Application of molecular biological methods for studying probiotics and the gut flora

Increasingly, the microbiological scientific community is relying on molecular biology to define the complexity of the gut flora and to distinguish one organism from the next. This is particularly pertinent in the field of probiotics, and probiotic therapy, where identifying probiotics from the commensal flora is often warranted. Current techniques, including genetic fingerprinting, gene sequencing, oligonucleotide probes and specific primer selection, discriminate closely related bacteria with varying degrees of success. Additional molecular methods being employed to determine the constituents of complex microbiota in this area of research are community analysis, denaturing gradient gel electrophoresis (DGGE)/temperature gradient gel electrophoresis (TGGE), fluorescent in situ hybridisation (FISH) and probe grids. Certain approaches enable specific aetiological agents to be monitored, whereas others allow the effects of dietary intervention on bacterial populations to be studied. Other approaches demonstrate diversity, but may not always enable quantification of the population. At the heart of current molecular methods is sequence information gathered from culturable organisms. However, the diversity and novelty identified when applying these methods to the gut microflora demonstrates how little is known about this ecosystem. Of greater concern is the inherent bias associated with some molecular methods. As we understand more of the complexity and dynamics of this diverse microbiota we will be in a position to develop more robust molecular-based technologies to examine it. In addition to identification of the microbiota and discrimination of probiotic strains from commensal organisms, the future of molecular biology in the field of probiotics and the gut flora will, no doubt, stretch to investigations of functionality and activity of the microflora, and/or specific fractions. The quest will be to demonstrate the roles of probiotic strains in vivo and not simply their presence or absence.

Bibliographic analysis of persuasive systems: techniques, methods and domains of application

This paper presents findings of our study on peer-reviewed papers published in the International Conference on Persuasive Technology from 2006 to 2010. The study indicated that out of 44 systems reviewed, 23 were reported to be successful, 2 to be unsuccessful and 19 did not specify whether or not it was successful. 56 different techniques were mentioned and it was observed that most designers use ad hoc definitions for techniques or methods used in design. Hence we propose the need for research to establish unambiguous definitions of techniques and methods in the field.

Methods of Estimation in Multiple Linear Regression: Application to Clinical Data

Nesse artigo, tem-se o interesse em avaliar diferentes estratégias de estimação de parâmetros para um modelo de regressão linear múltipla. Para a estimação dos parâmetros do modelo foram utilizados dados de um ensaio clínico em que o interesse foi verificar se o ensaio mecânico da propriedade de força máxima (EM-FM) está associada com a massa femoral, com o diâmetro femoral e com o grupo experimental de ratas ovariectomizadas da raça Rattus norvegicus albinus, variedade Wistar. Para a estimação dos parâmetros do modelo serão comparadas três metodologias: a metodologia clássica, baseada no método dos mínimos quadrados; a metodologia Bayesiana, baseada no teorema de Bayes; e o método Bootstrap, baseado em processos de reamostragem.

Computational aspects of harmonic wavelet Galerkin methods and an application to a precipitation front propagation model

This article is dedicated to harmonic wavelet Galerkin methods for the solution of partial differential equations. Several variants of the method are proposed and analyzed, using the Burgers equation as a test model. The computational complexity can be reduced when the localization properties of the wavelets and restricted interactions between different scales are exploited. The resulting variants of the method have computational complexities ranging from O(N(3)) to O(N) (N being the space dimension) per time step. A pseudo-spectral wavelet scheme is also described and compared to the methods based on connection coefficients. The harmonic wavelet Galerkin scheme is applied to a nonlinear model for the propagation of precipitation fronts, with the front locations being exposed in the sizes of the localized wavelet coefficients. (C) 2011 Elsevier Ltd. All rights reserved.

Expedient, accurate methods for the determination of the degree of substitution of cellulose carboxylic esters: Application of UV-vis spectroscopy (dye solvatochromism) and FTIR

Two techniques, namely UV-vis- and FTIR spectroscopy, have been employed in order to calculate the degree of substitution (DS) of cellulose carboxylic esters, including acetates, CAs, butyrates, CBs, and hexanoates, CHs. Regarding UV-vis spectroscopy, we have employed a novel approach, based on measuring the dependence of lambda(max) of the intra-molecular charge-transfer bands of polarity probes adsorbed on DS of the ester films (solvatochromism). Additionally, we have revisited the use of FTIR for DS determination. Several methods have been used in order to plot Beer`s law graph, namely: Absorption of KBr pellets, pre-coated with CA: reflectance (DRIFTS) of CAs-KBr solid-solid mixtures with, or without the use of 1.4-dicyanobenzene as an internal reference; reflectance of KBr powder pre-coated with CA. The methods indicated are simple, fast, and accurate, requiring much less ester than the titration method. The probe method is independent of the experimental variables examined. (c) 2010 Published by Elsevier Ltd.

Application of chemometric methods in the evaluation of chemical and spectroscopic data on organic matter from Oxisols in sewage sludge applications

Chemometric methods can contribute to soil research by permitting the extraction of more information from the data. The aim of this work was to use Principal Component Analysis to evaluate data obtained through chemical and spectroscopic methods on the changes in the humification process of soil organic matter from two tropical soils after sewage sludge application. In this case, humic acids extracted from Typic Eutrorthox and Typic Haplorthox soils with and without sewage sludge application for 7 consecutive years were studied. The results obtained for all of the samples and methods showed two clusters: samples extracted from the two soil types. These expected results indicated the textural difference between the two soils was more significant than the differences between treatments (control and sewage sludge application) or between depths. In this case, an individual chemometric treatment was made for each type of soil. It was noted that the characterization of the humic acids extracted from soils with and without sewage sludge application after 7 consecutive years using several methods supplies important results about changes in the humification degree of soil organic matter, These important result obtained by Principal Component Analysis justify further research using these methods to characterize the changes in the humic acids extracted from sewage sludge-amended soils. (C) 2009 Elsevier B.V. All rights reserved.

