ITS) genotype composition did not change (38). In the Grangent Reservoir, the MC-RR variant was the dominant microcystin variant throughout the bloom season, despite changes in the ITS composition and in the proportions of mcyB(+) cells. Finally, the variations in total MC concentrations (0.3 to 15 microg liter(-1)) and in the MC cellular quotas (0.01 to 3.4 pg cell(-1)) were high both between and within sites, and no correlation was found between the MC concentrations and the proportion of mcyB(+) cells. All of these findings demonstrate that very different results can be found for the proportions of potentially MC-producing and non-MC-producing cells and MC concentrations, even in M. aeruginosa populations living in more or less connected ecosystems, demonstrating the importance of the effect of very local environmental conditions on these parameters and also the difficulty of predicting the potential toxicity of Microcystis blooms.
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Agua Dulce/microbiología , Microcistinas/análisis , Microcystis/química , Proteínas Bacterianas/genética , ADN Espaciador Ribosómico/genética , Microcystis/crecimiento & desarrollo , Reacción en Cadena de la PolimerasaRESUMEN
Highly viscous extracellular proteoglycan (EPG) has been isolated from culture medium of the unicellular red alga Rhodella grisea (Rhodophyceae) by ethanol precipitation. EPG was composed of xylose (29.3%), 3-O-methyl-xylose (26.0%), uronic acids (17.1%), rhamnose (14.4%), galactose (7.5%), glucose (3.9%), arabinose (1.4%) and mannose (0.4%), and traces of fucose, 4-O-methyl-xylose and 2,3-di-O-methyl-rhamnose or fucose. In addition, the polymer contained proteins (13.1%), sulphates and 13C-CP MAS spectra indicated the presence of acetyl and succinyl groups. The molecular mass was estimated to be 136,000. Ion-exchange chromatography afforded five fractions differing in composition of neutral sugars, uronic acids, and protein content indicating thus the complex structure of the EPG.
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Espacio Extracelular/metabolismo , Proteoglicanos/biosíntesis , Rhodophyta/metabolismo , Espectroscopía de Resonancia Magnética , Proteoglicanos/química , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
AIMS: The primary aim of this study was to analyse the position of the acetabular and femoral components in total hip arthroplasty undertaken using an anterior surgical approach. PATIENTS AND METHODS: In a prospective, single centre study, we used the EOS imaging system to analyse the position of components following THA performed via the anterior approach in 102 patients (103 hips) with a mean age of 64.7 years (sd 12.6). Images were taken with patients in the standing position, allowing measurement of both anatomical and functional anteversion of the acetabular component. RESULTS: The mean inclination of the acetabular component was 39° (standard deviation (sd) 6), the mean anatomical anteversion was 30° (sd 10), and the mean functional anteversion was 31° (sd 8) five days after surgery. The mean anteversion of the femoral component was 20° (sd 11). Anatomical and functional anteversion of the acetabular component differed by > 10° in 23 (22%) cases. Pelvic tilt was the only pre-operative predictive factor of this difference. CONCLUSION: Our study showed that anteversion of the acetabular component following THA using the anterior approach was greater than the recommended target value, and that substantial differences were observed in some patients when measured using two different measurement planes. If these results are confirmed by further studies, and considering that the anterior approach is intended to limit the incidence of dislocation, a new correlation study for each reference plane (anatomical and functional) will be necessary to define a 'safe zone' for use with the anterior approach. TAKE HOME MESSAGE: EOS imaging system is helpful in the pre-operative and post-operative radiological analysis of total hip arthroplasty.
