Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 39
Filtrar
Más filtros

Banco de datos
Tipo del documento
Intervalo de año de publicación
1.
Chem Rev ; 122(9): 8126-8180, 2022 05 11.
Artículo en Inglés | MEDLINE | ID: mdl-35234463

RESUMEN

Microorganisms have gained defense systems during the lengthy process of evolution over millions of years. Such defense systems can protect them from being attacked by invading species (e.g., CRISPR-Cas for establishing adaptive immune systems and nanopore-forming toxins as virulence factors) or enable them to adapt to different conditions (e.g., gas vesicles for achieving buoyancy control). These microorganism defense systems (MDS) have inspired the development of biosensors that have received much attention in a wide range of fields including life science research, food safety, and medical diagnosis. This Review comprehensively analyzes biosensing platforms originating from MDS for sensing and imaging biological analytes. We first describe a basic overview of MDS and MDS-inspired biosensing platforms (e.g., CRISPR-Cas systems, nanopore-forming proteins, and gas vesicles), followed by a critical discussion of their functions and properties. We then discuss several transduction mechanisms (optical, acoustic, magnetic, and electrical) involved in MDS-inspired biosensing. We further detail the applications of the MDS-inspired biosensors to detect a variety of analytes (nucleic acids, peptides, proteins, pathogens, cells, small molecules, and metal ions). In the end, we propose the key challenges and future perspectives in seeking new and improved MDS tools that can potentially lead to breakthrough discoveries in developing a new generation of biosensors with a combination of low cost; high sensitivity, accuracy, and precision; and fast detection. Overall, this Review gives a historical review of MDS, elucidates the principles of emulating MDS to develop biosensors, and analyzes the recent advancements, current challenges, and future trends in this field. It provides a unique critical analysis of emulating MDS to develop robust biosensors and discusses the design of such biosensors using elements found in MDS, showing that emulating MDS is a promising approach to conceptually advancing the design of biosensors.


Asunto(s)
Técnicas Biosensibles , Nanoporos , Ácidos Nucleicos , Sistemas CRISPR-Cas , Proteínas
2.
Extremophiles ; 25(4): 327-342, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-33993356

RESUMEN

Stromatolites are organo-sedimentary structures found principally in seas and saline lakes that contain sheets of sediments and minerals formed by layers of microbial communities, which trap sediments and induce the precipitation of minerals.A living stromatolite from the alkaline Laguna Interna in the Salar de Atacama was collected and one of the fragments was deposited in an experimental aquarium for 18 months. We used Illumina sequencing of PCR-amplified V4 regions of 16S rRNA genes from total extracted DNA to identify the microbial populations. The chemical structure was studied using X-Ray Diffraction (XRD) and bench chemical methods. We found that members belonging to the Proteobacteria, Planctomycetes, Chloroflexi and Bacteroidetes phyla dominated the bacterial communities of the living and aquarium cultured samples. The potential metabolic functionality of the prokaryotic community reveals that sulfur, nitrogen, methane and carbon fixation metabolism functions are present in the samples. This study is the first to provide new insights into the prokaryotic community composition from this unusual aquatic desert site. Further studies will be helpful to obtain a better understanding of the biotic and abiotic mechanisms residing in stromatolites from Laguna Interna, as well as to have better knowledge about the formation of these biosignatures.


Asunto(s)
Biodiversidad , Sedimentos Geológicos , Chile , ADN Bacteriano , Laboratorios , Filogenia , ARN Ribosómico 16S
3.
Curr Microbiol ; 77(8): 1429-1437, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32193606

RESUMEN

The Desert of Maine, not a real desert, is a 160,000 m2 tourist attraction of glacial silt which resembles a desert, surrounded by a pine forest in the state of Maine located in the northeastern USA. Though not a true desert, the soil of the Desert of Maine has a sandy texture with poor water-holding abilities, nutrient retention capabilities, and a relatively low pH value (pH 5.09). Samples from this site may be of interest to examine the bacterial diversity present on mineral sandy loam soils with an acidic pH, low concentrations of organic materials though surrounded by a pine forest, and compare it with true desert soil microbial populations. Two surface sand samples from the Desert of Maine were obtained, and pyrosequencing of PCR amplified 16S rRNA genes from total extracted DNA was used to assess bacterial diversity, community structure, and the relative abundance of major bacterial taxa. We found that the soil samples from the Desert of Maine displayed high levels of bacterial diversity, with a predominance of members belonging to the Proteobacteria and Actinobacteria phyla. Bacteria from the most abundant genus, Acidiphilium, represent 12.5% of the total 16S rDNA sequences. In total, 1394 OTUs were observed in the two samples, with 668 OTUs being observed in both samples. By comparing Desert of Maine bacterial populations with studies on similar soil environments, we found that the samples contained less Acidobacteria than soils from acid soil forests, and less Firmicutes plus more Proteobacteria than oligotrophic desert soils. Interestingly, our samples were found to be highly similar in their composition to an oak forest soil in France.


