Double-Tagging Polymerase Chain Reaction with a Thiolated Primer and Electrochemical Genosensing based on Gold Nanocomposite Sensor for Food Safety

A novel material for electrochemical biosensing based on rigid conducting gold nanocomposite (nano-AuGEC) is presented. Islands of chemisorbing material (gold nanoparticles) surrounded by nonreactive, rigid, and conducting graphite epoxy composite are thus achieved to avoid the stringent control of surface coverage parameters required during immobilization of thiolated oligos in continuous gold surfaces. The spatial resolution of the immobilized thiolated DNA was easily controlled by merely varying the percentage of gold nanoparticles in the composition of the composite. As low as 9 fmol (60 pM) of synthetic DNA were detected in hybridization experiments when using a thiolated probe. Moreover, for the first time a double tagging PCR strategy was performed with a thiolated primer for the detection of Salmonella sp., one of the most important foodborne pathogens affecting food safety. Ibis assay was performed by double-labeling the amplicon during the PCR with a -DIG and -SH set of labeled primers. The thiolated end allows the immobilization of the amplicon on the nano-AuGEC electrode, while digoxigenin allows the electrochemical detection with the antiDIG-HRP reporter in the femtomole range. Rigid conducting gold nanocomposite represents a good material for the improved and oriented immobilization of biomolecules with excellent transducing properties for the construction of a wide range of electrochemical biosensors such as immunosensors, genosensors, and enzymosensors.

Visual timing and adaptive behavior in a road-crossing simulation study

In this road-crossing simulation study, we assessed both participant's ability to visually judge whether or not they could cross a road, and their adaptive walking behavior. To this end, participants were presented with a road inside the laboratory on which a bike approached with different velocities from different distances. Eight children aged 5-7, ten children aged 10-12, and ten adults were asked both to verbally judge whether they could cross the road, and to actually walk across the road if possible. The results indicated that the verbal judgments were not similar to judgments to actually cross the road. With respect to safety and accuracy of judgments, groups did not differ from each other, although the youngest group tended to be more cautious. All groups appeared to use a strategy to cross the road based both on the distance and the velocity of the approaching bike. Young children waited longer on the curb before crossing the road than older children and adults. All groups adjusted their crossing time to the time-to-arrival of the bike. These findings are discussed in relation to the ecological psychological approach and the putative dissociation between vision for perception (i.e. verbal judgment) and vision for action (i.e. actual crossing). (c) 2004 Elsevier Ltd. All rights reserved.

Magnetic resonance study of the crystallization behavior of InF3-based glasses doped with Cu2+, Mn2+ and Gd3+

The crystallization of fluoroindate glasses doped with Gd3+, Mn2+ and Cu2+ heat treated at different temperatures, ranging from the glass transition temperature (Tg) to the crystallization temperature (Tc), are investigated by electron paramagnetic resonance (EPR) and 19F nuclear magnetic resonance (NMR). The EPR spectra indicate that the Cu2+ ions in the glass are located in axially distorted octahedral sites. In the crystallized glass, the g-values agreed with those reported for Ba2ZnF6, which correspond to Cu2+ in a tetragonal compressed F- octahedron and to Cu2+ on interstitial sites with a square-planar F- co-ordination. The EPR spectra of the Mn2+ doped glasses exhibit a sextet structure due to the Mn2+ hyperfine interaction. These spectra suggest a highly ordered environment for the Mn2+ ions (close to octahedral symmetry) in the glass. The EPR spectra of the recrystallized sample exhibit resonances at the same position, suggesting that the Mn2+ ions are located in sites of highly symmetric crystalline field. The increase of the line intensity of the sextet and the decrease of the background line in the thermal treated samples suggest that the Mn2+ ions move to the highly ordered sites which contribute to the sextet structure. The EPR spectra of the Gd3+ doped glasses exhibit the typical U-spectrum of a s-state ion in a low symmetry site in disordered systems. The EPR of the crystallized glasses, in contrast, have shown a strong resonance in g ≈ 2.0, suggesting Gd3+ ions in environment close to cubic symmetry. The 19F NMR spin-lattice relaxation rates were also strongly influenced by the crystallization process that takes over in samples annealed above Tc. For the glass samples (doped or undoped) the 19F magnetization recoveries were found to be adjusted by an exponential function and the spin-lattice relaxation was characterized by a single relaxation time. In contrast, for the samples treated above Tc, the 19F magnetization-recovery becomes non-exponential. A remarkable feature of our results is that the changes in the Cu2+, Mn2+, Gd3+ EPR spectra and NMR relaxation, are always observed for the samples annealed above Tc. © 2006 Elsevier B.V. All rights reserved.

