On reducing test length for FSMs with extra states

A long-standing problem when testing from a deterministic finite state machine is to guarantee full fault coverage even if the faults introduce extra states in the implementations. It is well known that such tests should include the sequences in a traversal set which contains all input sequences of length defined by the number of extra states. This paper suggests the SPY method, which helps reduce the length of tests by distributing sequences of the traversal set and reducing test branching. It is also demonstrated that an additional assumption about the implementation under test relaxes the requirement of the complete traversal set. The results of the experimental comparison of the proposed method with an existing method indicate that the resulting reduction can reach 40%. Experimental results suggest that the additional assumption about the implementation can help in further reducing the test suite length. Copyright (C) 2011 John Wiley & Sons, Ltd.

Very Intense Distinct Blue and Red Photoluminescence Emission in MgTiO3 Thin Films Prepared by the Polymeric Precursor Method: An Experimental and Theoretical Approach

MgTiO3 (MTO) thin films were prepared by the polymeric precursor method with posterior spin-coating deposition. The films were deposited on Pt(111)/Ti/SiO2/Si(100) substrates and heat treated at 350 degrees C for 2 h and then heat treated at 400, 450, 500, 550, 600, 650 and 700 C for 2 h. The degree of structural order disorder, optical properties, and morphology of the MTO thin films were investigated by X-ray diffraction (XRD), micro-Raman spectroscopy (MR), ultraviolet-visible (UV-vis) absorption spectroscopy, photoluminescence (PL) measurements, and field-emission gun scanning electron microscopy (FEG-SEM) to investigate the morphology. XRD revealed that an increase in the annealing temperature resulted in a structural organization of MTO thin films. First-principles quantum mechanical calculations based on density functional theory (B3LYP level) were employed to study the electronic structure of ordered and disordered asymmetric models. The electronic properties were analyzed, and the relevance of the present theoretical and experimental results was discussed in the light of PL behavior. The presence of localized electronic levels and a charge gradient in the band gap due to a break in the symmetry are responsible for the PL in disordered MTO lattice.

Selective Permeation and Organic Extraction of Recombinant Green Fluorescent Protein (gfpuv) from Escherichia coli

Background Transformed cells of Escherichia coli DH5-α with pGFPuv, induced by IPTG (isopropyl-β-d-thiogalactopyranoside), express the green fluorescent protein (gfpuv) during growth phases. E. coli subjected to the combination of selective permeation by freezing/thawing/sonication cycles followed by the three-phase partitioning extraction (TPP) method were compared to the direct application of TPP to the same culture of E. coli on releasing gfpuv from the over-expressing cells. Material and Methods Cultures (37°C/100 rpm/ 24 h; μ = 0.99 h-1 - 1.10 h-1) of transformed (pGFP) Escherichia coli DH5-α, expressing the green fluorescent protein (gfpuv, absorbance at 394 nm and emission at 509 nm) were sonicated in successive intervals of sonication (25 vibrations/pulse) to determine the maximum amount of gfpuv released from the cells. For selective permeation, the transformed previously frozen (-75°C) cells were subjected to three freeze/thaw (-20°C/ 0.83°C/min) cycles interlaid by sonication (3 pulses/ 6 seconds/ 25 vibrations). The intracellular permeate with gfpuv in extraction buffer (TE) solution (25 mM Tris-HCl, pH 8.0, 1 mM β-mercaptoethanol β-ME, 0.1 mM PMSF) was subjected to the three-phase partitioning (TPP) method with t-butanol and 1.6 M ammonium sulfate. Sonication efficiency was verified on the application to the cells previously treated by the TPP method. The intra-cell releases were mixed and eluted through methyl HIC column with a buffer solution (10 mM Tris-HCl, 10 mM EDTA, pH 8.0). Results The sonication maximum released amount obtained from the cells was 327.67 μg gfpuv/mL (20.73 μg gfpuv/mg total proteins – BSA), after 9 min of treatment. Through the selective permeation by three repeated freezing/thawing/sonication cycles applied to the cells, a close content of 241.19 μg gfpuv/mL (29.74 μg gfpuv/mg BSA) was obtained. The specific mass range of gfpuv released from the same cultures, by the three-phase partitioning (TPP) method, in relation to total proteins, was higher, between 107.28 μg/mg and 135.10 μg/mg. Conclusions The selective permeation of gfpuv by freezing/thawing/sonication followed by TPP separation method was equivalent to the amount of gfpuv extracted from the cells directly by TPP; although selective permeation extracts showed better elution through the HIC column.