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Artroplastia de Reemplazo de Cadera/métodos , Articulación de la Cadera/diagnóstico por imagen , Acetábulo/diagnóstico por imagen , Acetábulo/patología , Acetábulo/cirugía , Anciano , Estudios de Cohortes , Femenino , Fémur/diagnóstico por imagen , Fémur/patología , Prótesis de Cadera , Humanos , Masculino , Persona de Mediana Edad , Osteoartritis de la Cadera/cirugía , Posicionamiento del Paciente/métodos , Postura , Estudios Prospectivos , Dosis de Radiación , RadiografíaRESUMEN
BACKGROUND: The outcome of revision total hip arthroplasty (THA) for intra-pelvic cup protrusion is unclear. Hence, we conducted a large retrospective study to clarify the surgical strategy (hip lever arm and cup mechanical fixation) and the outcomes of reconstruction for severe intra-pelvic cup protrusion. HYPOTHESIS: We hypothesized that restoration of the anatomic hip centre in such acetabular revisions decreased the risk of recurrent loosening. MATERIAL AND METHODS: The study included 246 THA procedures (in 220 patients), with a follow-up of 5.2 ± 4.9 years (1-24.2) after the index surgery. Bone loss was estimated using the SOFCOT classification (grade III or IV in 80% of cases) and the Paprosky classification (IIIA or IIIB in 58% of cases). Quality of the reconstruction was assessed on X-rays according to the correction of the protrusion and position of the hip centre of rotation. RESULTS: After a clinical follow-up of at least 5 years, with a mean of 9.9 ± 4.1 years (5-24 years), the mean Postel-Merle d'Aubigné score was 14.2 ± 3.1 and the mean Harris Hip Score was 78.0 ± 18.7. Cup protrusion was partially or completely corrected in every case and cup position was normal in 27 (11%) cases. The centre of rotation was within 10mm of the physiological position in 158 (64.2%) cases, acceptable in 77 (31.3%) cases, ascended in 9 (3.7%) cases, and worsened in 1 (0.4%) case. Revision for cup or cup and femoral failures was required in 24 (9.8%) cases. Cumulative survival rates with cup loosening as the endpoint were 88.5% after 5 years, 79.9% after 10 years, and 63.9% at last follow-up at 13.6 years. DISCUSSION: Our hypothesis that restoration of anatomic hip centre decreased the risk of recurrent loosening was not verified: success or failure in restoring the normal centre of rotation did not correlate significantly with final cup status. Recurrent aseptic loosening was the cause of failure in 9.8% of cases. Ensuring long-term effective mechanical stability had a greater impact on global outcomes than restoring an ideal centre of rotation.
Asunto(s)
Artroplastia de Reemplazo de Cadera/efectos adversos , Articulación de la Cadera/cirugía , Inestabilidad de la Articulación/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Estudios de Seguimiento , Articulación de la Cadera/fisiopatología , Humanos , Inestabilidad de la Articulación/etiología , Inestabilidad de la Articulación/fisiopatología , Masculino , Persona de Mediana Edad , Falla de Prótesis , Rango del Movimiento Articular , Reoperación , Estudios Retrospectivos , Factores de Tiempo , Resultado del Tratamiento , Adulto JovenRESUMEN
Adsorbed vs. intercalated MCPA (4-chloro-2-methylphenoxyacetic acid) in highly hydrated clays taken as a soil model were clearly distinguished by 1H HR MAS NMR; adsorbed herbicide gave sharp signals indicating high mobility while intercalated herbicide gave very wide unresolved spectra due to its strong interaction with the solid matrix.
Asunto(s)
Ácido 2-Metil-4-clorofenoxiacético/análisis , Residuos de Plaguicidas/análisis , Contaminantes del Suelo/análisis , Espectroscopía de Resonancia Magnética , Modelos QuímicosRESUMEN
In vivo or in situ nuclear magnetic resonance (NMR) offers a powerful tool to study the degradation of xenobiotics by microorganisms. Most studies reported are based on the use of heteronuclei, and experiments with xenobiotics have been limited because specifically labeled xenobiotics are not commercially available, with the exception of (19)F and (31)P. (1)H NMR is, thus, of great interest in this area. To avoid problems caused by the presence of water and intrinsic metabolite signals, some studies were performed using a deuterated medium or specific detection of protons linked to the (13)C-(15)N enriched pattern. We report here the application of in situ (1)H NMR, performed directly on culture media, to study the metabolism of heterocyclic compounds. In this review, we show that a common pathway is involved in the biodegradation of morpholine, piperidine, and thiomorpholine by Mycobacterium aurum MO1 and Mycobacterium sp. RP1. In all cases, the first step is the cleavage of the C-N bond, which results in an amino acid. Thiomorpholine is first oxidized to sulfoxide before the opening of the ring. The second step is the deamination of the intermediate amino acid, which leads to the formation of a diacid. We have shown that the cleavage of the C-N bond and the oxidation of thiomorpholine are initiated by reactions involving a cytochrome P450.