Asunto(s)
Bacterias/clasificación , Microbiota , Arena/microbiología , Microbiología del Suelo , Bacterias/aislamiento & purificación , Clima Desértico , Secuenciación de Nucleótidos de Alto Rendimiento , Concentración de Iones de Hidrógeno , Maine , Minerales , ARN Ribosómico 16S/genética
4.
Antonie Van Leeuwenhoek ; 112(3): 351-365, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30232678

RESUMEN

Salinity is an important environmental factor influencing microbial community composition. To better understand this influence, we determined the bacterial communities present in 17 different sites of brackish sediment (underwater) and soil (surface) samples from the Camargue region (Rhône river delta) in southern France during the fall of 2013 and 2014 using pyrosequencing of the V3-V4 regions of the 16S rRNA genes amplified by PCR. This region is known for abundant flora and fauna and, though saline, 30% of rice consumed in France is grown here. We found that bacterial abundance in 1 g of soil or sediment, calculated by qPCR, was higher in sediments than in surface soil samples. Members belonging to the Proteobacteria, Bacteroidetes, Chloroflexi and Firmicutes phyla dominated the bacterial communities of sediment samples, while members belonging to the Proteobacteria, Bacteroidetes, Gemmatimonadetes, Actinobacteria, Firmicutes and Acidobacteria phyla dominated the bacterial communities of the soil samples. The most abundant bacterial genera present in the saline sediments and soils from the Camargue belonged mostly to halophilic and sulphate reducing bacteria, suggesting that the Camargue may be a valuable system to investigate saline, yet agriculturally productive, sediment and soil microbial ecosystem.


Asunto(s)
Bacterias/clasificación , Bacterias/aislamiento & purificación , Biodiversidad , Estuarios , Sedimentos Geológicos/microbiología , Microbiología del Suelo , Bacterias/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Francia , Mar Mediterráneo , Metagenómica , Filogenia , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
5.
Scand J Gastroenterol ; 53(10-11): 1358-1363, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30394137

RESUMEN

OBJECTIVES: Evaluation of indeterminate biliary strictures remains challenging due to limited sensitivity of endoscopic tissue sampling. Biliary probe-based confocal laser endomicroscopy (pCLE) has shown promise to detect and exclude neoplasia. However, knowledge of whether individual inflammatory criteria are more prevalent in neoplasia compared to benign strictures is limited. The objective of this work is to improve diagnosis of neoplastic and inflammatory conditions using pCLE. MATERIALS AND METHODS: The charts of all patients who underwent pCLE at a single referral center between 2009 and 2015 were reviewed. ERCP reports were reviewed for eleven Miami and Paris criteria. Primary outcome was the identification of neoplasia by histopathology (defined as high-grade dysplasia and/or adenocarcinoma). To model predictors of neoplasia, we fit a binary regression model incorporating data from pCLE operating criteria, pCLE impression, and PSC status. RESULTS: 97 patients were identified. In the 27 patients with neoplasia, there was increasing number of Miami malignant criteria (Pearson r = 0.512, p < .001) while inflammatory criteria were less prevalent. 10% (5/51, p < .001) of patients with benign pCLE impression developed neoplasia, while 48% (22/46, p < .001) with suspicious pCLE impressions developed neoplasia. The binary regression model to predict neoplasia had a sensitivity of 83.3%, specificity of 92.5%, and overall accuracy 89.7%. CONCLUSIONS: Presence of malignant criteria and absence of certain inflammatory criteria are more prevalent in patients with neoplasia. Our model, which weights individual imaging components, shows impressive sensitivity and specificity over prior prognostic efforts. Prospective studies will be required to evaluate this model.