Computer game-based and traditional learning method: a comparison regarding students’ knowledge retention

Abstract Background Educational computer games are examples of computer-assisted learning objects, representing an educational strategy of growing interest. Given the changes in the digital world over the last decades, students of the current generation expect technology to be used in advancing their learning requiring a need to change traditional passive learning methodologies to an active multisensory experimental learning methodology. The objective of this study was to compare a computer game-based learning method with a traditional learning method, regarding learning gains and knowledge retention, as means of teaching head and neck Anatomy and Physiology to Speech-Language and Hearing pathology undergraduate students. Methods Students were randomized to participate to one of the learning methods and the data analyst was blinded to which method of learning the students had received. Students’ prior knowledge (i.e. before undergoing the learning method), short-term knowledge retention and long-term knowledge retention (i.e. six months after undergoing the learning method) were assessed with a multiple choice questionnaire. Students’ performance was compared considering the three moments of assessment for both for the mean total score and for separated mean scores for Anatomy questions and for Physiology questions. Results Students that received the game-based method performed better in the pos-test assessment only when considering the Anatomy questions section. Students that received the traditional lecture performed better in both post-test and long-term post-test when considering the Anatomy and Physiology questions. Conclusions The game-based learning method is comparable to the traditional learning method in general and in short-term gains, while the traditional lecture still seems to be more effective to improve students’ short and long-term knowledge retention.

Equilibrium topology for plasmas with reversed current density.

Actually, transition from positive to negative plasma current and quasi-steady-state alternated current (AC) operation have been achieved experimentally without loss of ionization. The large transition times suggest the use of MHD equilibrium to model the intermediate magnetic field configurations for corresponding current density reversals. In the present work we show, by means of Maxwell equations, that the most robust equilibrium for any axisymmetric configuration with reversed current density requires the existence of several nonested families of magnetic surfaces inside the plasma. We also show that the currents inside the nonested families satisfy additive rules restricting the geometry and sizes of the axisymmetric magnetic islands; this is done without restricting the equilibrium through arbitrary functions. Finally, we introduce a local successive approximations method to describe the equilibrium about an arbitrary reversed current density minimum and, consequently, the transition between different nonested topologies is understood in terms of the eccentricity of the toroidal current density level sets.

Reliable estimation of dense optical flow fields with large displacements

[EN] In this paper we show that a classic optical flow technique by Nagel and Enkelmann can be regarded as an early anisotropic diffusion method with a diffusion tensor. We introduce three improvements into the model formulation that avoid inconsistencies caused by centering the brightness term and the smoothness term in different images use a linear scale-space focusing strategy from coarse to fine scales for avoiding convergence to physically irrelevant local minima, and create an energy functional that is invariant under linear brightness changes.  Applying a gradient descent method to the resulting energy functional leads to a system of diffusion-reaction equations. We prove that this system has a unique solution under realistic assumptions on the initial data, and we present an efficient linear implicit numerical scheme in detail. Our method creates flow fields with 100% density over the entire image domain, it is robust under a large range of parameter variations, and it can recover displacement fields that are far beyond the typical one-pixel limits which are characteristic for many differential methods for determining optical flow. We show that it performs better than the classic optical flow methods with 100%  density that are evaluated by Barron et al. (1994). Our software is available from the Internet.

