In vitro bioassay investigations of the endocrine disrupting potential of steviol glycosides and their metabolite steviol, components of the natural sweetener Stevia

The food industry is moving towards the use of natural sweeteners such as those produced by Stevia rebaudiana due to the number of health and safety concerns surrounding artificial sweeteners. Despite the fact that these sweeteners are natural; they cannot be assumed safe. Steviol glycosides have a steroidal structure and therefore may have the potential to act as an endocrine disruptor in the body. Reporter gene assays (RGAs), H295R steroidogenesis assay and Ca(2+) fluorimetry based assays using human sperm cells have been used to assess the endocrine disrupting potential of two steviol glycosides: stevioside and rebaudioside A, and their metabolite steviol. A decrease in transcriptional activity of the progestagen receptor was seen following treatment with 25,000 ng/ml steviol in the presence of progesterone (157 ng/ml) resulting in a 31% decrease in progestagen response (p=<0.01). At the level of steroidogenesis, the metabolite steviol (500-25,000 ng/ml) increased progesterone production significantly by 2.3 fold when exposed to 10,000 ng/ml (p=<0.05) and 5 fold when exposed to 25,000 ng/ml (p=<0.001). Additionally, steviol was found to induce an agonistic response on CatSper, a progesterone receptor of sperm, causing a rapid influx of Ca(2+). The response was fully inhibited using a specific CatSper inhibitor. These findings highlight the potential for steviol to act as a potential endocrine disruptor.

DACA: architecture to implement dynamic access control mechanisms on business tier components

Access control is a software engineering challenge in database applications. Currently, there is no satisfactory solution to dynamically implement evolving fine-grained access control mechanisms (FGACM) on business tiers of relational database applications. To tackle this access control gap, we propose an architecture, herein referred to as Dynamic Access Control Architecture (DACA). DACA allows FGACM to be dynamically built and updated at runtime in accordance with the established fine-grained access control policies (FGACP). DACA explores and makes use of Call Level Interfaces (CLI) features to implement FGACM on business tiers. Among the features, we emphasize their performance and their multiple access modes to data residing on relational databases. The different access modes of CLI are wrapped by typed objects driven by FGACM, which are built and updated at runtime. Programmers prescind of traditional access modes of CLI and start using the ones dynamically implemented and updated. DACA comprises three main components: Policy Server (repository of metadata for FGACM), Dynamic Access Control Component (DACC) (business tier component responsible for implementing FGACM) and Policy Manager (broker between DACC and Policy Server). Unlike current approaches, DACA is not dependent on any particular access control model or on any access control policy, this way promoting its applicability to a wide range of different situations. In order to validate DACA, a solution based on Java, Java Database Connectivity (JDBC) and SQL Server was devised and implemented. Two evaluations were carried out. The first one evaluates DACA capability to implement and update FGACM dynamically, at runtime, and, the second one assesses DACA performance against a standard use of JDBC without any FGACM. The collected results show that DACA is an effective approach for implementing evolving FGACM on business tiers based on Call Level Interfaces, in this case JDBC.

Fiber-optic components for optical communicatios and sensing

Nos últimos anos, a Optoelectrónica tem sido estabelecida como um campo de investigação capaz de conduzir a novas soluções tecnológicas. As conquistas abundantes no campo da óptica e lasers, bem como em comunicações ópticas têm sido de grande importância e desencadearam uma série de inovações. Entre o grande número de componentes ópticos existentes, os componentes baseados em fibra óptica são principalmente relevantes devido à sua simplicidade e à elevada de transporte de dados da fibra óptica. Neste trabalho foi focado um destes componentes ópticos: as redes de difracção em fibra óptica, as quais têm propriedades ópticas de processamento únicas. Esta classe de componentes ópticos é extremamente atraente para o desenvolvimento de dispositivos de comunicações ópticas e sensores. O trabalho começou com uma análise teórica aplicada a redes em fibra e foram focados os métodos de fabricação de redes em fibra mais utilizados. A inscrição de redes em fibra também foi abordado neste trabalho, onde um sistema de inscrição automatizada foi implementada para a fibra óptica de sílica, e os resultados experimentais mostraram uma boa aproximação ao estudo de simulação. Também foi desenvolvido um sistema de inscrição de redes de Bragg em fibra óptica de plástico. Foi apresentado um estudo detalhado da modulação acústico-óptica em redes em fibra óptica de sílica e de plástico. Por meio de uma análise detalhada dos modos de excitação mecânica aplicadas ao modulador acústico-óptico, destacou-se que dois modos predominantes de excitação acústica pode ser estabelecidos na fibra óptica, dependendo da frequência acústica aplicada. Através dessa caracterização, foi possível desenvolver novas aplicações para comunicações ópticas. Estudos e implementação de diferentes dispositivos baseados em redes em fibra foram realizados, usando o efeito acústico-óptico e o processo de regeneração em fibra óptica para várias aplicações tais como rápido multiplexador óptico add-drop, atraso de grupo sintonizável de redes de Bragg, redes de Bragg com descolamento de fase sintonizáveis, método para a inscrição de redes de Bragg com perfis complexos, filtro sintonizável para equalização de ganho e filtros ópticos notch ajustáveis.