Asunto(s)
Neoplasias de los Conductos Biliares/diagnóstico , Conductos Biliares Intrahepáticos , Colangiocarcinoma/diagnóstico , Colangiopancreatografia Retrógrada Endoscópica/normas , Microscopía Confocal/normas , Adulto , Anciano , Neoplasias de los Conductos Biliares/complicaciones , Colangiocarcinoma/complicaciones , Constricción Patológica/diagnóstico , Constricción Patológica/etiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Guías de Práctica Clínica como Asunto , Curva ROC , Análisis de Regresión , Sensibilidad y Especificidad
6.
Retina ; 35(10): 2028-36, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25932560

RESUMEN

PURPOSE: Eyes fellow to nonischemic central retinal vein occlusion (CRVO) were examined for abnormalities, which might explain their increased risk for future occlusion, using adaptive optics scanning light ophthalmoscope fluorescein angiography. METHODS: Adaptive optics scanning light ophthalmoscope fluorescein angiography foveal microvascular densities were calculated. Nonperfused capillaries adjacent to the foveal avascular zone were identified. Spectral domain optical coherence tomography, ultrawide field fluorescein angiographies, and microperimetry were also performed. RESULTS: Ten fellow eyes of nine nonischemic CRVO and 1 nonischemic hemi-CRVO subjects and four affected eyes of three nonischemic CRVO and one nonischemic hemi-CRVO subjects were imaged. Ninety percent of fellow eyes and 100% of affected eyes demonstrated at least 1 nonperfused capillary compared with 31% of healthy eyes. Fellow eye microvascular density (35 ± 3.6 mm(-1)) was significantly higher than that of affected eyes (25 ± 5.2 mm(-1)) and significantly lower than that of healthy eyes (42 ± 4.2 mm(-1)). Compared with healthy controls, spectral domain optical coherence tomography thicknesses showed no significant difference, whereas microperimetry and 2/9 ultrawide field fluorescein angiography revealed abnormalities in fellow eyes. CONCLUSION: Fellow eye changes detectable on adaptive optics scanning light ophthalmoscope fluorescein angiography reflect subclinical pathology difficult to detect using conventional imaging technologies. These changes may help elucidate the pathogenesis of nonischemic CRVO and help identify eyes at increased risk of future occlusion.


Asunto(s)
Lateralidad Funcional/fisiología , Flujo Sanguíneo Regional/fisiología , Oclusión de la Vena Retiniana/diagnóstico , Oclusión de la Vena Retiniana/fisiopatología , Vasos Retinianos/patología , Adulto , Anciano , Capilares/patología , Femenino , Angiografía con Fluoresceína , Humanos , Isquemia/diagnóstico , Isquemia/fisiopatología , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Tomografía de Coherencia Óptica , Pruebas del Campo Visual , Campos Visuales/fisiología , Adulto Joven
7.
Retina ; 34(10): 2133-46, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24752010

RESUMEN

PURPOSE: To evaluate outer retinal structural abnormalities in patients with visual deficits after closed-globe blunt ocular trauma. METHODS: Nine subjects with visual complaints after closed-globe blunt ocular trauma were examined between 1 month after trauma and 6 years after trauma. Spectral domain optical coherence tomography was used to assess the outer retinal architecture, whereas adaptive optics scanning light ophthalmoscopy was used to analyze the photoreceptor mosaic integrity. RESULTS: Visual deficits ranged from central scotomas to decreased visual acuity. Spectral domain optical coherence tomography defects included focal foveal photoreceptor lesions, variable attenuation of the interdigitation zone, and mottling of the outer segment band, with one subject having normal outer retinal structure. Adaptive optics scanning light ophthalmoscopy revealed disruption of the photoreceptor mosaic in all subjects, variably manifesting as foveal focal discontinuities, perifoveal hyporeflective cones, and paracentral regions of selective cone loss. CONCLUSION: We observe persistent outer retinal disruption in subjects with visual complaints after closed-globe blunt ocular trauma, albeit to a variable degree. Adaptive optics scanning light ophthalmoscopy imaging allows the assessment of photoreceptor structure at a level of detail not resolvable using spectral domain optical coherence tomography or other current clinical imaging tools. Multimodal imaging seems to be useful in revealing the cause of visual complaints in patients after closed-globe blunt ocular trauma. Future studies are needed to better understand how photoreceptor structure changes longitudinally in response to various traumas.