Development of a fluorescent method for the detection in marine organisms of the respiratory electron transport system

Trabajo realizado por: Maldonado, F.; Packard, T.; Gómez, M.; Santana Rodríguez, J. J

Entwicklung und Evaluierung einer Modellhierarchie zur Simulation der aktinischen Strahlung in aerosolbelasteter und bewölkter Atmosphäre

Das Verstaendnis der in der Atmosphaere stattfindendenOzonproduktions- und Ozonabbaumechanismen ist eines derwichtigsten Ziele der aktuellen Umweltforschung. Da Ozon inder Atmosphaere ausschliesslich durch photochemischeProzesse gebildet wird, ist die Kenntnis der aktinischenStrahlung eine grundlegende Voraussetzung fuer die Prognosevon Ozonkonzentrationen. Die vorliegende Arbeit befasst sichdaher mit der Modellierung der aktinischen Strahlung alsBasis zur Untersuchung photochemischer Prozesse. Dazu wurde eine Modellhierarchie entwickelt, welcheausgehend von der Berechnung der optischen Eigenschaften deratmosphaerischen Extingenten den Strahlungstransport in derAtmosphaere bestimmt. Dazu wurden verschiedeneStrahlungstransportmodelle verwendet, welche sich durch dieLoesungsmethode der Strahlungstransportgleichung und dieAnzahl der raeumlichen Dimensionen unterscheiden. Zur Ueberpruefung der entwickelten Modellhierarchie und zurUntersuchung des Einflusses der verschiedenen Extingentenauf das atmosphaerische Strahlungsfeld wurden zahlreicheSensitivitaetsstudien durchgefuehrt. Dabei zeigte sich, dasssich der Einfluss von Ozon fast ausschliesslich auf denAnteil der Strahlung mit Wellenlaengen kleiner 340 nmauswirkt. Bei der Untersuchung des Einflusses vonAerosolpartikeln und Wolkentropfen auf die aktinischeStrahlung wurde festgestellt, dass die chemischeZusammensetzung, das Vertikalprofil, die Groesse und dieGroessenverteilung der Partikel einen entscheidendenEinfluss nehmen. Eine Evaluierung des 1-dimensionalenStrahlungstransportmodells an Hand von Vergleichen mit denErgebnissen anderen Strahlungstransportmodelle undverschiedenen Messungen ergab, dass die entwickelteModellhierarchie sehr zuverlaessige Resultate liefert. Die Effekte durchbrochener Bewoelkung konnten mit einer2-dimensionalen Simulation untersucht werden. DieUntersuchungen zeigen, dass die haeufig durchgefuehrteapproximative Betrachtung von partieller Bewoelkung mit1-dimensionalen Strahlungstransportmodellen im Vergleich zuden Ergebnissen der 2-dimensionalen Simulation grosse Fehlerverursacht, da die Effekte an den seitlichen Wolkenraendernvon den 1-dimensionalen Strahlungstransportmodellen nichtberuecksichtigt werden koennen.

Study of innovative methods of control in the cereal productive chain for the production of beer and spirits

Researches performed during the PhD course intended to assess innovative applications of near-infrared spectroscopy in reflectance (NIR) in the production chain of beer. The purpose is to measure by NIR the "malting quality" (MQ) parameter of barley, to monitor the malting process and to know if a certain type of barley is suitable for the production of beer and spirits. Moreover, NIR will be applied to monitor the brewing process. First of all, it was possible to check the quality of the raw materials like barley, maize and barley malt using a rapid, non-destructive and reliable method, with a low error of prediction. The more interesting result obtained at this level was that the repeatability of the NIR calibration models developed was comparable with the one of the reference method. Moreover, about malt, new kinds of validation were used in order to estimate the real predictive power of the proposed calibration models and to understand the long-term effects. Furthermore, the precision of all the calibration models developed for malt evaluation was estimated and statistically compared with the reference methods, with good results. Then, new calibration models were developed for monitoring the malting process, measuring the moisture content and other malt quality parameters during germination. Moreover it was possible to obtain by NIR an estimate of the "malting quality" (MQ) of barley and to predict whether if its germination will be rapid and uniform and if a certain type of barley is suitable for the production of beer and spirits. Finally, the NIR technique was applied to monitor the brewing process, using correlations between NIR spectra of beer and analytical parameters, and to assess beer quality. These innovative results are potentially very useful for the actors involved in the beer production chain, especially the calibration models suitable for the control of the malting process and for the assessment of the “malting quality” of barley, which need to be deepened in future studies.

