Elution of proteins from polyacrylamide eels: a simple and economic procedure

A simplified methodology for the quantitative electroelution of proteins from polyacrylamide gels is described. After staining with Coomassie Brilliant Blue R 250, the identified bands are excised from the gel and the proteins eluted using a procedure developed for use in conventional tube gel electrophoresis equipment.

A stratigraphic framework for the Miocene from the lower Tagus Basin (Lisbon, Setúbal Península, Portugal) Depositional sequences, biostratigraphy and isotopic ages

Rev. Soc. Geol. España, 12(1), ano 1999

In-plane displacement and strain image analysis

Measurements in civil engineering load tests usually require considerable time and complex procedures. Therefore, measurements are usually constrained by the number of sensors resulting in a restricted monitored area. Image processing analysis is an alternative way that enables the measurement of the complete area of interest with a simple and effective setup. In this article photo sequences taken during load displacement tests were captured by a digital camera and processed with image correlation algorithms. Three different image processing algorithms were used with real images taken from tests using specimens of PVC and Plexiglas. The data obtained from the image processing algorithms were also compared with the data from physical sensors. A complete displacement and strain map were obtained. Results show that the accuracy of the measurements obtained by photogrammetry is equivalent to that from the physical sensors but with much less equipment and fewer setup requirements. © 2015Computer-Aided Civil and Infrastructure Engineering.

Error control on spectral data of four-wave mixing based on a-SiC technology

In this paper we exploit the nonlinear property of the SiC multilayer devices to design an optical processor for error detection that enables reliable delivery of spectral data of four-wave mixing over unreliable communication channels. The SiC optical processor is realized by using double pin/pin a-SiC:H photodetector with front and back biased optical gating elements. Visible pulsed signals are transmitted together at different bit sequences. The combined optical signal is analyzed. Data show that the background acts as selector that picks one or more states by splitting portions of the input multi optical signals across the front and back photodiodes. Boolean operations such as EXOR and three bit addition are demonstrated optically, showing that when one or all of the inputs are present, the system will behave as an XOR gate representing the SUM. When two or three inputs are on, the system acts as AND gate indicating the present of the CARRY bit. Additional parity logic operations are performed using four incoming pulsed communication channels that are transmitted and checked for errors together. As a simple example of this approach, we describe an all-optical processor for error detection and then provide an experimental demonstration of this idea. (C) 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Simple laccase-based biosensor for formetanate hydrochloride quantification in fruits

This work describes the development of an electrochemical enzymatic biosensor for quantification of the pesticide formetanate hydrochloride (FMT). It is based on a gold electrode modified with electrodeposited gold nanoparticles and laccase. The principle behind its development relies on FMT's capacity to inhibit the laccase catalytic reaction that occurs in the presence of phenolic substrates. The optimum values for the relevant experimental variables such as gold nanoparticles electrochemical deposition (at − 0.2 V for 100 s), laccase immobilization (via glutaraldehyde cross-linking), laccase concentration (12.4 mg/mL), substrate selection and concentration (5.83×10−5 M of aminophenol), pH (5.0), buffer (Britton–Robinson), and square-wave voltammetric parameters were determined. The developed biosensor was successfully applied to FMT determination in mango and grapes. The attained limit of detection was 9.5×10−8 ± 9.5×10−10 M (0.02 ± 2.6×10−4 mg/kg on a fresh fruit weight basis). Recoveries for the five tested spiking levels ranged from 95.5 ± 2.9 (grapes) to 108.6 ± 2.5% (mango). The results indicated that the proposed device presents suitable characteristics in terms of sensitivity (20.58 ± 0.49 A/μM), linearity (9.43×10−7 to 1.13×10−5 M), accuracy, repeatability (RSD of 1.4%), reproducibility (RSD of 1.8%) and stability (19 days) for testing of compliance with established maximum residue limits of FMT in fruits and vegetables.

Simple and stable dynamic traffic engineering for provider scale ethernet

Trabalho apresentado no âmbito do Mestrado em Engenharia Informática, como requisito parcial para obtenção do grau de Mestre em Engenharia Informática

Enzyme linked immunosorbent assay for rubella antibodies: a simple method of antigen production. A preliminary report

A simple method of rubella antigen production by treatment with sodium desoxycholate for use in enzyme immunoassay (IMT-ELISA) is presented. When this assay was compared with a commercial test (Enzygnost-Rubella, Behring), in the study of 108 sera and 118 filter paper blood samples, 96.9% (219/226) overall agreement and correlation coefficient of 0.90 between absorbances were observed. Seven samples showed discordant results, negative by the commercial kit and positive by our test. Four of those 7 samples were available, being 3 positive by HI.