Asunto(s)
Lesiones Oculares/patología , Retina/lesiones , Heridas no Penetrantes/patología , Accidentes de Tránsito , Adolescente , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Oftalmoscopía , Células Fotorreceptoras de Vertebrados/patología , Retina/patología , Tomografía de Coherencia Óptica , Trastornos de la Visión/patología , Agudeza Visual/fisiología , Adulto Joven
8.
Microb Ecol ; 66(4): 850-60, 2013 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-23963222

RESUMEN

Arid regions represent nearly 30 % of the Earth's terrestrial surface, but their microbial biodiversity is not yet well characterized. The surface sands of deserts, a subset of arid regions, are generally subjected to large temperature fluctuations plus high UV light exposure and are low in organic matter. We examined surface sand samples from the Taklamaken (China, three samples) and Gobi (Mongolia, two samples) deserts, using pyrosequencing of PCR-amplified 16S V1/V2 rDNA sequences from total extracted DNA in order to gain an assessment of the bacterial population diversity. In total, 4,088 OTUs (using ≥97 % sequence similarity levels), with Chao1 estimates varying from 1,172 to 2,425 OTUs per sample, were discernable. These could be grouped into 102 families belonging to 15 phyla, with OTUs belonging to the Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria phyla being the most abundant. The bacterial population composition was statistically different among the samples, though members from 30 genera were found to be common among the five samples. An increase in phylotype numbers with increasing C/N ratio was noted, suggesting a possible role in the bacterial richness of these desert sand environments. Our results imply an unexpectedly large bacterial diversity residing in the harsh environment of these two Asian deserts, worthy of further investigation.


Asunto(s)
Bacterias/aislamiento & purificación , Biodiversidad , Sedimentos Geológicos/microbiología , Bacterias/clasificación , Bacterias/genética , China , Clima Desértico , Datos de Secuencia Molecular , Filogenia , Dióxido de Silicio/análisis
9.
Antonie Van Leeuwenhoek ; 103(6): 1329-41, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23559041

RESUMEN

Arid zones cover over 30 % of the Earth's continental surface. In order to better understand the role of microbes in this type of harsh environment, we isolated and characterized the bacteriophages from samples of the surface sand of the Mesquite Flats region via electron microscopy and DNA sequencing of a select number of cloned phage DNAs. An electron microscopic analysis of the recovered virus-like particles revealed at least 11 apparently different morphotypes sharing structural characteristics of the Caudoviridae family of tailed phages. We found that 36 % of the sequences contained no significant identity (e-value >10(-3)) with sequences in the databases. Pilot sequencing of cloned 16S rRNA genes identified Bacteroidetes and Proteobacteria as the major bacterial groups present in this severe environment. The majority of the 16S rDNA sequences from the total (uncultured) bacterial population displayed ≤96 % identity to 16S rRNA genes in the database, suggesting an unexplored bacterial population likely adapted to a desert environment. In addition, we also isolated and identified 38 cultivable bacterial strains, the majority of which belonged to the genus Bacillus. Mitomycin-C treatment of the cultivable bacteria demonstrated that the vast majority (84 %) contained at least one SOS-inducible prophage.


Asunto(s)
Bacillus , Bacteroidetes , Caudovirales , Proteobacteria , Microbiología del Suelo , Bacillus/clasificación , Bacillus/aislamiento & purificación , Bacillus/virología , Bacteroidetes/clasificación , Bacteroidetes/aislamiento & purificación , Bacteroidetes/virología , Secuencia de Bases , Biodiversidad , California , Caudovirales/clasificación , Caudovirales/genética , Caudovirales/aislamiento & purificación , ADN Bacteriano/genética , ADN Viral/genética , Clima Desértico , Técnicas de Amplificación de Ácido Nucleico , Filogenia , Proteobacteria/clasificación , Proteobacteria/aislamiento & purificación , Proteobacteria/virología , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Estados Unidos
10.
Appl Environ Microbiol ; 78(2): 481-92, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22081576