RESUMEN
AIM: The aim of this study was to find and characterize a fungal or bacterial strain capable of metabolizing mesotrione, a new selective herbicide for control of broad-leaved weeds in maize. METHODS AND RESULTS: This strain was isolated from cloud water and showed close phylogenetic relationship with strains belonging to the Bacillus genus, based on 16S rRNA gene alignment. Kinetics of mesotrione degradation were monitored by high-performance liquid chromatography and in situ(1)H nuclear magnetic resonance spectroscopy at different concentrations. Mesotrione was completely biotransformed even at 5 mmol l(-1) concentration. 2-Amino-4-methylsulfonyl benzoic acid (AMBA) was identified as one of the metabolites, but was not the major one. CONCLUSIONS: This study reports the first rapid mesotrione biotransformation by a pure bacterial strain and the formation of several metabolites including AMBA. SIGNIFICANCE AND IMPACT OF THE STUDY: This bacterium isolated from cloud water is the first pure strain capable of rapidly degrading mesotrione.
Asunto(s)
Atmósfera , Bacillus/aislamiento & purificación , Bacillus/metabolismo , Ciclohexanonas/metabolismo , Herbicidas/metabolismo , Microbiología del Agua , Altitud , Bacillus/clasificación , Bacillus/genética , Biodegradación Ambiental , Francia , Genes de ARNr , ARN Ribosómico 16S/genéticaRESUMEN
In vivo or in situ nuclear magnetic resonance (NMR) offers a powerful tool to study the degradation of xenobiotics by microorganisms. Most studies reported are based on the use of heteronuclei, and experiments with xenobiotics have been limited because specifically labeled xenobiotics are not commercially available, with the exception of 19F and 31P. wn>1H NMR is, thus, of great interest in this area. To avoid problems caused by the presence of water and intrinsic metabolite signals, some studies were performed using a deuterated medium or specific detection of protons linked to the 13C-15N enriched pattern. We report here the application of in situ 1H NMR, performed directly on culture media, to study the metabolism of heterocyclic compounds. In this review, we show that a common pathway is involved in the biodegradation of morpholine, piperidine, and thiomorpholine by Mycobacterium aurum MO1 and Mycobacterium sp. RP1. In all cases, the first step is the cleavage of the C-N bond, which results in an amino acid. Thiomorpholine is first oxidized to sulfoxide before the opening of the ring. The second step is the deamination of the intermediate amino acid, which leads to the formation of a diacid. We have shown that the cleavage of the C-N bond and the oxidation of thiomorpholine are initiated by reactions involving a cytochrome P450.
Asunto(s)
Compuestos Heterocíclicos/metabolismo , Hidrógeno/metabolismo , Espectroscopía de Resonancia Magnética/métodos , Mycobacterium/metabolismo , Xenobióticos/metabolismo , Biodegradación Ambiental , Morfolinas/metabolismo , Piperidinas/metabolismoRESUMEN
One- and two-dimensional (1)H NMR spectroscopy were used to study the biotransformation of two dietary glucosinolates, sinigrin (SIN), and glucotropaeolin (GTL) by the human digestive microflora in vitro. The molecular structures of the new metabolites issued from the aglycone moiety of the glucosinolate were identified, and the modulation of carbon metabolism was studied by quantifying bacterial metabolites issued from the xenobiotic incubation in the presence or absence of a source of free glucose. Unambiguously and for the first time, it was shown that SIN and GTL were transformed quantitatively into allylamine and benzylamine, respectively. The comparison of the kinetics of transformation of SIN and GTL with and without glucose clearly showed that the presence of glucose did not modify either the nature of the metabolites or the rate of transformation of the glucosinolates (complete degradation within 30 h). The main end products of the glucose moiety of glucosinolates were characteristic of anaerobic carbon metabolism in the digestive tract (acetate, lactate, ethanol, propionate, formate, and butyrate) and similar to those released from free glucose. This work represents the first application of (1)H NMR spectroscopy to the study of xenobiotic metabolism by the human digestive microflora, demonstrating allyl- and benzylamine production from glucosinolates. Whether these amines are produced in vivo from dietary glucosinolates remains to be established. This would reduce the availability of other glucosinolate metabolites, notably cancer-protective isothiocyanates.
