Large scale automated phylogenomic analysis of bacterial isolates and the Evergreen Online platform.

Public health authorities whole-genome sequence thousands of isolates each month for microbial diagnostics and surveillance of pathogenic bacteria. The computational methods have not kept up with the deluge of data and the need for real-time results. We have therefore created a bioinformatics pipeline for rapid subtyping and continuous phylogenomic analysis of bacterial samples, suited for large-scale surveillance. The data is divided into sets by mapping to reference genomes, then consensus sequences are generated. Nucleotide based genetic distance is calculated between the sequences in each set, and isolates are clustered together at 10 single-nucleotide polymorphisms. Phylogenetic trees are inferred from the non-redundant sequences and the clustered isolates are added back. The method is accurate at grouping outbreak strains together, while discriminating them from non-outbreak strains. The pipeline is applied in Evergreen Online, which processes publicly available sequencing data from foodborne bacterial pathogens on a daily basis, updating phylogenetic trees as needed.

A Bacterial Analysis Platform: An Integrated System for Analysing Bacterial Whole Genome Sequencing Data for Clinical Diagnostics and Surveillance.

Recent advances in whole genome sequencing have made the technology available for routine use in microbiological laboratories. However, a major obstacle for using this technology is the availability of simple and automatic bioinformatics tools. Based on previously published and already available web-based tools we developed a single pipeline for batch uploading of whole genome sequencing data from multiple bacterial isolates. The pipeline will automatically identify the bacterial species and, if applicable, assemble the genome, identify the multilocus sequence type, plasmids, virulence genes and antimicrobial resistance genes. A short printable report for each sample will be provided and an Excel spreadsheet containing all the metadata and a summary of the results for all submitted samples can be downloaded. The pipeline was benchmarked using datasets previously used to test the individual services. The reported results enable a rapid overview of the major results, and comparing that to the previously found results showed that the platform is reliable and able to correctly predict the species and find most of the expected genes automatically. In conclusion, a combined bioinformatics platform was developed and made publicly available, providing easy-to-use automated analysis of bacterial whole genome sequencing data. The platform may be of immediate relevance as a guide for investigators using whole genome sequencing for clinical diagnostics and surveillance. The platform is freely available at: https://cge.cbs.dtu.dk/services/CGEpipeline-1.1 and it is the intention that it will continue to be expanded with new features as these become available.

Amperometric inhibition biosensors based on horseradish peroxidase and gold sononanoparticles immobilized onto different electrodes for cyanide measurements.

New biosensors based on inhibition for the detection of cyanide and the comparison of the analytical performances of nine enzyme biosensor designs by using three different electrodes: Sonogel-Carbon, glassy carbon and gold electrodes were discussed. Three different horseradish peroxidase immobilization procedures with and without gold sononanoparticles were studied. The amperometric measurements were performed at an applied potential of -0.15V vs. Ag/AgCl in 50mM sodium acetate buffer solution pH=5.0. The apparent kinetic parameters (Kmapp, Vmaxapp) of immobilized HRP were calculated in the absence of inhibitor (cyanide) by using caffeic acid, hydroquinone, and catechol as substrates. The presence of gold sononanoparticles enhanced the electron transfer reaction and improved the analytical performance of the biosensors. The HRP kinetic interactions reveal non-competitive binding of cyanide with an apparent inhibition constant (Ki) of 2.7µM and I50 of 1.3µM. The determination of cyanide can be achieved in a dynamic range of 0.1-58.6µM with a detection limit of 0.03µM which is lower than those reported by previous studies. Hence this biosensing methodology can be used as a new promising approach for detecting cyanide.

Recent advances in graphite powder-based electrodes.

Graphite powder-based electrodes have the electrochemical performance of quasi-noble metal electrodes with intrinsic advantages related to the possibility of modification to enhance selectivity and their easily renewable surface, with no need for hazardous acids or bases for their cleaning. In contrast with commercial electrodes, for example screen-printed or sputtered-chip electrodes, graphite powder-based electrodes can also be fabricated in any laboratory with the form and characteristics desired. They are also readily modified with advanced materials, with relatively high reproducibility. All these characteristics make them a very interesting option for obtaining a large variety of electrodes to resolve different kinds of analytical problems. This review summarizes the state-of-the-art, advantages, and disadvantages of graphite powder-based electrodes in electrochemical analysis in the 21st century. It includes recent trends in carbon paste electrodes, devoting special attention to the use of emergent materials as new binders and to the development of other composite electrodes. The most recent advances in the use of graphite powder-modified sol-gel electrodes are also described. The development of sonogel-carbon electrodes and their use in electrochemical sensors and biosensors is included. These materials extend the possibilities of applications, especially for industrial technology-transfer purposes, and their development could affect not only electroanalytical green chemistry but other interesting areas also, for example catalysis and energy conversion and storage.