An optimal estimator for the correlation of CMB anisotropies with Large Scale Structures and its application to WMAP-7year and NVSS

In the thesis we present the implementation of the quadratic maximum likelihood (QML) method, ideal to estimate the angular power spectrum of the cross-correlation between cosmic microwave background (CMB) and large scale structure (LSS) maps as well as their individual auto-spectra. Such a tool is an optimal method (unbiased and with minimum variance) in pixel space and goes beyond all the previous harmonic analysis present in the literature. We describe the implementation of the QML method in the {\it BolISW} code and demonstrate its accuracy on simulated maps throughout a Monte Carlo. We apply this optimal estimator to WMAP 7-year and NRAO VLA Sky Survey (NVSS) data and explore the robustness of the angular power spectrum estimates obtained by the QML method. Taking into account the shot noise and one of the systematics (declination correction) in NVSS, we can safely use most of the information contained in this survey. On the contrary we neglect the noise in temperature since WMAP is already cosmic variance dominated on the large scales. Because of a discrepancy in the galaxy auto spectrum between the estimates and the theoretical model, we use two different galaxy distributions: the first one with a constant bias $b$ and the second one with a redshift dependent bias $b(z)$. Finally, we make use of the angular power spectrum estimates obtained by the QML method to derive constraints on the dark energy critical density in a flat $\Lambda$CDM model by different likelihood prescriptions. When using just the cross-correlation between WMAP7 and NVSS maps with 1.8° resolution, we show that $\Omega_\Lambda$ is about the 70\% of the total energy density, disfavouring an Einstein-de Sitter Universe at more than 2 $\sigma$ CL (confidence level).

Validation of a new method to assess the long-term Hypothalamic-Pituitary-Adrenal Activity using hair glucocorticoids as biomarkers. - Studies on pigs and laboratory Sprague-Dawley Rats

The evaluation of chronic activity of the hypothalamic-pituitary-adrenal (HPA) axis is critical for determining the impact of chronic stressful situations. The potential use of hair glucocorticoids as a non-invasive, retrospective, biomarker of long term HPA activity is of great interest, and it is gaining acceptance in humans and animals. However, there are still no studies in literature examining hair cortisol concentration in pigs and corticosterone concentration in laboratory rodents. Therefore, we developed and validated, for the first time, a method for measuring hair glucocorticoids concentration in commercial sows and in Sprague-Dawley rats. Our preliminary data demonstrated: 1) a validated and specific washing protocol and extraction assay method with a good sensitivity in both species; 2) the effect of the reproductive phase, housing conditions and seasonality on hair cortisol concentration in sows; 3) similar hair corticosterone concentration in male and female rats; 4) elevated hair corticosterone concentration in response to chronic stress manipulations and chronic ACTH administration, demonstrating that hair provides a good direct index of HPA activity over long periods than other indirect parameters, such adrenal or thymus weight. From these results we believe that this new non-invasive tool needs to be applied to better characterize the overall impact in livestock animals and in laboratory rodents of chronic stressful situations that negatively affect animals welfare. Nevertheless, further studies are needed to improve this methodology and maybe to develop animal models for chronic stress of high interest and translational value in human medicine.

A mesoscopic simulation method for electrolyte solutions

This thesis deals with the development of a novel simulation technique for macromolecules in electrolyte solutions, with the aim of a performance improvement over current molecular-dynamics based simulation methods. In solutions containing charged macromolecules and salt ions, it is the complex interplay of electrostatic interactions and hydrodynamics that determines the equilibrium and non-equilibrium behavior. However, the treatment of the solvent and dissolved ions makes up the major part of the computational effort. Thus an efficient modeling of both components is essential for the performance of a method. With the novel method we approach the solvent in a coarse-grained fashion and replace the explicit-ion description by a dynamic mean-field treatment. Hence we combine particle- and field-based descriptions in a hybrid method and thereby effectively solve the electrokinetic equations. The developed algorithm is tested extensively in terms of accuracy and performance, and suitable parameter sets are determined. As a first application we study charged polymer solutions (polyelectrolytes) in shear flow with focus on their viscoelastic properties. Here we also include semidilute solutions, which are computationally demanding. Secondly we study the electro-osmotic flow on superhydrophobic surfaces, where we perform a detailed comparison to theoretical predictions.