Geofencing Components and Existing Models

This paper describes the various Geofencing Components and Existing Models in terms of their Information Security Control Attribute Profiles. The profiles will dictate the security attributes that should accompany each and every Geofencing Model used for Wi-Fi network security control in an organization, thus minimizing the likelihood of malfunctioning security controls. Although it is up to an organization to investigate the best way of implementing information security for itself, by looking at the related models that have been used in the past this paper will present models commonly used to implement information security controls in the organizations. Our findings will highlight the strengths and weaknesses of the various models and present what our experiment and prototype consider as a robust Geofencing Security Model for securing Wi-Fi Networks

Biological membranes : biophysical properties and aquaporin function

Tese de doutoramento, Farmácia (Bioquímica), Universidade de Lisboa, Faculdade de Farmácia, 2014

Forest soil short term response to prescribed fire: an approach by Principal Components Analysis

In the current context of serious climate changes, where the increase of the frequency of some extreme events occurrence can enhance the rate of periods prone to high intensity forest fires, the National Forest Authority often implements, in several Portuguese forest areas, a regular set of measures in order to control the amount of fuel mass availability (PNDFCI, 2008). In the present work we’ll present a preliminary analysis concerning the assessment of the consequences given by the implementation of prescribed fire measures to control the amount of fuel mass in soil recovery, in particular in terms of its water retention capacity, its organic matter content, pH and content of iron. This work is included in a larger study (Meira-Castro, 2009(a); Meira-Castro, 2009(b)). According to the established praxis on the data collection, embodied in multidimensional matrices of n columns (variables in analysis) by p lines (sampled areas at different depths), and also considering the quantitative data nature present in this study, we’ve chosen a methodological approach that considers the multivariate statistical analysis, in particular, the Principal Component Analysis (PCA ) (Góis, 2004). The experiments were carried out in a soil cover over a natural site of Andaluzitic schist, in Gramelas, Caminha, NW Portugal, who was able to maintain itself intact from prescribed burnings from four years and was submit to prescribed fire in March 2008. The soils samples were collected from five different plots at six different time periods. The methodological option that was adopted have allowed us to identify the most relevant relational structures inside the n variables, the p samples and in two sets at the same time (Garcia-Pereira, 1990). Consequently, and in addition to the traditional outputs produced from the PCA, we have analyzed the influence of both sampling depths and geomorphological environments in the behavior of all variables involved.

Effect of active components from Garlic on AGEs formation

Advanced glycation end-products are Maillard reaction products that are found in thermal processed food. This compounds are often referred as unhealthy for human diet, namely because of their capacity to form amino-acid dimers. There is a broad range of answers to get about how these products are formed, how they interact with the organism and how these reactions can be inhibited to prevent the referred effects. Some compounds from garlic are thought to be able to inhibit these reactions. This study using spectrophotometric, High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS) and Fourier transformed infrared spectroscopy (FTIR) analysis, helps to understand better not only not only the effect of some compounds obtained from garlic, diallyl sulfide (DAS), diallyl disulfide (DADS) and diallyl trisulfide (DATS), on these AGEs production reaction, but also helped to understand better the reaction itself.

The effect of method, standard and sample components on the total antioxidant capacity of commercial waters assessed by optical conventional assays

The total antioxidant capacity (TAC) of 28 flavoured water samples was assessed by ferric reducing antioxidant potential (FRAP), oxygen radical absorbance capacity (ORAC), trolox equivalent antioxidant capacity (TEAC) and total reactive antioxidant potential (TRAP) methods. It was observed that flavoured waters had higher antioxidant activity than the corresponding natural ones. The observed differences were attributed to flavours, juice and vitamins. Generally, higher TAC contents were obtained on lemon waters and lower values on guava and raspberry flavoured waters. Lower and higher TACs were obtained by TRAP and ORAC method, respectively. Statistical analysis suggested that vitamins and flavours increased the antioxidant content of the commercial waters.

Pozzolanic components in lime mortars: correlating behaviour, composition and microstructure.

Int. J. for Restoration of Buildings and Monuments, vol.11, nº 2 (2005), p.111-118

A mathematical model to describe fat oxidation kinetics during graded exercise.