Use of a Sonogel-Carbon electrode modified with bentonite for the determination of diazepam and chlordiazepoxide hydrochloride in tablets and their metabolite oxazepam in urine.

Sonogel-Carbon electrode (SngCE) modified with bentonite (BENT) shows an interesting alternative electrode to be used in the determination of 1,4-benzodiazepines by square wave adsorptive cathodic stripping voltammetry (SWAdCSV). Diazepam (DZ) and chlordiazepoxide hydrochloride (CPZ), were determined using SngCE modified by 5% BENT. An electrochemical study of different parameters (such as pH, buffer type, ionic strength, accumulation potential, scan rate, and accumulation time) which affect the determination of DZ and CPZ is reported. Linear concentration ranges of 0.028-0.256 µg mL(-1) DZ (r=0.9997) and 0.034-0.302 µg mL(-1) CPZ (r=0.9997) are successfully obtained after an accumulation time of 60s. The quantification and detection limits were calculated to be 14.0 and 4.0 ng mL(-1) for DZ, and 16.0 and 5.0 ng mL(-1) for CPZ, respectively. The surface of the proposed electrode was characterized by scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDAX). The developed method was applied to the analysis of commercially available tablets and human urine real samples. Analysis was performed with better precision, very low detection limits, and faster than previously reported voltammetric techniques.

El color del miedo bajo el desorden del paisaje urbano en la ciudad de México y la zona metropolitana / The color of fear under the disorder of urban lanscape in Mexico City and the metropolitan zone / A cor do medo sob a desorganização da paisagem urbana em Cidade de México e da região metropolitana

Se presentan los resultados de un estudio exploratorio realizado en el 2008, sobre la percepción de la inseguridad en los pobladores de la Ciudad de México y la zona metropolitana, en el cual se utilizó la técnica del uso del color para relacionar la inseguridad con el espacio urbano, con el propósito de mostrar que los miedos no solo orbitan en las inmediaciones de las experiencias cotidianas, sino que, por el contrario, se alimentan de imaginarios sociales y mitos urbanos. Con estas líneas se pretende explorar la percepción de la inseguridad desde otras latitudes, despegándose de los estudios tradicionales de victimización, que si bien han servido para estudiar la criminalidad en México, como alternativa a las estadísticas oficiales, son apenas unos cuantos

Results are shown of an exploratory study carried out in 2008 about the perception of insecurity among the residents of Mexico City and the metropolitan zone; the use of color as a technique leading to relate insecurity to the urban space was chosen in order to show that fears do not orbit around the vicinities of daily experiences but, on the contrary, they are nurtured by social imaginaries and urban myths. These lines are intended to explore the perception of insecurity from other latitudes, straying away from traditional victimization studies that, although they have served to study criminality in Mexico, are still very few

Apresentam-se os resultados de um estudo exploratório realizado em 2008 sobre a percepção de insegurança nos moradores de Cidade de México e da zona metropolitana, no qual se utilizou a técnica do uso de cores para relacionar a insegurança com o espaço urbano, a fim de mostrar que os medos não só orbitam nas imediações das experiências cotidianas, mas, ao contrário, alimentam-se dos imaginários sociais e mitos urbanos. Estas linhas têm a intenção de explorar a percepção da insegurança de outras latitudes, separando-se dos estudos tradicionais de vitimização, que apesar de ter sido usados para estudar o crime no México, como uma alternativa para as estatísticas oficiais, são apenas alguns exemplos

1-Furoylthiourea-Sonogel-Carbon electrodes: structural and electrochemical characterization.

A new type of sensor based on Sonogel-Carbon materials modified with 1-(2-furoyl)-3-(1-naphthyl)thiourea is presented to be used as an amperometric sensor for metals. Different percentages of modifier were tested in order to optimise the structural and mechanical properties, as well as the electrochemical behaviour. Scanning electron microscopy (SEM), X-ray energy dispersive spectroscopy (EDS), Fourier transform-infrared spectroscopy (FTIR), single crystal X-ray diffraction (XRD) and powder XRD were used for the structural characterization of these electrodes. The 1-(2-furoyl)-3-(1-naphthyl)thiourea did not modify significantly the structural and mechanical properties of the Sonogel-Carbon electrodes, being similar to other modifications carried out previously in these materials. For the study of the electrochemical response, anodic stripping differential pulse voltammetry (ASDPV) was employed, optimising other parameters of measurement such as pH of the buffer, potential and accumulation time and pulse amplitude. The electrochemical response of the modified electrodes improved significantly with respect to the non-modified electrode, giving good signals and acceptable detection limit.

