A general assay for antibody catalysis using acridone as a fluorescent tag.

A simple and highly sensitive catalysis assay is demonstrated based on analyzing reactions with acridonetagged compounds by thin-layer chromatography. As little as 1 pmol of product is readily visualized by its blue fluorescence under UV illumination and identified by its retention factor (Rf). Each assay requires only 10 microliters of solution. The method is reliable, inexpensive, versatile, and immediately applicable in repetitive format for screening catalytic antibody libraries. Three examples are presented: (i) the epoxidation of acridone labeled (S)-citronellol. The pair of stereoisomeric epoxides formed is resolved on the plate, which provides a direct selection method for enantioselective epoxidation catalysts. (ii) Oxidation of acridone-labeled 1-hexanol to 1-hexanal. The activity of horse liver alcohol dehydrogenase is detected. (iii) Indirect product labeling of released aldehyde groups by hydrazone formation with an acridone-labeled hydrazide. Activity of catalytic antibodies for hydrolysis of enol ethers is detected.

Quantitative measurement of intraorganelle pH in the endosomal-lysosomal pathway in neurons by using ratiometric imaging with pyranine.

Organelle acidification is an essential element of the endosomal-lysosomal pathway, but our understanding of the mechanisms underlying progression through this pathway has been hindered by the absence of adequate methods for quantifying intraorganelle pH. To address this problem in neurons, we developed a direct quantitative method for accurately determining the pH of endocytic organelles in live cells. In this report, we demonstrate that the ratiometric fluorescent pH indicator 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) is the most advantageous available probe for such pH measurements. To measure intraorganelle pH, cells were labeled by endocytic uptake of HPTS, the ratio of fluorescence emission intensities at excitation wavelengths of 450 nm and 405 nm (F450/405) was calculated for each organelle, and ratios were converted to pH values by using standard curves for F450/405 vs. pH. Proper calibration is critical for accurate measurement of pH values: standard curves generated in vitro yielded artifactually low organelle pH values. Calibration was unaffected by the use of culture medium buffered with various buffers or different cell types. By using this technique, we show that both acidic and neutral endocytically derived organelles exist in the axons of sympathetic neurons in different steady-state proportions than in the cell body. Furthermore, we demonstrate that these axonal organelles have a bimodal pH distribution, indicating a rapid acidification step in their maturation that reduces the average pH of a fraction of the organelles by 2 pH units while leaving few organelles of intermediate pH at steady state. Finally, we demonstrate a spatial gradient or organelle pH along axons, with the relative frequency of acidic organelles increasing with proximity to the cell body.

Análise teórica-experimental de perfis de aço formados a frio devido à instabilidade por distorção na flexão

Apresenta-se neste trabalho um estudo teórico-experimental sobre a instabilidade de perfis formados a frio submetidos à flexão. A instabilidade distorcional se faz comum na presença de tensões de compressão atuando sobre perfis enrijecidos e fabricados com aços de elevada resistência mecânica. A parte teórica abrange os métodos de cálculo analíticos e numéricos para a análise de instabilidade distorcional de perfis de seção aberta formados a frio. Na parte experimental inclui-se o estudo de perfis formados a frio com seções do tipo U enrijecidos submetidos aos ensaios à flexão. Nestes ensaios variou-se a altura de alma e espessura de chapa procurando-se abranger maior número de condições geométricas para análise da estabilidade distorcional. Inclui-se também a análise de instabilidade numérica dos perfis do programa experimental através do método de resistência direta via método das faixas finitas. Com base nos resultados experimentais, numéricos e na análise teórica do problema, verificou-se o procedimento adotado pela NBR14762/2001 e efetuou-se comparação entre curvas de resistência propostas para o dimensionamento de perfis formados a frio à flexão. Foi verificado que o fenômeno de instabilidade distorcional pode ser o estado limite último crítico para o dimensionamento dos perfis formados a frio.

