HPLC confirmatory method development for the determination of seven quinolones in salmon tissue (Salmo salar L.) validated according to the European Union Decision 2002/657/EC

A confirmatory high pressure liquid chromatographic method for the determination of seven quinolone antibiotics in tissue of Atlantic salmon (Salmo salar L.) was developed. Ciprofloxacin (CIP), danofloxacin (DAN), enrofloxacin (ENR), sarafloxacin (SAR), oxolinic acid (OXO), nalidixic acid (NAL) and flumequine (FLU) were separated on a Perfectsil ODS-2 120 (250 mm × 4 mm, 5 μm) column by gradient elution with a mobile phase consisting of 0.1% trifluoroacetic acid (pH = 1), acetonitrile and methanol at 25 °C within 22 min. Analytes were monitored at 255 nm (for the determination of OXO, NAL and FLU) and 275 nm (for CIP, DAN, ENR and SAR) by means of photodiode array detector. Examined quinolones were isolated from salmon tissue by extraction with citrate buffer solution (pH = 4.7) and purified by solid phase extraction using Oasis HLB (200 mg/6 mL) cartridges. The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability and sensitivity according to the European Union Decision 2002/657/EC. The accuracy of the method was additionally proved by its application to certified reference material of salmon tissue (BCR® 725).     

Natural and human forcing in recent geomorphic change; case studies in the Rio de la Plata basin

An analysis of geomorphic system's response to change in human and natural drivers in some areas within the Río de la Plata basin is presented. The aim is to determine whether an acceleration of geomorphic processes has taken place in recent years and, if so, to what extent it is due to natural (climate) or human (land-use) drivers. Study areas of different size, socio-economic and geomorphic conditions have been selected: the Río de la Plata estuary and three sub-basins within its watershed. Sediment cores were extracted and dated (²¹⁰Pb) to determine sedimentation rates since the end of the 19th century. Rates were compared with time series on rainfall as well as human drivers such as population, GDP, livestock load, crop area, energy consumption or cement consumption, all of them related to human capacity to disturb land surface. Data on river discharge were also gathered. Results obtained indicate that sedimentation rates during the last century have remained essentially constant in a remote Andean basin, whereas they show important increases in the other two, particularly one located by the São Paulo metropolitan area. Rates in the estuary are somewhere in between. It appears that there is an intensification of denudation/sedimentation processes within the basin.Rainfall remained stable or varied very slightly during the period analysed and does not seem to explain increases of sedimentation rates observed. Human drivers, particularly those more directly related to capacity to disturb land surface (GDP, energy or cement consumption) show variations that suggest human forcing is a more likely explanation for the observed change in geomorphic processes. It appears that a marked increase in denudation, of a “technological” nature, is taking place in this basin and leading to an acceleration of sediment supply. This is coherent with similar increases observed in other regions.     

Computational evolutionary inverse lagrangian puff model

This article presents a model based on the principles of evolutionary computation and on the principles of dispersion of an Inverse lagrangian puff model of the type Backward gaussian puff tracking in order to determine the spatial behavior over time of the concentration for PM_2.5 and PM₁₀ in a study area.     

Developing BP-driven web applications through the use of MDE techniques

Model driven engineering (MDE) is a suitable approach for performing the construction of software systems (in particular in the Web application domain). There are different types of Web applications depending on their purpose (i.e., document-centric, interactive, transactional, workflow/business process-based, collaborative, etc). This work focusses on business process-based Web applications in order to be able to understand business processes in a broad sense, from the lightweight business processes already addressed by existing proposals to long-running asynchronous processes. This work presents a MDE method for the construction of systems of this type. The method has been designed in two steps following the MDE principles. In the first step, the system is represented by means of models in a technology-independent manner. These models capture the different aspects of Web-based systems (these aspects refer to behaviour, structure, navigation, and presentation issues). In the second step, the model transformations (both model- to-model and model-to-text) are applied in order to obtain the final system in terms of a specific technology. In addition, a set of Eclipse-based tools has been developed to provide automation in the application of the proposed method in order to validate the proposal.     