RESUMEN

Degradation of the quorum-sensing (QS) signals known as N-acylhomoserine lactones (AHL) by soil bacteria may be useful as a beneficial trait for protecting crops, such as potato plants, against the worldwide pathogen Pectobacterium. In this work, analytical chemistry and microbial and molecular approaches were combined to explore and compare biostimulation of native and introduced AHL-degrading Rhodococcus erythropolis populations in the rhizosphere of potato plants cultivated in farm greenhouses under hydroponic conditions. We first identified gamma-heptalactone (GHL) as a novel biostimulating agent that efficiently promotes plant root colonization by AHL-degrading R. erythropolis population. We also characterized an AHL-degrading biocontrol R. erythropolis isolate, R138, which was introduced in the potato rhizosphere. Moreover, root colonization by AHL-degrading bacteria receiving different combinations of GHL and R138 treatments was compared by using a cultivation-based approach (percentage of AHL-degrading bacteria), pyrosequencing of PCR-amplified rrs loci (total bacterial community), and quantitative PCR (qPCR) of the qsdA gene, which encodes an AHL lactonase in R. erythropolis. Higher densities of the AHL-degrading R. erythropolis population in the rhizosphere were observed when GHL treatment was associated with biocontrol strain R138. Under this condition, the introduced R. erythropolis population displaced the native R. erythropolis population. Finally, chemical analyses revealed that GHL, gamma-caprolactone (GCL), and their by-products, gamma-hydroxyheptanoic acid and gamma-hydroxycaproic acid, rapidly disappeared from the rhizosphere and did not accumulate in plant tissues. This integrative study highlights biostimulation as a potential innovative approach for improving root colonization by beneficial bacteria.


Asunto(s)
Acil-Butirolactonas/metabolismo , Percepción de Quorum , Rhodococcus/aislamiento & purificación , Rhodococcus/fisiología , Técnicas Bacteriológicas , Técnicas de Química Analítica , Raíces de Plantas/microbiología , Rizosfera , Rhodococcus/genética , Rhodococcus/crecimiento & desarrollo , Análisis de Secuencia de ADN , Solanum tuberosum/microbiología
11.
Appl Microbiol Biotechnol ; 91(3): 635-44, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21494865

RESUMEN

The screening of environmental DNA metagenome libraries for functional activities can provide an important source of new molecules and enzymes. In this study, we identified 17 potential protease-producing clones from two metagenomic libraries derived from samples of surface sand from the Gobi and Death Valley deserts. Two of the proteases, DV1 and M30, were purified and biochemically examined. These two proteases displayed a molecular mass of 41.5 kDa and 45.7 kDa, respectively, on SDS polyacrylamide gels. Alignments with known protease sequences showed less than 55% amino acid sequence identity. These two serine proteases appear to belong to the subtilisin (S8A) family and displayed several unique biochemical properties. Protease DV1 had an optimum pH of 8 and an optimal activity at 55°C, while protease M30 had an optimum pH >11 and optimal activity at 40°C. The properties of these enzymes make them potentially useful for biotechnological applications and again demonstrate that metagenomic approaches can be useful, especially when coupled with the study of novel environments such as deserts.


Asunto(s)
Metagenómica , Consorcios Microbianos/genética , Serina Proteasas/genética , Serina Proteasas/aislamiento & purificación , Dióxido de Silicio/análisis , Secuencia de Aminoácidos , Secuencia de Bases , Biomasa , California , China , Clima Desértico , Electroforesis en Gel de Poliacrilamida , Biblioteca de Genes , Mongolia , Nevada , Análisis de Secuencia por Matrices de Oligonucleótidos , Filogenia , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/genética , Alineación de Secuencia , Estados Unidos
12.
Microbiol Res ; 244: 126664, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33359841

RESUMEN

Desert-like areas located in the eastern portion of the state of Utah (USA) have geographic features that can resemble the surface of the planet Mars, characterized by red-colored hills, soils and sandstones. We examined the bacterial biodiversity of surface soil samples from several sites from the Colorado Plateau Desert in eastern Utah using pyrosequencing of PCR amplified bacterial 16S rRNA genes from total extracted soil DNA. The sample sites cover the Great Basin, Goblin Valley State Park and nearby regions on the Colorado Plateau. We also examined several physicochemical parameters of the soil samples to investigate any possible correlations between bacterial community structure and environmental drivers. The predominant bacterial phyla present in the samples were found to belong to members of the Proteobacteria, Actinobacteria, Bacteroidetes, and Gemmatimonadetes. The most abundant genera in our samples were found to belong to the Cesiribacter, Lysobacter, Adhaeribacter, Microvirga and Pontibacter genera. We found that the relative abundance of Proteobacteria and Gemmatimonadetes were significantly correlated with soil pH and a low concentration of organic matter, suggesting that, in these relatively high-altitude desert soils, these two parameters may be of primary importance to influence bacterial community composition.