Development and psychometric testing of the clinical learning environment, supervision and nurse teacher evaluation scale (CLES+T): The Spanish version

Background: The Clinical Learning Environment, Supervision and Nurse Teacher scale is a reliable and valid instrument to evaluate the quality of the clinical learning process in international nursing education contexts. Objectives: This paper reports the development and psychometric testing of the Spanish version of the Clinical Learning Environment, Supervision and Nurse Teacher scale. Design: Cross-sectional validation study of the scale. Setting: 10 public and private hospitals in the Alicante area, and the Faculty of Health Sciences (University of Alicante, Spain). Participants: 370 student nurses on clinical placement (January 2011–March 2012). Methods: The Clinical Learning Environment, Supervision and Nurse Teacher scale was translated using the modified direct translation method. Statistical analyses were performed using PASW Statistics 18 and AMOS 18.0.0 software. A multivariate analysis was conducted in order to assess construct validity. Cronbach’s alpha coefficient was used to evaluate instrument reliability. Results: An exploratory factorial analysis identified the five dimensions from the original version, and explained 66.4% of the variance. Confirmatory factor analysis supported the factor structure of the Spanish version of the instrument. Cronbach’s alpha coefficient for the scale was .95, ranging from .80 to .97 for the subscales. Conclusion: This version of the Clinical Learning Environment, Supervision and Nurse Teacher scale instrument showed acceptable psychometric properties for use as an assessment scale in Spanish-speaking countries.

Estimation of water retention curve from mercury intrusion porosimetry and van Genuchten model

The water retention curve (WRC) is a hydraulic characteristic of concrete required for advanced modeling of water (and thus solute) transport in variably saturated, heterogeneous concrete. Unfortunately, determination by a direct experimental method (for example, measuring equilibrium moisture levels of large samples stored in constant humidity cells) is a lengthy process, taking over 2 years for large samples. A surrogate approach is presented in which the WRC is conveniently estimated from mercury intrusion porosimetry (MIP) and validated by water sorption isotherms: The well-known Barrett, Joyner and Halenda (BJH) method of estimating the pore size distribution (PSD) from the water sorption isotherm is shown to complement the PSD derived from conventional MIP. This provides a basis for predicting the complete WRC from MIP data alone. The van Genuchten equation is used to model the combined water sorption and MIP results. It is a convenient tool for describing water retention characteristics over the full moisture content range. The van Genuchten parameter estimation based solely on MIP is shown to give a satisfactory approximation to the WRC, with a simple restriction on one. of the parameters.

Multifocal intraocular lens differentiation using defocus curves

Purpose: To determine the most appropriate analysis technique for the differentiation of multifocal intraocular lens (MIOL) designs using defocus curve assessment of visual capability.Methods:Four groups of fifteen subjects were implanted bilaterally with either monofocal intraocular lenses, refractive MIOLs, diffractive MIOLs, or a combination of refractive and diffractive MIOLs. Defocus curves between -5.0D and +1.5D were evaluated using an absolute and relative depth-of-focus method, the direct comparison method and a new 'Area-of-focus' metric. The results were correlated with a subjective perception of near and intermediate vision. Results:Neither depth-of-focus method of analysis were sensitive enough to differentiate between MIOL groups (p>0.05). The direct comparison method indicated that the refractive MIOL group performed better at +1.00, -1.00 and -1.50 D and worse at -3.00, -3.50, -4.00 and -5.00D compared to the diffractive MIOL group (p

Global optimisation of cold-formed steel sections

In this thesis, standard algorithms are used to carry out the optimisation of cold-formed steel purlins such as zed, channel and sigma sections, which are assumed to be simply supported and subjected to a gravity load. For zed, channel and sigma section, the local buckling, distortional buckling and lateral-torsional buckling are considered respectively herein. Currently, the local buckling is based on the BS 5950-5:1998 and EN 1993-1-3:2006. The distortional buckling is calculated by the direct strength method employing the elastic distortional buckling which is calculated by three available approaches such as Hancock (1995), Schafer and Pekoz (1998), Yu (2005). In the optimisation program, the lateral-torsional buckling based on BS 5950-5:1998, AISI and analytical model of Li (2004) are investigated. For the optimisation program, the programming codes are written for optimisation of channel, zed and sigma beam. The full study has been coded into a computer-based analysis program (MATLAB).