Development of an LC-MS/MS method for the quantitation of 55 compounds prescribed in combined cardiovascular therapy

This paper reports an LC-MS/MS method with positive electrospray ionization for the screening of commonly prescribed cardiovascular drugs in human plasma, including compounds with antihypertensive (57), antidiabetic (12), hypolipemiant (5), anticoagulant (2) and platelet anti-aggregation (2) effects. Sample treatment consisted of a simple protein precipitation with MeOH/0.1 M ZnSO₄ (4:1, v/v) solution after the addition of internal standard, followed by evaporation and reconstitution. Analytes separation was performed on a Polar-RP column (150 m x 2 mm, 4 μm) using a gradient elution of 15 min. The MS system was operated in MRM mode, monitoring one quantitation and one confirmation transition for each analyte. The recovery of the protein precipitation step ranged from 50 to 70% for most of the compounds, while some were considerably affected by matrix effects. Since several analytes fulfilled the linearity, accuracy and precision values required by the ICH guidelines, the method proved to be suitable for their quantitative analysis. The limits of quantitation varied from 0.38 to 9.1 μg/L and the limits of detection from 0.12 to 5.34 μg/L. The method showed to be suitable for the detection of plasma samples of patients under cardiovascular treatment with the studied drugs, and for 55 compounds reliable quantitative results could be obtained.

LC-MS/MS screening method for designer amphetamines, tryptamines, and piperazines in serum

Since the late 1990s and early 2000s, derivatives of well-known designer drugs as well as new psychoactive compounds have been sold on the illicit drug market and have led to intoxications and fatalities. The LC-MS/MS screening method presented covers 31 new designer drugs as well as cathinone, methcathinone, phencyclidine, and ketamine which were included to complete the screening spectrum. All but the last two are modified molecular structures of amphetamine, tryptamine, or piperazine. Among the amphetamine derivatives are cathinone, methcathinone, 3,4-DMA, 2,5-DMA, DOB, DOET, DOM, ethylamphetamine, MDDMA, 4-MTA, PMA, PMMA, 3,4,5-TMA, TMA-6 and members of the 2C group: 2C-B, 2C-D, 2C-H, 2C-I, 2C-P, 2C-T-2, 2C-T-4, and 2C-T-7. AMT, DPT, DiPT, MiPT, DMT, and 5MeO-DMT are contained in the tryptamine group, BZP, MDBP, TFMPP, mCPP, and MeOPP in the piperazine group. Using an Applied Biosystems LC-MS/MS API 365 TurboIonSpray it is possible to identify all 35 substances. After addition of internal standards and mixed-mode solid-phase extraction the analytes are separated using a Synergi Polar RP column and gradient elution with 1 mM ammonium formate and methanol/0.1% formic acid as mobile phases A and B. Data acquisition is performed in MRM mode with positive electro spray ionization. The assay is selective for all tested substances. Limits of detection were determined by analyzing S/N-ratios and are between 1.0 and 5.0 ng/mL. Matrix effects lie between 65% and 118%, extraction efficiencies range from 72% to 90%.

Cardiogoniometric parameters for detection of coronary artery disease at rest as a function of stenosis localization and distribution

Cardiogoniometry (CGM), a spatiotemporal electrocardiologic 5-lead method with automated analysis, may be useful in primary healthcare for detecting coronary artery disease (CAD) at rest. Our aim was to systematically develop a stenosis-specific parameter set for global CAD detection. In 793 consecutively admitted patients with presumed non-acute CAD, CGM data were collected prior to elective coronary angiography and analyzed retrospectively. 658 patients fulfilled the inclusion criteria, 405 had CAD verified by coronary angiography; the 253 patients with normal coronary angiograms served as the non-CAD controls. Study patients--matched for age, BMI, and gender--were angiographically assigned to 8 stenosis-specific CAD categories or to the controls. One CGM parameter possessing significance (P < .05) and the best diagnostic accuracy was matched to one CAD category. The area under the ROC curve was .80 (global CAD versus controls). A set containing 8 stenosis-specific CGM parameters described variability of R vectors and R-T angles, spatial position and potential distribution of R/T vectors, and ST/T segment alterations. Our parameter set systematically combines CAD categories into an algorithm that detects CAD globally. Prospective validation in clinical studies is ongoing.