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Heces/química , Heces/microbiología , Glucosinolatos/metabolismo , Ácidos Hidroxámicos/metabolismo , Isotiocianatos , Resonancia Magnética Nuclear Biomolecular , Tioglucósidos/metabolismo , Adulto , Brassica , Sistema Digestivo/metabolismo , Sistema Digestivo/microbiología , Glucosinolatos/química , Humanos , Ácidos Hidroxámicos/química , Cinética , Resonancia Magnética Nuclear Biomolecular/métodos , Protones , Tioglucósidos/químicaRESUMEN
The biodegradation of benzothiazole and 2-hydroxybenzothiazole by two strains of Rhodococcus was monitored by reversed phase high-pressure liquid chromatography and by (1)H nuclear magnetic resonance (NMR). Both xenobiotics were biotransformed into a hydroxylated derivative of 2-hydroxybenzothiazole by these two strains. The chemical structure of this metabolite was determined by a new NMR methodology: long-range (1)H-(15)N heteronuclear shift correlation without any previous (15)N enrichment of the compound. This powerful NMR tool allowed us to assign the metabolite structure to 2,6-dihydroxybenzothiazole.
Asunto(s)
Rhodococcus/metabolismo , Tiazoles/química , Tiazoles/metabolismo , Benzotiazoles , Biodegradación Ambiental , Cromatografía Líquida de Alta Presión/métodos , Espectroscopía de Resonancia Magnética , Rhodococcus/crecimiento & desarrolloRESUMEN
In order to see if the biodegradative pathways for morpholine and thiomorpholine during degradation by Mycobacterium aurum MO1 could be generalized to other heterocyclic compounds, the degradation of piperidine by this strain was investigated by performing (1)H-nuclear magnetic resonance directly with the incubation medium. Ionspray mass spectrometry, performed without purification of the samples, was also used to confirm the structure of some metabolites during morpholine and thiomorpholine degradation. The results obtained with these two techniques suggested a general pathway for degradation of nitrogen heterocyclic compounds by M. aurum MO1. The first step of the degradative pathway is cleavage of the C---N bond; this leads formation of an intermediary amino acid, which is followed by deamination and oxidation of this amino acid into a diacid. Except in the case of thiodiglycolate obtained from thiomorpholine degradation, the dicarboxylates are completely mineralized by the bacterial cells. A comparison with previously published data showed that this pathway could be a general pathway for degradation by other strains of members of the genus Mycobacterium.
Asunto(s)
Morfolinas/metabolismo , Mycobacterium/metabolismo , Piperidinas/metabolismo , Biodegradación Ambiental , Medios de Cultivo , Espectroscopía de Resonancia Magnética , Espectrometría de Masas/métodos , Mycobacterium/crecimiento & desarrolloRESUMEN
Resting Mycobacterium aurum MO1 cells were incubated with morpholine, a waste from the chemical industry. The kinetics of biodegradation was monitored by using in situ nuclear magnetic resonance (NMR). The incubation medium was directly analyzed by 1H NMR. This technique allowed the unambiguous identification of two intermediates of the metabolic pathway involved in the biodegradation process, glycolate and 2-(2-aminoethoxy)acetate. The latter compound, which was not commercially available, was synthesized, in three steps, from 2-(2-aminoethoxy)ethanol. Quantitative analysis of the kinetics of degradation of morpholine was performed by integrating the signals of the different metabolites in 1H-NMR spectra. Morpholine was degraded within 10 h. The intermediates increased during the first 10 h and finally disappeared after 20 h incubation. Assays of degradation were also carried out with glycolate and ethanolamine, hypothetical intermediates of the morpholine degradation pathway. They were degraded within 4 and 8 h, respectively. Until now, no tool for direct detection of intermediates or even morpholine has been available, consequently, only hypothetical pathways have been proposed. The approach described here gives both qualitative and quantitative information about the metabolic routes used in morpholine degradation by M. aurum MO1. It could be used to investigate many biodegradative processes.