Asunto(s)
Bacterias/aislamiento & purificación , Microbiología del Suelo , Bacterias/clasificación , Bacterias/genética , Colorado , ADN Bacteriano/genética , Microbiota , Filogenia , ARN Ribosómico 16S/genética , Suelo/química , Utah
13.
Virol J ; 7: 163, 2010 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-20637121

RESUMEN

BACKGROUND: Bacteriophage classification is mainly based on morphological traits and genome characteristics combined with host information and in some cases on phage growth lifestyle. A lack of molecular tools can impede more precise studies on phylogenetic relationships or even a taxonomic classification. The use of methods to analyze genome sequences without the requirement for homology has allowed advances in classification. RESULTS: Here, we proposed to use genome sequence signature to characterize bacteriophages and to compare them to their host genome signature in order to obtain host-phage relationships and information on their lifestyle. We analyze the host-phage relationships in the four most representative groups of Caudoviridae, the dsDNA group of phages. We demonstrate that the use of phage genomic signature and its comparison with that of the host allows a grouping of phages and is also able to predict the host-phage relationships (lytic vs. temperate). CONCLUSIONS: We can thus condense, in relatively simple figures, this phage information dispersed over many publications.


Asunto(s)
Bacterias/virología , Bacteriófagos/crecimiento & desarrollo , Bacteriófagos/genética , Evolución Molecular , Genómica , Interacciones Huésped-Patógeno , Bacterias/genética , Bacteriófagos/clasificación , Mapeo Cromosómico , Genoma Viral , Filogenia , Especificidad de la Especie
14.
J Ind Microbiol Biotechnol ; 37(2): 117-28, 2010 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19908076

RESUMEN

We examined the variations of bacterial populations in treated drinking water prior to and after the final chlorine disinfection step at two different surface water treatment plants. For this purpose, the bacterial communities present in treated water were sampled after granular activated carbon (GAC) filtration and chlorine disinfection from two drinking water treatment plants supplying the city of Paris (France). Samples were analyzed after genomic DNA extraction, polymerase chain reaction (PCR) amplification, cloning, and sequencing of a number of 16S ribosomal RNA (rRNA) genes. The 16S rDNA sequences were clustered into operational taxonomic units (OTUs) and the OTU abundance patterns were obtained for each sample. The observed differences suggest that the chlorine disinfection step markedly affects the bacterial community structure and composition present in GAC water. Members of the Alphaproteobacteria and Betaproteobacteria were found to be predominant in the GAC water samples after phylogenetic analyses of the OTUs. Following the chlorine disinfection step, numerous changes were observed, including decreased representation of Proteobacteria phylotypes. Our results indicate that the use of molecular methods to investigate changes in the abundance of certain bacterial groups following chlorine-based disinfection will aid in further understanding the bacterial ecology of drinking water treatment plants (DWTPs), particularly the disinfection step, as it constitutes the final barrier before drinking water distribution to the consumer's tap.


Asunto(s)
Bacterias/efectos de los fármacos , Cloro/farmacología , Agua Dulce/microbiología , Microbiología del Agua , Purificación del Agua , Abastecimiento de Agua , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Biodiversidad , Carbón Orgánico , Cloruros/farmacología , ADN Bacteriano/genética , ADN Ribosómico/análisis , Desinfección , Ecosistema , Compuestos Férricos/farmacología , Filtración , Agua Dulce/química , Paris , Filogenia , ARN Ribosómico 16S/genética , Purificación del Agua/métodos
15.
J Bacteriol ; 191(12): 3763-71, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19376864

RESUMEN

The primary sigma factor of Staphylococcus aureus, sigma(SA), regulates the transcription of many genes, including several essential genes, in this bacterium via specific recognition of exponential growth phase promoters. In this study, we report the existence of a novel staphylococcal phage G1-derived growth inhibitory polypeptide, referred to as G1ORF67, that interacts with sigma(SA) both in vivo and in vitro and regulates its activity. Delineation of the minimal domain of sigma(SA) that is required for its interaction with G1ORF67 as amino acids 294 to 360 near the carboxy terminus suggests that the G1 phage-encoded anti-sigma factor may occlude the -35 element recognition domain of sigma(SA). As would be predicted by this hypothesis, the G1ORF67 polypeptide abolished both RNA polymerase core-dependent binding of sigma(SA) to DNA and sigma(SA)-dependent transcription in vitro. While G1ORF67 profoundly inhibits transcription when expressed in S. aureus cells in mode of action studies, our finding that G1ORF67 was unable to inhibit transcription when expressed in Escherichia coli concurs with its inability to inhibit transcription by the E. coli holoenzyme in vitro. These features demonstrate the selectivity of G1ORF67 for S. aureus RNA polymerase. We predict that G1ORF67 is one of the central polypeptides in the phage G1 strategy to appropriate host RNA polymerase and redirect it to phage reproduction.