New voltammetry-based technique for the determination of tebuthiuron in crystal and brown sugar samples

Sugar is widely consumed worldwide and Brazil is the largest producer, consumer, and exporter of this product. To guarantee proper development and productivity of sugar cane crops, it is necessary to apply large quantities of agrochemicals, especially herbicides and pesticides. The herbicide tebuthiuron (TBH) prevents pre- and post-emergence of infesting weed in sugarcane cultures. Considering that it is important to ensure food safety for the population, this paper proposes a reliable method to analyse TBH in sugar matrixes (brown and crystal) using square wave voltammetry (SWV) and differential pulse voltammetry (DPV) at bare glassy carbon electrode and investigate the electrochemical behavior of this herbicide by cyclic voltammetry (CV). Our results suggest that TBH or the product of its reaction with a supporting electrolyte is oxidized through irreversible transfer of one electron between the analyte and the working electrode, at a potential close to +1.16 V vs. Ag |AgClsat in 0.10 mol L-1 KOH as supporting electrolyte solution. Both DPV and SWV are satisfactory for the quantitative analysis of the analyte. DPV is more sensitive and selective, with detection limits of 0.902, 0.815 and 0.578 mg kg-1, and quantification limits of 0.009, 0.010 and 0.008 mg kg-1 in the absence of the matrix and in the presence of crystal and brown sugar matrix, respectively. Repeatability lay between 0.53 and 13.8%, precision ranged between 4.14 and 15.0%, and recovery remained between 84.2 and 113% in the case of DPV conducted in the absence of matrix and in the presence of the crystal sugar matrix, respectively.

Topical Application of Retinyl Palmitate-Loaded Nanotechnology-Based Drug Delivery Systems for the Treatment of Skin Aging

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fatigue Behavior of Weld Repaired AISI 4130 Aeronautic Steel Used in Critical Flight Safety Structures

Since the 1950s, fatigue is the most important project and operational consideration for both civil and military aircrafts. For some aircraft models the most loaded component is one that supports the motor: the Motor Cradle. Because they are considered critical to the flight safety the aeronautic standards are extremely rigorous in manufacturing them by imposing a zero index of defects on the final weld quality (Safe Life), which is 100% inspected by Non-Destructive Testing/NDT. This study has as objective to evaluate the effects of up to four successive TIG welding repairs on the axial fatigue strength of an AISI 4130 steel. Tests were conducted on hot-rolled steel plate specimens, 0.89 mm thick, with load ratio R = 0.1, constant amplitude, at 20 Hz frequency and in room temperature, in accordance with ASTM E466 Standard. The results were related to microhardness and microstructural and geometric changes resulting from welding cycles.

Metabolic and Behavior Changes in Surubim Acutely Exposed to a Glyphosate-Based Herbicide

This study examined the effect of glyphosate-based herbicide (Roundup Original), the major herbicide used in soybean crops in Mato Grosso state, at concentrations of 0, 2.25, 4.5, 7.5, and 15 mg L-1 on metabolic and behavior parameters of the hybrid fish surubim in an acute exposure lasting 96 h. Glycogen content, glucose, lactate, and protein levels were measured in different tissues. Plasma levels of cholesterol, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were also determined. Ventilatory frequency (VF) and swimming activity (SA) were considered behavior parameters. Results showed that herbicide exposure decreased plasma glucose levels and increased it in surubim liver. Lactate increased in both plasma and liver but decreased in muscle. Protein levels decreased in plasma and muscle but increased in liver. After herbicide exposure, liver and muscle glycogen was decreased. Cholesterol levels decreased in plasma at all concentrations tested. Plasma ALT increased, and no alterations were recorded for AST levels. VF increased after glyphosate exposure (5 min) and decreased after 96 h. SA showed differences among all groups (5 min). At the end of 96 h, SA was altered by the 7.5 mg L-1 concentration. Fish used anaerobic glycolysis as indicated by generally decreased glycogen levels and decreased lactate levels in muscle but increased ones in plasma and liver. We suggest that the studied parameters could be used as indicators of herbicide toxicity in surubim and may provide extremely important information for understanding the biology of the animal and its responsiveness to external stimuli (stressors).