Sonogel-carbon electrode based on hemin for detection of superoxide.

The Sonogel electrode was immersed in hemin solution to obtain the hemin-Sonogel-modified electrode by cyclic voltammetry (CV). It was applied to detect superoxide radicals produced by xanthine oxidase catalyzed hypoxanthine oxidation. The characterization of hemin-Sonogel was modified by impedance and CV.

Alumina sol-gel/sonogel-carbon electrode based on acetylcholinesterase for detection of organophosphorus pesticides.

Two new amperometric biosensors based on immobilization of acetylcholinesterase on a sonogel-carbon electrode for detection of organophosphorous compounds are proposed. The electrodes were prepared applying high-energy ultrasounds directly to the precursors. The first biosensor was obtained by simple entrapping acetylcholinesterase in Al(2)O(3) sol-gel matrix on the sonogel-carbon. The second biosensor was produced in a sandwich configuration. Its preparation involved adsorption of the enzyme and modification via a polymeric membrane such as polyethylene glycol and the ion-exchanger Nafion. The optimal enzyme loading was found to be 0.7 mIU. Both biosensors showed optimal activity in 0.2 M phosphate buffer, pH 7.0, at an operating potential of 210 mV. The detection limit achieved for chlorpyriphos-ethyl-oxon was 2.5x10(-10)M at a 10-min incubation time.

Investigation of biosensor signal bioamplification: comparison of direct electrochemistry phenomena of individual Laccase, and dual Laccase-Tyrosinase copper enzymes, at a Sonogel-Carbon electrode.

Direct electrochemistry of Trametes versicolor Laccase (LAC) was found at a Sonogel-Carbon electrode. The bioamplification, performed by dual immobilization of this enzyme and Mushroom Tyrosinase (TYR), of the bio-electrocatalytic reduction of O(2) was investigated. The calculated alpha transfer coefficients were 0.64 and 0.67, and the heterogeneous electron-transfer rate constants were 6.19 and 8.52 s(-1), respectively, for the individual LAC and dual LAC-TYR-based Nafion/Sonogel-Carbon bio-electrodes. The responses of the dual enzymes electrode to polyphenols were stronger than those of the individual LAC or TYR biosensors. Hypotheses are offered about the mechanism of bioamplification. The surfaces of the biosensors were also characterized by AFM.

A comparison of three amperometric phenoloxidase-Sonogel-Carbon based biosensors for determination of polyphenols in beers.

Three phenoloxidases based biosensors were successfully developed using as electrochemical transducer a new type of electrode recently developed by our group: the "Sonogel-Carbon electrode". The employed enzymes were Trametes versicolor laccase (Lac), Mushroom tyrosinase (Tyr), and Horseradish peroxidase (HRP). Immobilization step was accomplished by doping the electrode surface with a mixture of the individual enzyme and Nafion ion exchanger as additive-protective. The biosensor responses, optimized in beer real samples, were evaluated for five individual polyphenols. It was found that the developed biosensors were sensitive to nanomolar concentrations of the tested polyphenols. As example, the limit of detection, sensitivity, and response linear range for caffeic acid for Nafion-Lac/Sonogel-Carbon biosensor were 0.06µmolL(-1), 99.6nAµmol(-1)L, and 0.04-2µmolL(-1), respectively. The stability and reproducibility of the biosensors were evaluated by applying them directly to beer real samples. It has been demonstrated that the Nafion-Lac/Sonogel-Carbon system is the more stable with a relative standard deviation of 3.3% (n=10), maintaining 84% of its stable response for at least three weeks. Estimation of polyphenol index in eight lager beers and a comparison of the results with those obtained by a classical method was carried out.

Dual laccase-tyrosinase based Sonogel-Carbon biosensor for monitoring polyphenols in beers.