Asunto(s)
Regulación hacia Abajo , Péptidos/metabolismo , Factor sigma/metabolismo , Fagos de Staphylococcus/metabolismo , Staphylococcus aureus/genética , Transcripción Genética , Proteínas Virales/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , ARN Polimerasas Dirigidas por ADN/genética , ARN Polimerasas Dirigidas por ADN/metabolismo , Regulación Bacteriana de la Expresión Génica , Péptidos/genética , Unión Proteica , Estructura Terciaria de Proteína , Factor sigma/química , Factor sigma/genética , Fagos de Staphylococcus/genética , Staphylococcus aureus/metabolismo , Staphylococcus aureus/virología , Proteínas Virales/genética
16.
Viruses ; 11(1)2019 01 10.
Artículo en Inglés | MEDLINE | ID: mdl-30634666

RESUMEN

Enterococcus faecalis is an opportunistic pathogen that has emerged as a major cause of nosocomial infections worldwide. Many clinical strains are indeed resistant to last resort antibiotics and there is consequently a reawakening of interest in exploiting virulent phages to combat them. However, little is still known about phage receptors and phage resistance mechanisms in enterococci. We made use of a prophageless derivative of the well-known clinical strain E. faecalis V583 to isolate a virulent phage belonging to the Picovirinae subfamily and to the P68 genus that we named Idefix. Interestingly, most isolates of E. faecalis tested-including V583-were resistant to this phage and we investigated more deeply into phage resistance mechanisms. We found that E. faecalis V583 prophage 6 was particularly efficient in resisting Idefix infection thanks to a new abortive infection (Abi) mechanism, which we designated Abiα. It corresponded to the Pfam domain family with unknown function DUF4393 and conferred a typical Abi phenotype by causing a premature lysis of infected E. faecalis. The abiα gene is widespread among prophages of enterococci and other Gram-positive bacteria. Furthermore, we identified two genes involved in the synthesis of the side chains of the surface rhamnopolysaccharide that are important for Idefix adsorption. Interestingly, mutants in these genes arose at a frequency of ~10-4 resistant mutants per generation, conferring a supplemental bacterial line of defense against Idefix.


Asunto(s)
Bacteriófagos/patogenicidad , Enterococcus faecalis/genética , Enterococcus faecalis/virología , Podoviridae/patogenicidad , Bacteriófagos/aislamiento & purificación , Genoma Viral , Fenotipo , Profagos/genética , Aguas del Alcantarillado/virología , Virulencia , Secuenciación Completa del Genoma
17.
Appl Environ Microbiol ; 74(6): 1954-8, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18245262

RESUMEN

A tributyltin (TBT) luxAB transcriptional fusion in Escherichia coli revealed that a TBT-activated promoter is located upstream of two cotranscribed orphan genes, ygaV and ygaP. We demonstrate that transcription from the promoter upstream of ygaVP is constitutive in a ygaVP mutant, suggesting that YgaV is an autoregulated, TBT-inducible repressor.


Asunto(s)
Proteínas de Escherichia coli/genética , Escherichia coli/genética , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Operón , Proteínas Represoras/genética , Compuestos de Trialquiltina/farmacología , Ácido Anhídrido Hidrolasas/genética , Secuencia de Aminoácidos , Datos de Secuencia Molecular , Mutación , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas/genética , Homología de Secuencia de Aminoácido , Transcripción Genética/efectos de los fármacos
18.
J Microbiol ; 46(4): 364-72, 2008 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-18758725