Comparative electrical behavior at low and high current of SnO(2)- and ZnO-based varistors

The complete I-V characteristics of SnO(2)-based varistors, particularly of the Pianaro system SCNCr consisting in 98.9%SnO(2)+1%CoO+0.05%Nb(2)O(5)+0.05%Cr(2)O(3), all in mol%, have been seldom reported in the literature. A comparative study at low and high currents of the nonohmic behavior of SCNCr- and ZnO-based varistors (modified Matsuoka system) is proposed in this work. The SCNCr system showed higher nonlinearity coefficients in the whole range of measured current. The electrical breakdown field (E(b)) was twice as high for the SCNCr system (5400 V/cm) than for the ZnO varistor (2600 V/cm) due to a smaller average grain size of the former (4.5 mu m) with respect to the latter (8.5 mu m). Nevertheless, we consider that another important factor responsible for the high E(b) in the SCNCr system is the great number of electrically active interfaces (85%) as determined with electrostatic force microscopy (EFM). It was also established that the SCNCr system might be produced in disks of smaller dimensions than that of commercial ZnO-based product, with a 5.0 cm(-1) minimal area-volume (A/V) ratio. The SCNCr reached the saturation current in a short time because of the high resistivity of the grains, which is five times higher than that of the grains in ZnO-based varistors.

Vitreous Pharmacokinetics and Retinal Safety of Intravitreal Preserved Versus Non-preserved Triamcinolone Acetonide in Rabbit Eyes

Purpose: To compare the intravitreal pharmacokinetic profile of a triamcinolone acetonide formulation containing the preservative benzyl alcohol (TA-BA) versus a preservative-free triamcinolone acetonide formulation (TA-PF), and evaluate potential signs of toxicity to the retina. Methods: A total of 60 New Zealand male white rabbits, divided into two groups, were studied. In the TA-BA group, 30 rabbits received an intravitreal injection of TA-BA (4 mg/0.1ml) into the right eye. In the TA-PF group, 30 rabbits received an intravitreal injection of TA-PF (4 mg/0.1ml) into the right eye. The intravitreal drug levels were determined in 25 animals from each group by high-performance liquid chromatography (HPLC). The potential for toxicity associated with the intravitreal triamcinolone injections was evaluated in five randomly selected animals from each group by electroretinography (ERG) and by light microscopy. Results: Median intravitreal concentrations of TA-BA (mu g/ml) were 1903.1, 1213.0, 857.8, 442.0, 248.6 at 3, 7, 14, 21 and 28 days after injection. Intravitreal concentrations of TA-PF (mu g/ml) were 1032.9, 570.1, 516.6, 347.9, 102.8 at 3, 7, 14, 21 and 28 days after injection. The median intravitreal triamcinolone concentration was significantly higher in the TA-BA compared to the TA-PF group at 7 days post-injection (p < 0.05). There was no significant difference between the two groups in median triamcinolone concentration at the other time points evaluated. There was no evidence of toxic effects on the retina in either group based on ERG or histological analyses. Conclusions: Following a single intravitreal injection, the median concentration of triamcinolone acetonide is significantly higher in the TA-BA compared to the TA-PF group at 7 days post-injection. No toxic reactions in the retina were observed in either group.

Intervention Based Exclusively on Stage- Matched Printed EducationalMaterials Regarding Healthy Eating Does Not Result in Changes to Adolescents' Dietary Behavior

Purpose. To assess the impact of a six-month stage-based intervention on fruit and vegetable intake, regarding perceived benefits and barriers, and self-efficacy among adolescents. Design. Randomized treatment-control, pre-post design. Subjects/ Setting. Schools were randomized between control and experimental groups. 860 adolescents from ten public schools in Bras ' ilia, Federal District, Brazil were evaluated at baseline; 771 (81%) completed the study. Intervention. Experimental group received monthly magazines and newsletters aimed at promotion of healthy eating. Measures. Self-reported fruit and vegetable intake, stages of change, self-efficacy and decisional balance scores were evaluated at baseline and post-intervention in both groups. Analysis. The effectiveness of the intervention was evaluated using the analysis of covariance model (ANCOVA) and repeated measurement analysis by means of weighted least squares. Comparison between the proportions of adolescents who advanced through the stages during the intervention was performed using the Mantel-Haenszel chi-square test. Results. After adjusting for sex and age, study variables showed no modifications through the proposed intervention. There was no statistical difference in participant mobility in the intervention and control groups between the stages of change, throughout the study. Conclusion. A nutritional intervention based exclusively on distribution of stage-matched printed educational materials was insufficient to change adolescents' dietary behavior.