A biosensor based on the bi-immobilization of laccase and tyrosinase phenoloxidase enzymes has been successfully developed. This biosensor employs as the electrochemical transducer the Sonogel-Carbon, a novel type of electrode developed by our group. The immobilization step was accomplished by doping the electrode surface with a mixture of the enzymes, glutaricdialdehyde, and Nafion-ion exchanger, as protective additive. The response of this biosensor, denoted the dual Trametes versicolor laccase (La) and Mushroom tyrosinase (Ty) based Sonogel-Carbon, was optimized directly in beer real samples and its analytical performance with respect to five individual polyphenols was evaluated. The Lac-Ty/Sonogel-Carbon electrode responds to nanomolar concentrations of flavan-3-ols, hydroxycinnamic acids, and hydroxybenzoic acids. The limit of detection, sensitivity, and linear range for caffeic acid, taken as an example, were 26 nM, 167.53 nA M (-1), and 0.01-2 microM, respectively. In addition, the stability and reproducibility of the biosensor were also evaluated in beer samples. The Lac-Ty/sonogel-carbon electrode was verified as very stable in this matrix, maintaining 80% of its stable response for at least three weeks, with a RSD of 3.6% ( n = 10). The biosensor was applied to estimate the total polyphenol index in ten beer samples, and a correlation of 0.99 was obtained when the results were compared with those obtained using the Folin-Ciocalteau reagent.

The Sonogel-Carbon materials as basis for development of enzyme biosensors for phenols and polyphenols monitoring: a detailed comparative study of three immobilization matrixes.

Three amperometric biosensors based on immobilization of tyrosinase on a new Sonogel-Carbon electrode for detection of phenols and polyphenols are described. The electrode was prepared using high energy ultrasounds (HEU) directly applied to the precursors. The first biosensor was obtained by simple adsorption of the enzyme on the Sonogel-Carbon electrode. The second and the third ones, presenting sandwich configurations, were initially prepared by adsorption of the enzyme and then modification by mean of polymeric membrane such as polyethylene glycol for the second one, and the ion-exchanger Nafion in the case of the third biosensor. The optimal enzyme loading and polymer concentration, in the second layer, were found to be 285 U and 0.5%, respectively. All biosensors showed optimal activity at the following conditions: pH 7, -200 mV, and 0.02 mol l(-1) phosphate buffer. The response of the biosensors toward five simple phenols derivatives and two polyphenols were investigated. It was found that the three developed tyrosinase Sonogel-Carbon based biosensors are in satisfactory competitiveness for phenolic compounds determination with other tyrosinase based biosensors reported in the literature. The detection limit, sensitivity, and the apparent Michaelis-Menten constant K(m)(app) for the Nafion modified biosensor were, respectively, 0.064, 0.096, and 0.03 micromol, 82.5, 63.4, and 194 nA micromol(-1)l(-1), and 67.1, 54.6, and 12.1 micromol l(-1) for catechol, phenol, and 4-chloro-3-methylphenol. Hill coefficient values (around 1 for all cases), demonstrated that the immobilization method does not affect the nature of the enzyme and confirms the biocompatibility of the Sonogel-Carbon with the bioprobe. An exploratory application to real samples such as beers, river waters and tannery wastewaters showed the ability of the developed Nafion/tyrosinase/Sonogel-Carbon biosensor to retain its stable and reproducible response.

Innovación en el concepto de peeling:CIMEL ICP, con acción despigmentante y rejuvenecedora / Innovation in peelings: cimel, with despigmenting and rejuvenating action

En el campo de la Dermatología Cosmética contamos con múltiples armas terapéuticas. Los peelings representan una de las más utilizadas, especialmente los superficiales y medios, en las consultas del dermatólogo. Se describe un nuevo concepto de peeling semioclusivo denominado Cimel, orientado hacia el tratamiento despigmentante, rejuvenecedor y antiseborreico de la piel. Material y métodos: El peeling Cimel se compone de la combinación de una serie de sustancias a altas concentraciones que se aplica en forma de mascarilla durante un tiempo de 4 a 8 horas. A su composición base se añaden diversos principios activos en función de la lesión predominante a tratar. Para el mantenimiento se utilizó el Cimel Conservador-C. Han sido tratados 183 casos, 60 con hiperpigmentaciones de diversa índole, 55 con signos de envejecimiento cutáneo, 42 con piel seborreica y 26 con pigmentaciones melano-hemáticas. La valoración de los resultados se efectuó mediante inspección visual por parte de los investigadores y mediante fotografías antes y después del procedimiento. En 3 casos se procedió a la realización de biopsia cutánea antes y después del peeling. Resultados: En el grupo de las hiperpigmentaciones, se observaron resultados excelentes en un 90 por ciento de los casos, en el envejecimiento en un 80 por ciento, en la piel seborreica en un 75 por ciento y en las pigmentaciones melano-hemáticas en un 70 por ciento. Los efectos inmediatos fueron eritema y fina descamación. Las complicaciones fueron nulas. Conclusión: El peeling Cimel supone una gran innovación en el campo de la Dermocosmética por sus resultados y su polivalencia, al abarcar diversas indicaciones (AU)