A simple and inexprensive method to generate a microaerophilic atmosphere for the isolation of Campylobacter sp

Faeces of 138 chickens were inoculated on Blaser agar plates. One set of plates was incubated in jars with CampyPak envelopes. The others were incubated in "Zip-lock" plastic bags (7 x X in.) and a microaerophilic atmosphere was generated exhaling into the "Zip-lock" plastic bag, after holding the breath for 20 sec. Then, the bag was pressed to evacuate its atmosphere, inflated again, and pressed (4 times), and finally sealed. Campylobacter was isolated from 127 (96.2%) of samples incubated in jars with gas generator envelopes and from 129 (98%) of the specimens incubated into the bags. The proposed methodology offers good savings for cost-conscious laboratories.

A simple PCR-based procedure for plaque diagnosis

Supernatant of boiled spleen saline-suspensions of Yersinia pestis experimentally infected animals were used as template for PCR amplification without DNA extraction. PCR sensitivity was enhanced by a second round of amplification (Nested). No amplification was observed from non-infected animals.

The use of oligonucleotide probes for meningococcal serotype characterization

In the present study we examine the potential use of oligonucleotide probes to characterize Neisseria meningitidis serotypes without the use of monoclonal antibodies (MAbs). Antigenic diversity on PorB protein forms the bases of serotyping method. However, the current panel of MAbs underestimated, by at least 50% the PorB variability, presumably because reagents for several PorB variable regions (VRs) are lacking, or because a number of VR variants are not recognized by serotype-defining MAbs12. We analyzed the use of oligonucleotide probes to characterize serotype 10 and serotype 19 of N. meningitidis. The porB gene sequence for the prototype strain of serotype 10 was determined, aligned with 7 other porB sequences from different serotypes, and analysis of individual VRs were performed. The results of DNA probes 21U (VR1-A) and 615U (VR3-B) used against 72 N. meningitidis strains confirm that VR1 type A and VR3 type B encode epitopes for serotype-defined MAbs 19 and 10, respectively. The use of probes for characterizing serotypes possible can type 100% of the PorB VR diversity. It is a simple and rapid method specially useful for analysis of large number of samples.

Rapid Evolution of the Sequences and Gene Repertoires

Proteins secreted to the extracellular environment or to the periphery of the cell envelope, the secretome, play essential roles in foraging, antagonistic and mutualistic interactions. We hypothesize that arms races, genetic conflicts and varying selective pressures should lead to the rapid change of sequences and gene repertoires of the secretome. The analysis of 42 bacterial pan-genomes shows that secreted, and especially extracellular proteins, are predominantly encoded in the accessory genome, i.e. among genes not ubiquitous within the clade. Genes encoding outer membrane proteins might engage more frequently in intra-chromosomal gene conversion because they are more often in multi-genic families. The gene sequences encoding the secretome evolve faster than the rest of the genome and in particular at non-synonymous positions. Cell wall proteins in Firmicutes evolve particularly fast when compared with outer membrane proteins of Proteobacteria. Virulence factors are over-represented in the secretome, notably in outer membrane proteins, but cell localization explains more of the variance in substitution rates and gene repertoires than sequence homology to known virulence factors. Accordingly, the repertoires and sequences of the genes encoding the secretome change fast in the clades of obligatory and facultative pathogens and also in the clades of mutualists and free-living bacteria. Our study shows that cell localization shapes genome evolution. In agreement with our hypothesis, the repertoires and the sequences of genes encoding secreted proteins evolve fast. The particularly rapid change of extracellular proteins suggests that these public goods are key players in bacterial adaptation.

FATAL GROUP A STREPTOCOCCAL TOXIC SHOCK-LIKE SYNDROME IN A CHILD WITH VARICELLA: REPORT OF THE FIRST WELL DOCUMENTED CASE WITH DETECTION OF THE GENETIC SEQUENCES THAT CODE FOR EXOTOXINS SPE A AND B, IN SÃO PAULO, BRAZIL

A previously healthy seven-year-old boy was admitted to the intensive care unit because of toxaemia associated with varicella. He rapidly developed shock and multisystem organ failure associated with the appearance of a deep-seated soft tissue infection and, despite aggressive treatment, died on hospital day 4. An M-non-typable, spe A and spe B positive Group A Streptococcus was cultured from a deep soft tissue aspirate. The criteria for defining Streptococcal toxic shock-like syndrome were fulfilled. The authors discuss the clinical and pathophysiological aspects of this disease as well as some unusual clinical findings related to this case.