Capillary electrophoresis of cytochrome P-450 epoxygenase metabolites of arachidonic acid. 1. Resolution of regioisomers

The essential fatty acid arachidonate is oxidized by cytochrome P-450 epoxygenases to four epoxyeicosatrienoic acids (EETs): 14,15-, 11,12-, 8,9-, and 5,6-EETs. Each of the four EET regioisomers and their hydrolysis products (DHETs) has multiple paracrine and autocrine functions and may also potently dilate blood vessels and activate potassium channels. The present work describes a method to resolve EETs and DHETs by capillary electrophoresis (CE) using trimethyl-beta-cyclodextrin and CH3CN as buffer additives. While stored at 25 degrees C, most of the EET and DHET regioisomers remained intact when suspended in alkaline vehicle. However, under these same conditions, 5,6-EET rapidly broke down to a lactone and was slowly converted to 5,6-DHET. When subjected to CE, the EET and DHET regioisomers were baseline resolved (R > or = 1.3); 10 pg of an EET or a DHET regioisomer was readily detectable at 194 nm. In addition, the UV spectra were regiospecific and identical to those obtained during HPLC except that an additional, weak absorption occurred at 235 nm. Together, the high-sensitivity, high-resolution, and differential UV spectra permitted the identification and quantification of EETs in phospholipids isolated from murine liver. Thus, CE was successfully used for the trace analysis of eicosanoids.     

Development of a system for categorizing client reactions to therapist interventions.

We developed a measure of client reactions to therapist interventions. The 21 categories of the measure were divided into 14 positive and 7 negative reactions, which differed significantly from each other on client helpfulness ratings. Preliminary validity data indicated that therapist intentions were related to client reactions more for successful cases than unsuccessful cases, pretreatment symptomatology was highly predictive of which reactions the clients reported, there were some predictable changes in reactions across time in treatment, and within-case correlations of reactions with client-rated session depth and smoothness indicated some similarities across cases. We discuss the case-specific nature of client reactions, methodological issues, and the need for greater therapist awareness of client reactions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Occurrence of copper resistant mutants in the toxic cyanobacteria Microcystis aeruginosa: characterisation and future implications in the use of copper sulphate as algaecide

Freshwater toxic cyanobacteria are an increasing problem to human and animal health. Control of cyanobacteria in water supply reservoirs involves the use of algaecides, such as copper sulphate, usually in a repetitive way. Repercussions of recurrent algaecide treatments on cyanobacteria population dynamics remain still unknown. We studied the adaptation of cyanobacteria to lethal doses of copper sulphate by using Microcystis aeruginosa as an experimental model. A fluctuation analysis demonstrated that copper-resistant cells arise by spontaneous mutations that occur randomly prior to exposition to copper sulphate. The rate of spontaneous mutation from copper sensitivity to resistance was 1.76×10⁻⁶ mutants per cell division. Resistant mutants exhibited a diminished fitness in the absence of copper sulphate, but only these variants were able to grow at Cu²⁺ concentrations higher than 5.8 μM. In addition, copper-resistant cells were significantly smaller than wild-type ones. Warnings on the long-term consequences of repetitive algaecide treatments in water supplies are suggested.     

Direct electron transfer of cytochrome P450 2B4 at electrodes modified with nonionic detergent and colloidal clay nanoparticles

A method for construction of biosensors with membranous cytochrome P450 isoenzymes was developed based on clay/detergent/protein mixed films. Thin films of sodium montmorillonite colloid with incorporated cytochrome P450 2B4 (CYP2B4) with nonionic detergent were prepared on glassy carbon electrodes. The modified electrodes were electrochemically characterized, and bioelectrocatalytic reactions were followed. CYP2B4 can be reduced fast on clay-modified glassy carbon electrodes in the presence of the nonionic detergent Tween 80. In anaerobic solutions, reversible oxidation and reduction is obtained with a formal potential between -0.292 and -0.305 V vs Ag/AgCl 1 M KCl depending on the preparation of the biosensor. In air-saturated solution, bioelectrocatalytic reduction currents can be obtained with the CYP2B4-modified electrode on addition of typical substrates such as aminopyrine and benzphetamine. This reaction was suppressed when methyrapone, an inhibitor of P450 reactions, was present. Measurement of product formation also indicates the bioelectrocatalysis by CYP2B4.