Structural effects in photopolymerized sodium AMPS hydrogels crosslinked with poly(ethylene glycol) diacrylate for use as burn dressings

Synthetic hydrogel polymers were prepared by free radical photopolymerization in aqueous solution of the sodium salt of 2-acrylamido-2-methylpropane sulfonic acid (Na-AMPS). Poly(ethylene glycol) diacrylate (PEGDA) and 4,4'-azo-bis(4-cyanopentanoic acid) were used as the crosslinker and UV-photoinitiator, respectively. The effects of varying the Na-AMPS monomer concentration within the range of 30-50% w/v and the crosslinker concentration within the range of 0.1-1.0% mol (relative to monomer) were studied in terms of their influence on water absorption properties. The hydrogel sheets exhibited extremely high swelling capacities in aqueous media which were dependent on monomer concentration, crosslink density, and the ionic strength and composition of the immersion medium. The effects of varying the number-average molecular weight of the PEGDA crosslinker from = 250 to 700 were also investigated. Interestingly, it was found that increasing the molecular weight and therefore the crosslink length at constant crosslink density decreased both the rate of water absorption and the equilibrium water content. Cytotoxicity testing by the direct contact method with mouse fibroblast L929 cells indicated that the synthesized hydrogels were nontoxic. On the basis of these results, it is considered that photopolymerized Na-AMPS hydrogels crosslinked with PEGDA show considerable potential for biomedical use as dressings for partial thickness burns. This paper describes some structural effects which are relevant to their design as biomaterials for this particular application. © 2013 Copyright Taylor and Francis Group, LLC.

Zr-containing hybrid organic-inorganic mesoporous materials:hydrophobic acid catalysts for biodiesel production

Zirconium-containing periodic mesoporous organosilicas (Zr-PMOs) with varying framework organic content have been synthesized through a direct synthesis method. These materials display the excellent textural properties of the analogous inorganic solid acid Zr-SBA-15 material. However, the substitution of silica by organosilicon species provides a strong hydrophobic character. This substitution leads to meaningful differences in the environment surrounding the zirconium metal sites, leading the modification of the catalytic properties of these materials. Although lower metal incorporation is accomplished in the final materials, leading to a lower population of metal sites, hydrophobisation leads to an impressive beneficial effect on the intrinsic catalytic activity of the zirconium sites in biodiesel production by esterification/transesterification of free fatty acid -containing feedstock. Moreover, the catalytic activity of the highly hybridised materials is hardly affected in presence of large amounts of water, confirming their very good water-tolerance. This makes Zr-PMO materials interesting catalysts for biodiesel production from highly acidic water-containing feedstock. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Application of Volterra Equations to Solve Unit Commitment Problem of Optimised Energy Storage and Generation

Development of reliable methods for optimised energy storage and generation is one of the most imminent challenges in modern power systems. In this paper an adaptive approach to load leveling problem using novel dynamic models based on the Volterra integral equations of the first kind with piecewise continuous kernels. These integral equations efficiently solve such inverse problem taking into account both the time dependent efficiencies and the availability of generation/storage of each energy storage technology. In this analysis a direct numerical method is employed to find the least-cost dispatch of available storages. The proposed collocation type numerical method has second order accuracy and enjoys self-regularization properties, which is associated with confidence levels of system demand. This adaptive approach is suitable for energy storage optimisation in real time. The efficiency of the proposed methodology is demonstrated on the Single Electricity Market of Republic of Ireland and Northern Ireland.

Development and optimization of fluoxetine orally disintegrating tablets using Box-Behnken design

Purpose: To develop and optimise some variables that influence fluoxetine orally disintegrating tablets (ODTs) formulation. Methods: Fluoxetine ODTs tablets were prepared using direct compression method. Three-factor, 3- level Box-Behnken design was used to optimize and develop fluoxetine ODT formulation. The design suggested 15 formulations of different lubricant concentration (X1), lubricant mixing time (X2), and compression force (X3) and then their effect was monitored on tablet weight (Y1), thickness (Y2), hardness (Y3), % friability (Y4), and disintegration time (Y5). Results: All powder blends showed acceptable flow properties, ranging from good to excellent. The disintegration time (Y5) was affected directly by lubricant concentration (X1). Lubricant mixing time (X2) had a direct effect on tablet thickness (Y2) and hardness (Y3), while compression force (X3) had a direct impact on tablet hardness (Y3), % friability (Y4) and disintegration time (Y5). Accordingly, Box-Behnken design suggested an optimized formula of 0.86 mg (X1), 15.3 min (X2), and 10.6 KN (X3). Finally, the prediction error percentage responses of Y1, Y2, Y3, Y4, and Y5 were 0.31, 0.52, 2.13, 3.92 and 3.75 %, respectively. Formula 4 and 8 achieved 90 % of drug release within the first 5 min of dissolution test. Conclusion: Fluoxetine ODT formulation has been developed and optimized successfully using Box- Behnken design and has also been manufactured efficiently using direct compression technique.