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Acetatos/análisis , Etilaminas/análisis , Glicolatos/análisis , Morfolinas/metabolismo , Mycobacterium/metabolismo , Acetatos/metabolismo , Biodegradación Ambiental , Etanol/análogos & derivados , Etanol/metabolismo , Etanolamina/análisis , Etanolamina/metabolismo , Etilaminas/metabolismo , Glicolatos/metabolismo , Hidrógeno/análisis , Residuos Industriales , Espectroscopía de Resonancia Magnética , Mycobacterium/crecimiento & desarrolloRESUMEN
A Mycobacterium strain (RP1) was isolated from a contaminated activated sludge collected in a wastewater treatment unit of a chemical plant. It was capable of utilizing morpholine and other heterocyclic compounds, such as pyrrolidine and piperidine, as the sole source of carbon, nitrogen, and energy. The use of in situ 1H nuclear magnetic resonance (1H NMR) spectroscopy allowed the determination of two intermediates in the biodegradative pathway, 2-(2-aminoethoxy)acetate and glycolate. The inhibitory effects of metyrapone on the degradative abilities of strain RP1 indicated the involvement of a cytochrome P-450 in the biodegradation of morpholine. This observation was confirmed by spectrophotometric analysis and 1H NMR. Reduced cell extracts from morpholine-grown cultures, but not succinate-grown cultures, gave rise to a carbon monoxide difference spectrum with a peak near 450 nm, which indicated the presence of a soluble cytochrome P-450. 1H NMR allowed the direct analysis of the incubation medium containing metyrapone, a specific inhibitor of cytochrome P-450. The inhibition of morpholine degradation was dependent on the morpholine/metyrapone ratio. The heme-containing monooxygenase was also detected in pyrrolidine- and piperidine-grown cultures. The abilities of different compounds to support strain growth or the induction of a soluble cytochrome P-450 were assayed. The results suggest that this enzyme catalyzes the cleavage of the C-N bond of the morpholine ring.
Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Morfolinas/metabolismo , Mycobacterium/metabolismo , Acetatos/análisis , Acetatos/metabolismo , Biodegradación Ambiental , Monóxido de Carbono/metabolismo , Etanolamina/metabolismo , Etilaminas/análisis , Etilaminas/metabolismo , Glicolatos/análisis , Glicolatos/metabolismo , Hidrógeno/análisis , Residuos Industriales , Espectroscopía de Resonancia Magnética , Metirapona/farmacología , Mycobacterium/aislamiento & purificación , Piperidinas/metabolismo , Pirrolidinas/metabolismo , Aguas del Alcantarillado/microbiología , Ácido Succínico/metabolismoRESUMEN
Spectrophotometric assays of Mycobacterium aurum MO1 cells extracts gave evidence of a soluble cytochrome P450, involved in the degradative pathway of morpholine, a waste product from the chemical industry. In order to get further information, the kinetics of the biodegradation of the sulfur analogue thiomorpholine was monitored by using in situ nuclear magnetic resonance (NMR). This technique allowed the identification of two intermediates: the sulfoxide of thiomorpholine resulting from S-oxidation and thiodiglycolic acid owing to ring cleavage. The S-oxidation (S-->SO) represents one of the well-known reactions catalyzed by cytochromes P450. The inhibitory effect of metyrapone, a cytochrome P450 inhibitor, on the thiomorpholine and morpholine degradative abilities of M. aurum MO1 confirmed the involvement of a cytochrome P450. These results and the decrease of the rate of formation of the first intermediate during the morpholine degradation, 2-(2-aminoethoxy) acetate, proved the key role of the cytochrome P450 in the early events of the biodegradation, i.e., in the C-N bond cleavage.