Shear behavior of reinforced concrete slabs under concentrated load: an investigation through Sequentially Linear Analysis

ABSTRACT (italiano) Con crescente attenzione riguardo al problema della sicurezza di ponti e viadotti esistenti nei Paesi Bassi, lo scopo della presente tesi è quello di studiare, mediante la modellazione con Elementi Finiti ed il continuo confronto con risultati sperimentali, la risposta in esercizio di elementi che compongono infrastrutture del genere, ovvero lastre in calcestruzzo armato sollecitate da carichi concentrati. Tali elementi sono caratterizzati da un comportamento ed una crisi per taglio, la cui modellazione è, da un punto di vista computazionale, una sfida piuttosto ardua, a causa del loro comportamento fragile combinato a vari effetti tridimensionali. La tesi è incentrata sull'utilizzo della Sequentially Linear Analysis (SLA), un metodo di soluzione agli Elementi Finiti alternativo rispetto ai classici approcci incrementali e iterativi. Il vantaggio della SLA è quello di evitare i ben noti problemi di convergenza tipici delle analisi non lineari, specificando direttamente l'incremento di danno sull'elemento finito, attraverso la riduzione di rigidezze e resistenze nel particolare elemento finito, invece dell'incremento di carico o di spostamento. Il confronto tra i risultati di due prove di laboratorio su lastre in calcestruzzo armato e quelli della SLA ha dimostrato in entrambi i casi la robustezza del metodo, in termini di accuratezza dei diagrammi carico-spostamento, di distribuzione di tensioni e deformazioni e di rappresentazione del quadro fessurativo e dei meccanismi di crisi per taglio. Diverse variazioni dei più importanti parametri del modello sono state eseguite, evidenziando la forte incidenza sulle soluzioni dell'energia di frattura e del modello scelto per la riduzione del modulo elastico trasversale. Infine è stato effettuato un paragone tra la SLA ed il metodo non lineare di Newton-Raphson, il quale mostra la maggiore affidabilità della SLA nella valutazione di carichi e spostamenti ultimi insieme ad una significativa riduzione dei tempi computazionali. ABSTRACT (english) With increasing attention to the assessment of safety in existing dutch bridges and viaducts, the aim of the present thesis is to study, through the Finite Element modeling method and the continuous comparison with experimental results, the real response of elements that compose these infrastructures, i.e. reinforced concrete slabs subjected to concentrated loads. These elements are characterized by shear behavior and crisis, whose modeling is, from a computational point of view, a hard challenge, due to their brittle behavior combined with various 3D effects. The thesis is focused on the use of Sequentially Linear Analysis (SLA), an alternative solution technique to classical non linear Finite Element analyses that are based on incremental and iterative approaches. The advantage of SLA is to avoid the well-known convergence problems of non linear analyses by directly specifying a damage increment, in terms of a reduction of stiffness and strength in the particular finite element, instead of a load or displacement increment. The comparison between the results of two laboratory tests on reinforced concrete slabs and those obtained by SLA has shown in both the cases the robustness of the method, in terms of accuracy of load-displacements diagrams, of the distribution of stress and strain and of the representation of the cracking pattern and of the shear failure mechanisms. Different variations of the most important parameters have been performed, pointing out the strong incidence on the solutions of the fracture energy and of the chosen shear retention model. At last a confrontation between SLA and the non linear Newton-Raphson method has been executed, showing the better reliability of the SLA in the evaluation of the ultimate loads and displacements, together with a significant reduction of computational times.

Physical model of PD behavior and relevant damage growth from micro-cavities in polyethylene-based material under AC voltage

In the framework of developing defect-based life models, in which breakdown is explicitly associated with partial discharge (PD)-induced damage growth from a defect, ageing tests and PD measurements were carried out in the lab on polyethylene (PE) layered specimens containing artificial cavities. PD activity was monitored continuously during aging. A quasi-deterministic series of stages can be observed in the behavior of the main PD parameters (i.e. discharge repetition rate and amplitude). Phase-resolved PD patterns at various ageing stages were reproduced by numerical simulation which is based on a physical discharge model devoid of adaptive parameters. The evolution of the simulation parameters provides insight into the physical-chemical changes taking place at the dielectric/cavity interface during the aging process. PD activity shows similar time behavior under constant cavity gas volume and constant cavity gas pressure conditions, suggesting that the variation of PD parameters may not be attributed to the variation of the gas pressure. Brownish PD byproducts, consisting of oxygen containing moieties, and degradation pits were found at the dielectric/cavity interface. It is speculated that the change of PD activity is related to the composition of the cavity gas, as well as to the properties of dielectric/cavity interface.