PURPOSE: The purpose of this study was to develop a mathematical model (sine model, SIN) to describe fat oxidation kinetics as a function of the relative exercise intensity [% of maximal oxygen uptake (%VO2max)] during graded exercise and to determine the exercise intensity (Fatmax) that elicits maximal fat oxidation (MFO) and the intensity at which the fat oxidation becomes negligible (Fatmin). This model included three independent variables (dilatation, symmetry, and translation) that incorporated primary expected modulations of the curve because of training level or body composition. METHODS: Thirty-two healthy volunteers (17 women and 15 men) performed a graded exercise test on a cycle ergometer, with 3-min stages and 20-W increments. Substrate oxidation rates were determined using indirect calorimetry. SIN was compared with measured values (MV) and with other methods currently used [i.e., the RER method (MRER) and third polynomial curves (P3)]. RESULTS: There was no significant difference in the fitting accuracy between SIN and P3 (P = 0.157), whereas MRER was less precise than SIN (P < 0.001). Fatmax (44 +/- 10% VO2max) and MFO (0.37 +/- 0.16 g x min(-1)) determined using SIN were significantly correlated with MV, P3, and MRER (P < 0.001). The variable of dilatation was correlated with Fatmax, Fatmin, and MFO (r = 0.79, r = 0.67, and r = 0.60, respectively, P < 0.001). CONCLUSIONS: The SIN model presents the same precision as other methods currently used in the determination of Fatmax and MFO but in addition allows calculation of Fatmin. Moreover, the three independent variables are directly related to the main expected modulations of the fat oxidation curve. SIN, therefore, seems to be an appropriate tool in analyzing fat oxidation kinetics obtained during graded exercise.

Extracellular matrix components in peripheral nerve repair: how to affect neural cellular response and nerve regeneration?

Peripheral nerve injury is a serious problem affecting significantly patients' life. Autografts are the "gold standard" used to repair the injury gap, however, only 50% of patients fully recover from the trauma. Artificial conduits are a valid alternative to repairing peripheral nerve. They aim at confining the nerve environment throughout the regeneration process, and providing guidance to axon outgrowth. Biocompatible materials have been carefully designed to reduce inflammation and scar tissue formation, but modifications of the inner lumen are still required in order to optimise the scaffolds. Biomicking the native neural tissue with extracellular matrix fillers or coatings showed great promises in repairing longer gaps and extending cell survival. In addition, extracellular matrix molecules provide a platform to further bind growth factors that can be released in the system over time. Alternatively, conduit fillers can be used for cell transplantation at the injury site, reducing the lag time required for endogenous Schwann cells to proliferate and take part in the regeneration process. This review provides an overview on the importance of extracellular matrix molecules in peripheral nerve repair.

In vitro engineering of human stratified urothelium: analysis of its morphology and function.

PURPOSE: Gastric or intestinal patches, commonly used for reconstructive cystoplasty, may induce severe metabolic complications. The use of bladder tissues reconstructed in vitro could avoid these complications. We compared cellular differentiation and permeability characteristics of human native with in vitro cultured stratified urothelium. MATERIALS AND METHODS: Human stratified urothelium was induced in vitro. Morphology was studied with light and electron microscopy and expression of key cellular proteins was assessed using immunohistochemistry. Permeability coefficients were determined by measuring water, urea, ammonia and proton fluxes across the urothelium. RESULTS: As in native urothelium the stratified urothelial construct consisted of basal membrane and basal, intermediate and superficial cell layers. The apical membrane of superficial cells formed villi and glycocalices, and tight junctions and desmosomes were developed. Immunohistochemistry showed similarities and differences in the expression of cytokeratins, integrin and cellular adhesion proteins. In the cultured urothelium cytokeratin 20 and integrin subunits alpha6 and beta4 were absent, and symplekin was expressed diffusely in all layers. Uroplakins were clearly expressed in the superficial umbrella cells of the urothelial constructs, however, they were also present in intermediate and basal cells. Symplekin and uroplakins were expressed only in the superficial cells of native bladder tissue. The urothelial constructs showed excellent viability, and functionally their permeabilities for water, urea and ammonia were no different from those measured in native human urothelium. Proton permeability was even lower in the constructs compared to that of native urothelium. CONCLUSIONS: Although the in vitro cultured human stratified urothelium did not show complete terminal differentiation of its superficial cells, it retained the same barrier characteristics against the principal urine components. These results indicate that such in vitro cultured urothelium, after being grown on a compliant degradable support or in coculture with smooth muscle cells, is suitable for reconstructive cystoplasty.