Estudo de delaminação em compósitos de matriz polimérica

A aplicação do material compósito é neste momento bastante vasta, graças à combinação das suas características específicas, tais como, maior resistência específica e módulos específicos e melhor resistência à fadiga, quando comparados com os metais convencionais. Tais características, quando requeridas, tornam este material ideal para aplicações estruturais. Esta caminhada de sucesso iniciou desde muito cedo, quando o material compósito já era utilizado para fabrico de armas pelos mongóis e na construção civil pelos hebreus e egípcios, contudo, só a partir dos meados do século XX é que despertou interesses para aplicações mais modernas. Atualmente os materiais compósitos são utilizados em equipamentos domésticos, componentes elétricos e eletrónicos, passando por materiais desportivos, pela indústria automóvel e construção civil, até indústrias de grande exigência e visibilidade tecnológica como a aeronáutica, espacial e de defesa. Apesar das boas características apresentadas pelos materiais compósitos, no entanto, estes materiais têm tendência a perderem as suas propriedades quando submetidas a algumas operações de acabamento como a furação. A furação surge da necessidade de ligação de peças de um mesmo mecanismo. Os furos obtidos por este processo devem ser precisos e sem danos para garantir ligações de alta resistência e também precisas. A furação nos materiais compósitos é bastante complexa devido à sua heterogeneidade, anisotropia, sensibilidade ao calor e pelo facto de os reforços serem extremamente abrasivos. A operação de furação pode causar grandes danos na peça, como a delaminação a entrada, defeitos de circularidade do furo, danos de origem térmica e a delaminação à saída que se apresenta como o mais frequente e indesejável. Com base nesses pressupostos é que este trabalho foi desenvolvido de forma a tentar obter processos simples para determinação e previsão de danos em polímeros reforçados com fibras (de carbono neste caso) de forma a precavê-los. De forma a conseguir estes objetivos, foram realizados ensaios de início de delaminação segundo a proposta de Lachaud et al. e ensaios de pin-bearing segundo a proposta de Khashaba et al. Foram também examinadas extensões de danos de acordo com o modelo de Fator de delaminação ajustado apresentado por Davim et al. A partir dos ensaios, de pin-bearing, realizados foram analisadas influências do material e geometria da broca, do avanço utilizado na furação e de diferentes orientações de empilhamentos de placas na delaminação de laminados compósitos e ainda a influências dessas variáveis na força de rutura por pin-bearing. As principais conclusões tiradas daqui são que a delaminação aumenta com o aumento do avanço, o que já era esperado, as brocas em carboneto de tungsténio são as mais recomendas para a furação do material em causa e que a delaminação é superior para a placa cross-ply quando comparada com placas unidirecionais. Para a situação de ensaios de início de delaminação foram analisadas as influências da variação da espessura não cortada por baixo da broca/punção, de diferentes geometrias de brocas, da alteração de velocidade de ensaio e diferentes orientações de empilhamentos de placas na força de início de delaminação. Deste ensaio as principais conclusões são que a força de início de delaminação aumenta com o aumenta da espessura não cortada e a influência da velocidade de ensaio altera com a variação das orientações de empilhamento.

Une langue étragère, un ordinateur, une image: c'est simple comme bonjour!

Este artigo surgiu na sequência de um atelier “Une langue étrangère, un ordinateur, une image: c’est simple comme bonjour!”, desenvolvido no âmbito do XXI Congresso da Associação Portuguesa dos Professores de Francês, Images et imaginaires pour agir. Teve como propósito divulgar, experimentar e refletir sobre recursos digitais que podem dar um bom contributo ao processo de ensino e aprendizagem do Francês Língua Estrangeira (FLE). Evidencia-se o poder da imagem na construção do conhecimento, desafiando a criatividade e novos modos de ensinar a aprender. Verificou-se que os professores se interessaram pelas ferramentas digitais e evidenciaram a sua importância e a sua aplicabilidade nos contextos educativos. Neste sentido, o artigo divulga ferramentas informáticas focadas no desenvolvimento da oralidade/leitura/escrita do francês língua estrangeira, refere boas práticas de utilização em contexto de sala de aula, constituindo uma contribuição para a renovação da escola.