Reagentless biosensor for isocitrate using one step modified Pt-Ir microelectrode

The Pt-Ir microelectrode modified through one step, electropolymerization is proposed for the isocitrate amperometric biosensor construction. The enzyme (isocitrate dehydrogenase-ICDH), coenzyme (NADP(+)) and mediator (Meldola's Blue) were immobilized onto the microelectrode surface in one step from a PIPES buffer solution containing pyrrole. The optimized experimental conditions were 25 cycles of cyclic voltammetric in a solution containing 3.58 10(-5) mol l(-1) of mediator, 3.51 10(-4) mol l(-1) of coenzyme and 2.68 U ml(-1) of enzyme. In contrast to the biosensor for isocitrate reported in literature, just one enzyme was immobilized and no coenzyme addition in the solution of analysis was necessary. Catalytic currents were proportional to the isocitrate concentration between 7.7 10(-6) and 1.04 10(-4) mol l(-1), showing good repeatability. The detection limit of the proposed biosensor was 3.50 10(-6) mol l(-1), the response time was lower than 20 s, the lifetime was about 30 determinations and no significant interference of sugars and citric acid was verified. Orange juice samples were analysed by both methodology biosensor and spectrophotometric commercial kit, and the obtained results presented a good correlation. The data demonstrated that the developed biosensor is suitable for isocitrate determination in orange juice without matrix interferences. (C) 2001 Elsevier B.V. B.V. All rights reserved.

One-step purification of 5-enolpyruvylshikimate-3-phosphate synthase enzyme from Mycobacterium tuberculosis

Currently, there are 8 million new cases and 2 million deaths annually from tuberculosis, and it is expected that a total of 225 million new cases and 79 million deaths will occur between 1998 and 2030. The reemergence of tuberculosis as a public health threat, the high susceptibility of HIV-infected persons, and the proliferation of multi-drug-resistant strains have created a need to develop new antimycobacterial agents. The existence of homologues to the shikimate pathway enzymes has been predicted by the determination of the genome sequence of Mycobacterium tuberculosis. We have previously reported the cloning and overexpression of M. tuberculosis aro A-encoded EPSP synthase in both soluble and active forms, without IPTG induction. Here, we describe the purification of M. tuberculosis EPSP synthase (mtEPSPS) expressed in Escherichia coli BL21(DE3) host cells. Purification of mtEPSPS was achieved by a one-step purification protocol using an anion exchange column. The activity of the homogeneous enzyme was measured by a coupled assay using purified shikimate kinase and purine nucleoside phosphorylase proteins. A total of 53 mg of homogeneous enzyme could be obtained from 1 L of LB cell culture, with a specific activity value of approximately 18 U mg-1. The results presented here provide protein in quantities necessary for structural and kinetic studies, which are currently underway in our laboratory. © 2002 Elsevier Science (USA). All rights reserved.

Evaluation of a two-step pouring technique for implant-supported prostheses impression

The aim of this study was to evaluate the efficacy of a pouring technique for implant-supported prostheses impressions. A metallic matrix (control group) with two implants positioned at 90 and 65 degrees was fabricated. The matrix was submitted to the direct transfer impression technique. In group CP (conventional pouring - n = 10), casts were obtained by the conventional pouring technique. In group EP (experimental pouring - n = 10), the analogs were embraced with latex tubes before the first pouring and then submitted to a second pouring. Vertical misfit and implants/analogs inclinations were evaluated. Data were analyzed by analysis of variance and Tukey's test (p < .05). Results demonstrated significant difference (p < .05) between control and experimental groups for misfit measurement in perpendicular implant/analog and between control group and group EP in leaning implant/analog. Considering inclination, there were significant differences (p < .05) between control and experimental groups for leaning analogs. Independently of the pouring technique, perpendicular implants produced more accurate casts.

A DC-DC converter based on the three-state switching cell for high current and voltage step-down applications

This paper presents a pulsewidth modulation dc-dc nonisolated buck converter using the three-state switching cell, constituted by two active switches, two diodes, and two coupled inductors. Only part of the load power is processed by the active switches, reducing the peak current through the switches to half of the load current, as higher power levels can then be achieved by the proposed topology. The volume of reactive elements, i.e., inductors and capacitors, is also decreased since the ripple frequency of the output voltage is twice the switching frequency. Due to the intrinsic characteristics of the topology, total losses are distributed among all semiconductors. Another advantage of this converter is the reduced region for discontinuous conduction mode when compared to the conventional buck converter or, in other words, the operation range in continuous conduction mode is increased, as demonstrated by the static gain plot. The theoretical approach is detailed through qualitative and quantitative analyses by the application of the three-state switching cell to the buck converter operating in nonoverlapping mode $(D < 0.5)$. Besides, the mathematical analysis and development of an experimental prototype rated at 1 kW are carried out. The main experimental results are presented and adequately discussed to clearly identify its claimed advantages. © 1986-2012 IEEE.

Comparison between the six-minute walk test and the six-minute step test in post stroke patients

Background: The Stroke remains one of the major chronic diseases worldwide, and is considered a major cause of disability, which results not only in persistent neurological deficits, but also in the high physical deconditioning, nevertheless there are not many forms of assessing functional capacity in this population. We aimed to investigate the feasibility of the Six Minute Walk Teste and the Six-Minute Step Test (6MST) in post-stroke patients and compare the behavior of physiological variables during the 6MST and the Six-Minute Walk Test (6MWT), by correlating the functional performance obtained in both tests. Method. The 6MWT was carried out according to the American Thoracic Society (ATS) and the 6MST was performed in six minutes in order to compare it to the 6MWT in a 20 cm step. Was included post-stroke individuals able to walk without aid. All of them did the 6MWT and the 6MST. Results: 12 patients participated in the study. There was no statistical difference in the parameters analyzed when tests were compared. There was poor correlation between the functional performance in both tests. Conclusion: The 6MWT and the 6MST is feasible for post-stroke patients and physiological responses are equal during the performance of both tests. However, there was no correlation with respect to functional performance, which was assessed by the distance walked in the 6MWT and by the number of steps climbed in the 6MST. © 2013 da Silva et al.; licensee BioMed Central Ltd.