Is GABA involved in regulating plant growth and development? /

Rapid and large accumulation of GABA (y-aminobutyric acid) in response to a number of plant stresses has been well documented. But the role(s) of GABA in plants is not well defined. In recent years, the possibility of GABA involvement in regulating plant growth and development has been raised. In the present study, this possibility was examined. First, to rapidly and accurately determine GABA levels in plant tissues, a spectrometric method for GABA determination was developed based on a commercially available enzyme Gabase. Seventy mM LaCb almost completely removed water-soluble pigments from plant tissues which greatly interfere with the absorbance reading at 340nm. Inactivation of GAD (glutamate decarboxylase) by immediately adding methanol to a frozen plant tissue powder was suggested to prevent GABA production during extraction. The recovery of GABA with this method was approximately 100%. Second, the relationship between GABA levels and hypocotyl elongation in soybean seedlings was analyzed using different approaches to regulate in vivo GABA levels and the elongation of hypocotyls. The following major observations were made. (1) Mechanical stimulation by stroking elevated GABA levels and concurrently induced a rapid and significant reduction in hypocotyl elongation. (2) External GABA was demonstrated to penetrate into the hypocotyls using '*C-GABA. Application of external GABA elevated in vivo GABA levels, but failed to inhibit hypocotyl elongation. (3) LaCla and blue light irradiation caused an inhibition in the elongation of dark-grown hypocotyls, whereas GABA levels were not significantly affected. (4) Ca^was suggested to be involved in the signal transduction pathway leading from mechanical stimulation to GABA production, as indicated by the ability of La'* to inhibit GABA production in stimulated hypocotyls. (5) Bicuculline, saclofen and baclofen (agonists and antagonists of GABA receptors in animals) had no effect on hypocotyl elongation. It might indicate that GABA-binding components which are structurally similar to animal GABA receptors and functionally capable of regulating plant growth may not exist in plants. Therefore, the conclusion was drawn that GABA alone is not sufficient to inhibit hypocotyl elongation. Third, chloride influx in isolated Asparagus cells was enhanced by lOmM GABA during a 3 hour incubation, but the effect was not specific for GABA. Chloride efflux was not influenced by GABA. Both influx and efflux of chloride were significantly inhibited by NPPB, a chloride channel blocker. These results suggest that GABA does not influence the activity of plant chloride channels.

Electrophysiological correlates of performance monitoring in middle and late adolescence

The ability to monitor and evaluate the consequences of ongoing behaviors and coordinate behavioral adjustments seems to rely on networks including the anterior cingulate cortex (ACC) and phasic changes in dopamine activity. Activity (and presumably functional maturation) of the ACC may be indirectly measured using the error-related negativity (ERN), an event-related potential (ERP) component that is hypothesized to reflect activity of the automatic response monitoring system. To date, no studies have examined the measurement reliability of the ERN as a trait-like measure of response monitoring, its development in mid- and late- adolescence as well as its relation to risk-taking and empathic ability, two traits linked to dopaminergic and ACC activity. Utilizing a large sample of 15- and 18-year-old males, the present study examined the test-retest reliability of the ERN, age-related changes in the ERN and other components of the ERP associated with error monitoring (the Pe and CRN), and the relations of the error-related ERP components to personality traits of risk propensity and empathy. Results indicated good test-retest reliability of the ERN providing important validation of the ERN as a stable and possibly trait-like electrophysiological correlate of performance monitoring. Ofthe three components, only the ERN was of greater amplitude for the older adolescents suggesting that its ACC network is functionally late to mature, due to either structural or neurochemical changes with age. Finally, the ERN was smaller for those with high risk propensity and low empathy, while other components associated with error monitoring were not, which suggests that poor ACe function may be associated with the desire to engage in risky behaviors and the ERN may be influenced by the extent of individuals' concern with the outcome of events.

Improving the oral health status of functionally independent and dependent seniors residing in long-term care facilities through dental hygiene education /

The purpose of this study was to evaluate the oral health status of residents residing in 2 long-term care facilities and determine if dental hygiene education was required in order to improve their current oral health. The oral health status of 6 independent and 4 dependent individuals residing in 2 long-term care facilities was evaluated. In addition, the current oral health and disease prevention practices employed by 4 caregivers who were responsible for providing oral care to dependent residents in the long-term care facilities were evaluated. Furthermore, an evaluation of the oral care practices of independent residents who were responsible for providing their own care was conducted. Finally, the challenges that caregivers and independent residents faced when performing oral care were determined, and methodological changes were proposed. Using a generic qualitative research methodology, data collection was comprised of semi structured interviews, field observations, and documentation. The oral health status of the residents was reevaluated 3 months later. The findings of this study demonstrated an increase in plaque accumulation, gingival inflammation, and unhealthy gingival tissue colour changes among the residents over the 3-month period. The study revealed that poor oral health among the residents was a result of inadequate oral hygiene care techniques, difficulties accessing oral health care, financial limitations, insufficient care staff, insufficient time for personal care duties, lack of professional development, minimal interprofessional collaboration of health disciplines, and lack of perseverance on the part of the caregivers and residents. Overall, oral health is essential, and maintaining optimal oral health requires increased collaboration and communication between health care providers.