Efficient representation of texture details in medical images by fusion of Ripplet and DDCT transformed images

Purpose: To evaluate and compare the performance of Ripplet Type-1 transform and directional discrete cosine transform (DDCT) and their combinations for improved representation of MRI images while preserving its fine features such as edges along the smooth curves and textures. Methods: In a novel image representation method based on fusion of Ripplet type-1 and conventional/directional DCT transforms, source images were enhanced in terms of visual quality using Ripplet and DDCT and their various combinations. The enhancement achieved was quantified on the basis of peak signal to noise ratio (PSNR), mean square error (MSE), structural content (SC), average difference (AD), maximum difference (MD), normalized cross correlation (NCC), and normalized absolute error (NAE). To determine the attributes of both transforms, these transforms were combined to represent the entire image as well. All the possible combinations were tested to present a complete study of combinations of the transforms and the contrasts were evaluated amongst all the combinations. Results: While using the direct combining method (DDCT) first and then the Ripplet method, a PSNR value of 32.3512 was obtained which is comparatively higher than the PSNR values of the other combinations. This novel designed technique gives PSNR value approximately equal to the PSNR’s of parent techniques. Along with this, it was able to preserve edge information, texture information and various other directional image features. The fusion of DDCT followed by the Ripplet reproduced the best images. Conclusion: The transformation of images using Ripplet followed by DDCT ensures a more efficient method for the representation of images with preservation of its fine details like edges and textures.

Dynamical modeling of biogas production supported by an experimental anaerobic digestion process

This paper presents the development of a combined experimental and numerical approach to study the anaerobic digestion of both the wastes produced in a biorefinery using yeast for biodiesel production and the wastes generated in the preceding microbial biomass production. The experimental results show that it is possible to valorise through anaerobic digestion all the tested residues. In the implementation of the numerical model for anaerobic digestion, a procedure for the identification of its parameters needs to be developed. A hybrid search Genetic Algorithm was used, followed by a direct search method. In order to test the procedure for estimation of parameters, first noise-free data was considered and a critical analysis of the results obtain so far was undertaken. As a demonstration of its application, the procedure was applied to experimental data.

A Near-Infrared Reflectance Spectroscopy Method for Direct Analysis of Several Chemical Components and Properties of Fruit, for Example, Chinese Hawthorn

Near-infrared spectroscopy (NIRS) calibrations were developed for the discrimination of Chinese hawthorn (Crataegus pinnatifida Bge. var. major) fruit from three geographical regions as well as for the estimation of the total sugar, total acid, total phenolic content, and total antioxidant activity. Principal component analysis (PCA) was used for the discrimination of the fruit on the basis of their geographical origin. Three pattern recognition methods, linear discriminant analysis, partial least-squares-discriminant analysis, and back-propagation artificial neural networks, were applied to classify and compare these samples. Furthermore, three multivariate calibration models based on the first derivative NIR spectroscopy, partial least-squares regression, back-propagation artificial neural networks, and least-squares-support vector machines, were constructed for quantitative analysis of the four analytes, total sugar, total acid, total phenolic content, and total antioxidant activity, and validated by prediction data sets.

Direct precipitation of lead zirconate titanate by the hydrothermal method

Direct precipitation of fine powders of lead zirconate titanate (PZT) in the complete range of solid solution, is investigated under hydrothermal conditions, starting from lead oxide and titania/zirconia mixed gels. The perovskite phase is formed in the temperature range of 165 – 340°C. Sequence of the hydrothermal reactions is studied by identifying the intermediate phases. The initial formation of PbO: TiO2 solid solution is followed by the reaction of the same with the remaining mixed gels giving rise to X-ray amorphous PZT phase. Further, through crystallite growth, the X-ray crystalline PZT is formed. This method can be extended for the preparation of PLZT powder as well. The resulting powders are sinterable to high density ceramics.