RESUMEN

Bacteria and their viruses (called bacteriophages, or phages), have been found in virtually every ecological niche on Earth. Arid regions, including their most extreme form called deserts, represent the single largest ecosystem type on the Earth's terrestrial surface. The Namib desert is believed to be the oldest (80 million years) desert. We report here an initial analysis of bacteriophages isolated from the Namib desert using a combination of electron microscopy and genomic approaches. The virus-like particles observed by electron microscopy revealed 20 seemingly different phage-like morphologies and sizes belonging to the Myoviridae and Siphoviridae families of tailed phages. Pulsed-field gel electrophoresis revealed a majority of phage genomes of 55-65 kb in length, with genomes of approximately 200, 300, and 350 kb also observable. Sample sequencing of cloned phage DNA fragments revealed that approximately 50% appeared to be of bacterial origin. Of the remaining DNA sequences, approximately 50% displayed no significant match to any sequence in the databases. The majority of the 16S rDNA sequences amplified from DNA extracted from the sand displayed considerable (94-98%) homology to members of the Firmicutes, and in particular to members of the genus Bacillus, though members of the Bacteroidetes, Planctomycetes, Chloroflexi, and delta-Proteobacteria groups were also observed.


Asunto(s)
Bacterias/aislamiento & purificación , Bacteriófagos/aislamiento & purificación , Clima Desértico , Sedimentos Geológicos/microbiología , África , Bacterias/clasificación , Bacterias/genética , Tipificación de Bacteriófagos , Bacteriófagos/genética , Bacteriófagos/ultraestructura , ADN Bacteriano/genética , ADN Ribosómico/genética , ADN Viral/genética , Microscopía Electrónica , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 16S/genética
19.
Chem Sci ; 9(42): 8046-8055, 2018 Nov 14.
Artículo en Inglés | MEDLINE | ID: mdl-30542553

RESUMEN

MicroRNAs (miRNAs) play an important role in cellular functions and in the development and progression of cancer. Precise quantification of endogenous miRNAs from different clinical patient and control samples combined with a one-to-one comparison to standard technologies is a challenging but necessary endeavor that is largely neglected by many emerging fluorescence technologies. Here, we present a simple, precise, sensitive, and specific ratiometric assay for absolute quantification of miRNAs. Isothermally amplified time-gated Förster resonance energy transfer (TG-FRET) between Tb donors and dye acceptors resulted in miRNA assays with single-nucleotide variant specificity and detection limits down to 4.2 ± 0.5 attomoles. Quantification of miR-21 from human tissues and plasma samples revealed the relevance for breast and ovarian cancer diagnostics. Analysis of miR-132 and miR-146a from acute monocytic leukemia cells (THP-1) demonstrated the broad applicability to different miRNAs and other types of clinical samples. Direct comparison to the gold standard RT-qPCR showed advantages of amplified TG-FRET concerning precision and specificity when quantifying low concentrations of miRNAs as required for diagnostic applications. Our results demonstrate that a careful implementation of rolling circle amplification and TG-FRET into one straightforward nucleic acid detection method can significantly advance the possibilities of miRNA-based cancer diagnostics and research.

20.
Nat Biotechnol ; 22(2): 185-91, 2004 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-14716317

RESUMEN

Over evolutionary time bacteriophages have developed unique proteins that arrest critical cellular processes to commit bacterial host metabolism to phage reproduction. Here, we apply this concept of phage-mediated bacterial growth inhibition to antibiotic discovery. We sequenced 26 Staphylococcus aureus phages and identified 31 novel polypeptide families that inhibited growth upon expression in S. aureus. The cellular targets for some of these polypeptides were identified and several were shown to be essential components of the host DNA replication and transcription machineries. The interaction between a prototypic pair, ORF104 of phage 77 and DnaI, the putative helicase loader of S. aureus, was then used to screen for small molecule inhibitors. Several compounds were subsequently found to inhibit both bacterial growth and DNA synthesis. Our results suggest that mimicking the growth-inhibitory effect of phage polypeptides by a chemical compound, coupled with the plethora of phages on earth, will yield new antibiotics to combat infectious diseases.


Asunto(s)
Proteínas Bacterianas/metabolismo , Sistemas de Liberación de Medicamentos/métodos , Proteoma/metabolismo , Fagos de Staphylococcus/metabolismo , Staphylococcus aureus/metabolismo , Staphylococcus aureus/virología , Proteínas Virales/metabolismo , Antibacterianos/uso terapéutico , Infecciones Bacterianas/tratamiento farmacológico , Infecciones Bacterianas/metabolismo , Infecciones Bacterianas/virología , Bacteriófagos/metabolismo , Diseño de Fármacos , Perfilación de la Expresión Génica/métodos , Genoma Viral , Humanos , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/metabolismo , Infecciones Estafilocócicas/virología
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA