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.     

Functional analysis of the cysteine residues and the repetitive sequence of Saccharomyces cerevisiae Pir4/Cis3: the repetitive sequence is needed for binding to the cell wall beta-1,3-glucan

Identification of PIR/CIS3 gene was carried out by amino-terminal sequencing of a protein band released by beta-mercaptoethanol (beta-ME) from S. cerevisiae mnn9 cell walls. The protein was released also by digestion with beta-1,3-glucanases (laminarinase or zymolyase) or by mild alkaline solutions. Deletion of the two carboxyterminal Cys residues (Cys(214)-12aa-Cys(227)-COOH), reduced but did not eliminate incorporation of Pir4 (protein with internal repeats) by disulphide bridges. Similarly, site-directed mutation of two other cysteine amino acids (Cys(130)Ser or Cys(197)Ser) failed to block incorporation of Pir4; the second mutation produced the appearance of Kex2-unprocessed Pir4. Therefore, it seems that deletion or mutation of individual cysteine molecules does not seem enough to inhibit incorporation of Pir4 by disulphide bridges. In fks1Delta and gsc2/fks2Delta cells, defective in beta-1,3-glucan synthesis, modification of the protein pattern found in the supernatant of the growth medium, as well as the material released by beta-ME or laminarinase, was evident. However, incorporation of Pir4 by both disulphide bridges and to the beta-1,3-glucan of the cell wall continued. Deletion of the repetitive sequence (QIGDGQVQA) resulted in the secretion and incorporation by disulphide bridges of Pir4 in reduced amounts together with substantial quantities of the Kex2-unprocessed Pir4 form. Pir4 failed to be incorporated in alkali-sensitive linkages involving beta-1,3-glucan when the first repetitive sequence was deleted. Therefore, this suggests that this sequence is needed in binding Pir4 to the beta-1,3-glucan.     

Separation of recombinant human erythropoietin glycoforms by capillary electrophoresis using volatile electrolytes. Assessment of mass spectrometry for the characterization of erythropoietin glycoforms

The separation of the glycoforms of erythropoietin (EPO) by capillary electrophoresis (CE) was recently published as a monograph by the European Pharmacopoeia (European Pharmacopoeia 4 2002, 1316, 1123-1128). Although the experimental CE conditions employed a background electrolyte containing additives suitable for on-line UV-absorption detection, they were not appropriate for on-line mass spectrometry (MS) detection. In this work, an attempt was made to investigate experimental conditions employing volatile electrolyte systems to achieve the separation and characterization of EPO glycoforms using CE and ESI-MS methodologies. The influence of several operating conditions, such as the coating of the internal walls of the capillary as well as the composition, concentration, and the pH of the separation buffer were investigated. The results demonstrated that when the internal walls of the capillaries were permanently coated with Polybrene and a buffer electrolyte containing 400 mM of HAc-NH4Ac (acetic acid-ammonium acetate), pH 4.75, was used, a significantly reproducible separation was achieved for EPO glycoforms. Intact EPO was characterized by two mass spectrometry techniques: electrospray ionization (ESI-MS) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-MS). The data demonstrated that MALDI-TOF-MS provided a good approximation to an average molecular mass of the EPO molecule. However, it was still necessary to carry out further separation of the intact EPO glycoforms in order to obtain molecular mass information when ESI-MS was used.     

Adaptive value function approximation for continuous-state stochastic dynamic programming

Approximate dynamic programming (ADP) commonly employs value function approximation to numerically solve complex dynamic programming problems. A statistical perspective of value function approximation employs a design and analysis of computer experiments (DACE) approach, where the “computer experiment” yields points on the value function curve. The DACE approach has been used to numerically solve high-dimensional, continuous-state stochastic dynamic programming, and performs two tasks primarily: (1) design of experiments and (2) statistical modeling. The use of design of experiments enables more efficient discretization. However, identifying the appropriate sample size is not straightforward. Furthermore, identifying the appropriate model structure is a well-known problem in the field of statistics. In this paper, we present a sequential method that can adaptively determine both sample size and model structure. Number-theoretic methods (NTM) are used to sequentially grow the experimental design because of their ability to fill the design space. Feed-forward neural networks (NNs) are used for statistical modeling because of their adjustability in structure-complexity . This adaptive value function approximation (AVFA) method must be automated to enable efficient implementation within ADP. An AVFA algorithm is introduced, that increments the size of the state space training data in each sequential step, and for each sample size a successive model search process is performed to find an optimal NN model. The new algorithm is tested on a nine-dimensional inventory forecasting problem.