Application of chemometric methods in the evaluation of chemical and spectroscopic data on organic matter from Oxisols in sewage sludge applications

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

Development and application of methods for determination of residual monomer in dental acrylic resins using high performance liquid chromatography

Two high-performance liquid chromatographic methods for determination of residual monomer in dental acrylic resins are described. Monomers were detected by their UV absorbance at 230 nm, on a Nucleosil((R)) C-18 (5 mu m particle size, 100 angstrom pore size, 15 x 0.46 cm i.d.) column. The separation was performed using acetonitrile-water (55:45 v/v) containing 0.01% triethylamine (TEA) for methyl methacrylate and butyl methacrylate, and acetonitrile-water (60:40 v/v) containing 0.01% TEA for isobutyl methacrylate and 1,6-hexanediol dimethacrylate as mobile phases, at a flow rate of 0.8 mL/min. Good linear relationships were obtained in the concentration range 5.0-80.0 mu g/mL for methyl methacrylate, 10.0-160.0 mu g/mL for butyl methacrylate, 50.0-500.0 mu g/mL for isobutyl methacrylate and 2.5-180.0 mu g/mL for 1,6-hexanediol dimethacrylate. Adequate assay for intra- and inter-day precision and accuracy was observed during the validation process. An extraction procedure to remove residual monomer from the acrylic resins was also established. Residual monomer was obtained from broken specimens of acrylic disks using methanol as extraction solvent for 2 h in an ice-bath. The developed methods and the extraction procedure were applied to dental acrylic resins, tested with or without post-polymerization treatments, and proved to be accurate and precise for the determination of residual monomer content of the materials evaluated. Copyright (c) 2005 John Wiley & Sons, Ltd.

Influence of Methods of Application of Self-etching Adhesive Systems on Adhesive Bond Strength to Enamel

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

Influence of two methods of CaCl2 application and concentrations used to maintain passion fruit firmness

This is a report on the influence of the methods of postharvest application of CaCl2 in several concentrations, on the maintenance of the firmness of the sweet passion fruit (Passiflora alata Dryander). The fruits, picked in the pre-climateric stage, were selected desinfection with thiabendazol and submitted to CaCl2 in concentrations of 0% (control), 1%, 2%, 3% or 4% with two methods: immersion (for 2 h) and infiltration under vacuum and pressure (- 25 vacuum kPa, for 1 minute and, + 25 pressure kPa, for 1 min). All fruits were stored under refrigeration (9 degreesC, 85-90% RH), for 30 d. The fruits were evaluated for texture every 3 d. From the results, it is concluded that application of CaCl2 by infiltration favored the maintenance of fruit consistency during the first 18 days of storage. Fruits treated with 1% and 2% of CaCl2 were firmer than the others.

Effect of different methods of cutting fluid application on turning of a difficult-to-machine steel (SAE EV-8)

In this study, different methods of cutting fluid application are used in turning of a difficult-to-machine steel (SAE EV-8). Initially, a semisynthetic cutting fluid was applied using a conventional method (i.e. overhead flood cooling), minimum quantity of cutting fluid, and pulverization. A lubricant of vegetable oil (minimum quantity of lubricant) was also applied using the minimum quantity method. Thereafter, a cutting fluid jet under high pressure (3.0 MPa) was singly applied in the following regions: chip-tool interface, top surface of the chip (between workpiece and chip) and tool-workpiece contact. Moreover, two other methods were used: an interflow between conventional application and chip-tool interface jet (combined method) and, finally, three jets simultaneously applied. In order to carry out these tests, it was necessary to set up a high-pressure system using a piston pump for generating a cutting fluid jet, a venturi for fluid application (minimum quantity of cutting fluid and minimum quantity of lubricant) and a nozzle for cutting fluid pulverization. The output variables analyzed included tool life, surface roughness, cutting tool temperature, cutting force, chip form, chip compression rate and machined specimen microstructure. Among the results, it can be observed that the tool life increases and the cutting force decreases with the application of cutting fluid jet, mainly when it is directed to the chip-tool interface. Excluding the methods involving jet fluid, the conventional method seems to be more efficient than other methods of low pressure, such as minimum quantity of volume and pulverization, when considering just the cutting tool wear. © 2013 IMechE.

Different methods of cutting fluid application on turning of a difficult-to-machine steel

Different methods of cutting fluid application are used on turning of a difficult-tomachine steel (SAE EV-8). A semi-synthetic cutting fluid was applied using a conventional method, minimum quantity of cutting fluid (MQCF), and pulverization. By the minimum quantity method was also applied a lubricant of vegetable oil (MQL). Thereafter, a cutting fluid jet under high pressure (3.0 MPa) was singly applied in the following regions: chip-tool interface; top surface of the chip; and tool-workpiece contact. Two other methods were used: an interflow between conventional application and chip-tool interface jet and, finally, three jets simultaneously applied. In order to carry out these tests, it was necessary to set up a high pressure system using a piston pump for generating a cutting fluid jet, a Venturi for fluid application (MQCF and MQL), and a nozzle for cutting fluid pulverization. The output variables analyzed included tool life, surface roughness, cutting tool temperature, cutting force, chip form, chip compression rate and machined specimen microstructure. It can be observed that the tool life increases and the cutting force decreases with the application of cutting fluid jet, mainly when it is directed to the chip-tool interface. Excluding the methods involving jet fluid, the conventional method seems to be more efficient than other methods of low pressure. © (2013) Trans Tech Publications, Switzerland.

Estimating potential output: a survey of the alternative methods and their application to Brazil

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