Investigations on asymptotic safety of metric, tetrad and Einstein-Cartan gravity

Among the different approaches for a construction of a fundamental quantum theory of gravity the Asymptotic Safety scenario conjectures that quantum gravity can be defined within the framework of conventional quantum field theory, but only non-perturbatively. In this case its high energy behavior is controlled by a non-Gaussian fixed point of the renormalization group flow, such that its infinite cutoff limit can be taken in a well defined way. A theory of this kind is referred to as non-perturbatively renormalizable. In the last decade a considerable amount of evidence has been collected that in four dimensional metric gravity such a fixed point, suitable for the Asymptotic Safety construction, indeed exists. This thesis extends the Asymptotic Safety program of quantum gravity by three independent studies that differ in the fundamental field variables the investigated quantum theory is based on, but all exhibit a gauge group of equivalent semi-direct product structure. It allows for the first time for a direct comparison of three asymptotically safe theories of gravity constructed from different field variables. The first study investigates metric gravity coupled to SU(N) Yang-Mills theory. In particular the gravitational effects to the running of the gauge coupling are analyzed and its implications for QED and the Standard Model are discussed. The second analysis amounts to the first investigation on an asymptotically safe theory of gravity in a pure tetrad formulation. Its renormalization group flow is compared to the corresponding approximation of the metric theory and the influence of its enlarged gauge group on the UV behavior of the theory is analyzed. The third study explores Asymptotic Safety of gravity in the Einstein-Cartan setting. Here, besides the tetrad, the spin connection is considered a second fundamental field. The larger number of independent field components and the enlarged gauge group render any RG analysis of this system much more difficult than the analog metric analysis. In order to reduce the complexity of this task a novel functional renormalization group equation is proposed, that allows for an evaluation of the flow in a purely algebraic manner. As a first example of its suitability it is applied to a three dimensional truncation of the form of the Holst action, with the Newton constant, the cosmological constant and the Immirzi parameter as its running couplings. A detailed comparison of the resulting renormalization group flow to a previous study of the same system demonstrates the reliability of the new equation and suggests its use for future studies of extended truncations in this framework.

Sensor-based evaluation of Circadian motor behavior in people with Dementia. Development and validation of analysis strategies

[ITA]La demenza consiste nel deterioramento, spesso progressivo, dello stato cognitivo di un individuo. Chi è affetto da demenza, presenta alterazioni a livello cognitivo, comportamentale e motorio, ad esempio compiendo gesti ossessivi, ripetitivi, senza uno scopo preciso. La condizione dei pazienti affetti da demenza è valutata clinicamente tramite apposite scale e le informazioni relative al comportamento vengono raccolte intervistando chi se ne occupa, come familiari, il personale infermieristico o il medico curante. Spesso queste valutazioni si rivelano inaccurate, possono essere fortemente influenzate da considerazioni soggettive, e sono dispendiose in termini di tempo. Si ha quindi l'esigenza di disporre di metodiche oggettive per valutare il comportamento motorio dei pazienti e le sue alterazioni patologiche; i sensori inerziali indossabili potrebbero costituire una valida soluzione, per questo scopo. L'obiettivo principale della presente attività di tesi è stato definire e implementare un software per una valutazione oggettiva, basata su sensori, del pattern motorio circadiano, in pazienti affetti da demenza ricoverati in un'unità di terapia a lungo termine, che potrebbe evidenziare differenze nei sintomi della malattia che interessano il comportamento motorio, come descritto in ambito clinico. Lo scopo secondario è stato quello di verificare i cambiamenti motori pre- e post-intervento in un sottogruppo di pazienti, a seguito della somministrazione di un programma sperimentale di intervento basato su esercizi fisici. --------------- [ENG]Dementia involves deterioration, often progressive, of a person's cognitive status. Those who suffer from dementia, present alterations in cognitive and motor behavior, for example performing obsessive and repetitive gestures, without a purpose. The condition of patients suffering from dementia is clinically assessed by means of specific scales and information relating to the behavior are collected by interviewing caregivers, such as the family, nurses, or the doctor. Often it turns out that these are inaccurate assessments that may be heavily influenced by subjective evaluations and are costly in terms of time. Therefore, there is the need for objective methods to assess the patients' motor behavior and the pathological changes; wearable inertial sensors may represent a viable option, so this aim. The main objective of this thesis project was to define and implement a software for a sensor-based assessment of the circadian motor pattern in patients suffering from dementia, hospitalized in a long-term care unit, which could highlight differences in the disease symptoms affecting the motor behavior, as described in the clinical setting. The secondary objective was to verify pre- and post-intervention changes in the motor patterns of a subgroup of patients, following the administration of an experimental program of intervention based on physical exercises.