LED light attenuation through human dentin: A first step toward pulp photobiomodulation after cavity preparation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

One step forward: contrasting the effects of Toe clipping and PIT tagging on frog survival and recapture probability

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Phase space properties and chaotic transport for a particle moving in a time dependent step potential well

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Performance of a New One-step Multi-mode Adhesive on Etched vs Non-etched Enamel on Bond Strength and Interfacial Morphology

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of nicotine and cotinine impregnation on the first step of periodontal regeneration: clot stabilization

This study analyzes the clot stabilization on root surfaces of teeth impregnated with cotinine and nicotine and the influence of the scaling in the adhesion of blood components, observing the influence of new exposition to nicotine and/or cotinine after scaling. Fifteen human teeth extracted due to periodontal disease of non-smokers patients were selected and manually scaled. Four dentin blocks were obtained from each tooth (n = 60). Samples received blood application or reimpregnation with nicotine and/or cotinine, depending on the groups. Group 1: PBS immersion + root scaling + blood; group 2: nicotine + root scaling + blood; group 3: nicotine + root scaling + nicotine reapplication + blood; group 4: cotinine + root scaling + blood; group 5: cotinine + root scaling + cotinine reapplication+ blood; group 6: nicotine and cotinine + root scaling + nicotine and cotinine + blood. Samples were kept in 2 ml of each substance for 24 hours. Each group received a blood drop and was analyzed by SEM. The higher amount of blood components was present in teeth exposed to cotinine and the groups submitted to scaling and blood application in comparison with groups that received reapplication of toxic substances after scaling. The greater toxic effect on root dentin surface was after the exposure to nicotine and cotinine. Results suggest that periodontal healing may be delayed in smokers due to the direct inhibition of clot stabilization on the root surface when nicotine and cotinine are present concomitantly.

A step to the gigantic genome of the desert locust: chromosome sizes and repeated DNAs

The desert locust (Schistocerca gregaria) has been used as material for numerous cytogenetic studies. Its genome size is estimated to be 8.55 Gb of DNA comprised in 11 autosomes and the X chromosome. Its X0/XX sex chromosome determinism therefore results in females having 24 chromosomes whereas males have 23. Surprisingly, little is known about the DNA content of this locust's huge chromosomes. Here, we use the Feulgen Image Analysis Densitometry and C-banding techniques to respectively estimate the DNA quantity and heterochromatin content of each chromosome. We also identify three satellite DNAs using both restriction endonucleases and next-generation sequencing. We then use fluorescent in situ hybridization to determine the chromosomal location of these satellite DNAs as well as that of six tandem repeat DNA gene families. The combination of the results obtained in this work allows distinguishing between the different chromosomes not only by size, but also by the kind of repetitive DNAs that they contain. The recent publication of the draft genome of the migratory locust (Locusta migratoria), the largest animal genome hitherto sequenced, invites for sequencing even larger genomes. S. gregaria is a pest that causes high economic losses. It is thus among the primary candidates for genome sequencing. But this species genome is about 50 % larger than that of L. migratoria, and although next-generation sequencing currently allows sequencing large genomes, sequencing it would mean a greater challenge. The chromosome sizes and markers provided here should not only help planning the sequencing project and guide the assembly but would also facilitate assigning assembled linkage groups to actual chromosomes.

One-step glycerol oxidehydration to acrylic acid on multifunctional zeolite catalysts

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Resin tag length of one-step and self-etching adhesives bonded to unground enamel

Length of resin tags yielded by utilization of an one-step conventional adhesive system and self-etching adhesive system on unground enamel was observed. In study Groups I and III, the enamel surface was etched for 60 seconds with 35% phosphoric acid gel and adhesive systems PQ1 (Ultradent Products, Inc) and Adper Prompt L Pop (3M/ESPE) were applied. Adper Prompt L Pop (3M/ESPE) was also applied in Group II in accordance with the manufacturer's recommendations. After application of these adhesive systems to dental enamel, specimens were prepared for light microscopy analysis to ascertain degree of penetration (x400). The results were submitted to an analysis of variance at the 5% level; whenever there was significance, the Tukey test was applied at the 5% level. It was found that acid etching prior to application of conventional and self-etching adhesive materials provided higher penetration of the adhesive into the unground enamel surface compared to that achieved solely by application of self-etching adhesive.

Dextran sulfate/Triton X two-phase micellar systems as an alternative first purification step for clavulanic acid

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Deterministic Solutions for a Step-growth Polymerization

Chain topology, including branch node, chain link and cross-link dynamics that contribute to the number of elastically active strands and junctions, are calculated using purely deterministic derivations. Solutions are not coupled to population density distributions. An eigenzeit transformation assists in the conversion of expressions derived by chemical reaction principles from time to conversion space, yielding transport phenomena type expressions where the rate of change in the molar concentrations of branch nodes with respect to conversion is expressed as functions of the fraction of reactive sites on precursors and reactants. Analogies are hypothesized to exist in cross-linking space that effectively distribute branch nodes with i reacted moieties between cross-links having j bonds extending to the gel. To obtain solutions, reacted sites on nodes or links with finite chain extensions are examined in terms of stoichiometry associated with covalent bonding. Solutions replicate published results based on Miller and Macosko’s recursive procedure and results obtained from truncated weighted sums of population density distributions as suggested by Flory.