Diffusion in a sol-gel-derived medium with a view toward biosensor applications

Time-resolved fluorescence anisotropy and fluorescence recovery after photobleaching were applied to study the diffusion of dyes and a fluorescence-labeled enzyme in a sol-gel-derived medium. This type of medium exhibits attractive properties such as robustness, low processing temperature, high porosity, large internal surface area, and can act as protective immobilization media for biologically active molecules. This makes it a suitable candidate for biosensor applications. The glasslike nature and good optical quality allows for light addressable entities to be incorporated and accessed using spectroscopy. This type of matrix, once formed, can be anything from an ordered gel to a robust glassy block depending on the aging process. In this work we apply confocal microscopy and time-resolved fluorescence techniques to study both rotational and lateral diffusion with aging time within a silica sol-gel derived monolith. An enzyme, horseradish peroxidase, was labeled with Alexa Fluor 488 and rotation related to both the enzyme and the probe monitored during the matrix aging process. Diffusion coefficients of between ca. 0.5 x 10(-7) and 4 x 10(-7) cm2 s(-1) were obtained from preliminary FRAP measurements of fluorescein and correlated to differences in the catalytic activity of HRP incorporated in the monolith.     

Validation of a high-performance chromatographic method for determination of cefotaxime in biological samples

An analytical method for detecting and quantifying cefotaxime in plasma and several tissues is described. The method was developed and validated using plasma and tissues of rats. The samples were analyzed by reversed phase liquid chromatography (HPLC) with UV detection (254 nm). Calibration graphs showed a linear correlation (r > 0.999) over the concentration ranges of 0.5–200 μg/mL and 1.25–25 μg/g for plasma and tissues, respectively. The recovery of cefotaxime from plasma standards prepared at the concentrations of 25 μg/mL and 100 μg/mL was 98.5 ± 3.5% and 101.8 ± 2.2%, respectively. The recovery of cefotaxime from tissue standards of liver, fat and muscle, prepared at the concentration of 10 μg/g was: 89.8 ± 1.2% (liver), 103.9 ± 6.5% (fat) and 97.8 ± 2.1% (muscle). The detection (LOD) and quantitation (LOQ) limits for plasma samples were established at 0.11 μg/mL and 0.49 μg/mL, respectively. The values of these limits for tissues samples were approximately 2.5 times higher: 0.3 μg/g (LOD) and 1.25 μg/g (LOQ). For plasma samples, the deviation of the observed concentration from the nominal concentration was less than 5% and the coefficient of variation for within-day and between-day assays was less than 6% and 12%, respectively. The method was used in a pharmacokinetic study of cefotaxime in the rat and the mean values of the pharmacokinetic parameters are given. Received: 25 May 1998 / Revised: 27 July 1998 / Accepted: 1 August 1998     

Determination of total nitrogen in food by flow injection analysis (FIA) with a potentiometric differential detection system

A flow injection set-up based on potentiometric detection and gas diffusion device for the determination of total nitrogen in food is described. The detection system consisted of two ammonium-sensitive electrodes placed sequentially and each alternately operating as reference electrode. Tubular electrodes without an inner reference solution were prepared with a PVC membrane composed of nonactin in Tris (2-ethylhexyl) phosphate and potassium tetrakis (4-chlorophenyl) borate to reduce the membrane resistance. The food sample digests were inserted into the system, and the ammonium present was converted into ammonia gas. The gas diffused through a gas-permeable membrane to a buffer acceptor stream with a pH that ensured transformation to the ammonium cation, which was potentiometrically detected. Good agreement between FIA results and those provided by the reference procedure was obtained, with relative deviation errors below 5%. Using the proposed system, low reagent consumption is possible, a sampling rate of about 30 samples/h was achieved, as well as a good reproducibility for consecutive injections of the same sample (variation coefficient < 2%). Received: 8 October 1998 / Revised: 7 January 1999 / Accepted: 12 January 1999     

Chemical Analysis and In Vitro Bioactivity of Essential Oil of Laurelia sempervirens and Safrole Derivatives against Oomycete Fish Pathogens

In this study, the essential oil (EO) from Laurelia sempervirens was analyzed by GC/MS and safrole (1) was identified as the major metabolite 1, was subjected to direct reactions on the oxygenated groups in the aromatic ring and in the side chain, and eight compounds (4 to 12) were obtained by the process. EO and compounds 4–12 were subjected to biological assays on 24 strains of the genus Saprolegnia, specifically of the species 12 S. parasitica and 12 S. australis. EO showed a significant effect against Saprolegnia strains. Compound 6 presents the highest activity against two resistant strains, with minimum inhibitory concentration (MIC) and minimum oomyceticidal concentration (MOC) values of 25 to 100 and 75 to 125 µg/mL, respectively. The results show that compound 6 exhibited superior activities compared to the commercial controls bronopol and azoxystrobin used to combat these pathogens.     

Achieving reliability of calculations for flat potential surfaces in density functional theory: The case of Al4 and Al4+1

Harmonic frequencies obtained by finite-differences from nonlocal density functional calculations are presented for the ground states of Al₄ (neutral and cationic). The effect of varying the step size used in the finite-difference evaluation and the influence of the density convergence threshold are discussed. Potential energy curves along the most important normal coordinates are shown. With these results, we found that for Al₄ the square and the rhombus minima are almost degenerate with each other, while for Al₄⁺¹, the rhombus is more stable and the square is a transition state. © 1997 John Wiley & Sons, Inc.     

An approximated principal component prediction model for continuous-time stochastic processes

In this paper, a linear model for forecasting a continuous-time stochastic process in a future interval in terms of its evolution in a past interval is developed. This model is based on linear regression of the principal components in the future against the principal components in the past. In order to approximate the principal factors from discrete observations of a set of regular sample paths, cubic spline interpolation is used. An application for forecasting tourism evolution in Granada is also included. © 1997 by John Wiley & Sons, Ltd.     

In Vitro Antioxidant,Antitumor and Photocatalytic Activities of Silver Nanoparticles Synthesized Using Equisetum Species: A Green Approach

The ethanolic extracts of three Equisetum species (E. pratense Ehrh., E. sylvaticum L. and E. telmateia Ehrh.) were used to reduce silver ions to silver nanoparticles (AgNPs). The synthesized AgNPs were characterized using UV-Vis spectrophotometry, Fourier Transform Infrared Spectroscopy (FTIR), Energy Dispersive X-ray (EDX), Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS) measurements. FTIR data revealed the functional groups of biomolecules involved in AgNPs synthesis, such as O-H, C-H, C=O, C-O, and C-C. EDX spectroscopy was used to highlight the presence of silver, while DLS spectroscopy provided information on the mean diameter of AgNPs, that ranged from 74.4 to 314 nm. The negative Zeta potential values (−23.76 for Ep–AgNPs, −29.54 for Es–AgNPs and −20.72 for Et–AgNPs) indicate the stability of the obtained colloidal solution. The study also focused on establishing the photocatalytic activity of AgNPs, which is an important aspect in terms of removing organic dyes from the environment. The best photocatalytic activity was observed for AgNPs obtained from E. telmateia, which degraded malachite green in a proportion of 97.9%. The antioxidant action of the three AgNPs samples was highlighted comparatively through four tests, with the best overall antioxidant capacity being observed for AgNPs obtained using E. sylvaticum. Moreover, the biosynthesized AgNPs showed promising cytotoxic efficacy against cancerous cell line MG63, the AgNPs obtained from E. sylvaticum L. providing the best result, with a LD₅₀ value around 1.5 mg/mL.