Neurocryptococcosis: diagnosis by PCR method

Cryptococcus neoformans detection was optimized using PCR technique with the objective of application in the clinical laboratory diagnosis. The amplification area was ITS and 5,6S which encodes the ribosomal RNA (rRNA). A total of 72 cerebrospinal fluid (CSF) samples were used, obtained from cases with and without AIDS. The patients had cryptococcal meningitis (n = 56) and meningitis caused by other agents (n = 16). The results demonstrated that PCR test had the highest sensitivity rates, superior to culture (85.7%) and to India ink test (76.8%). PCR was found to be sensitive in detecting 1 cell/mL and highly specific since it did not amplify other fungal DNA. The comparative analysis of the methods showed that PCR is more sensitive and specific and is applicable as an important laboratorial resource for neurocryptococcosis diagnosis.

Fractional State Space Analysis of Economy Systems

This paper examines modern economic growth according to the multidimensional scaling (MDS) method and state space portrait (SSP) analysis. Electing GDP per capita as the main indicator for economic growth and prosperity, the long-run perspective from 1870 to 2010 identifies the main similarities among 34 world partners’ modern economic growth and exemplifies the historical waving mechanics of the largest world economy, the USA. MDS reveals two main clusters among the European countries and their old offshore territories, and SSP identifies the Great Depression as a mild challenge to the American global performance, when compared to the Second World War and the 2008 crisis.

Uma perspectiva foucaudiana na auditoria à avaliação de desempenho na administração pública

Motivation: Auditing is not merely a collection of technical tasks but also a programmatic idea circulating in organizational environment, an idea which promises a certain style of control and organizational transparency (Power, 1998, p. 122) Performance appraisal within public organization aims to promote this organizational transparency and promote learning and improvement process both for employees and for the organization. However, we suggest that behind its clear intentions, there are some other goals tied to performance appraisal that could be seen as components of a discipline and surveillance systems to make the employee “knowable, calculable and administrative object” (Miller and Rose, 1990, p. 5). Objective: In Portuguese public organizations, performance appraisal follows the SIADAP (Performance Appraisal Systems for Public Administration). The objective of this study is to capture whatever employees of public organizations (appraisers and appraisee) perceived the performance appraisal system (SIADAP) as an appraisal model that promotes equity, learning and improvement or just as an instrument of control to which they feel dominated and watched over. Method: We developed an in-depth qualitative case study using semi-structured interviews with appraisers and their subordinates in the administrative department of a university institute of Medicine. The discourse of the participants was theoretically analyzed based on Foucauldian framework. Prior to qualitative data collection, we collected quantitative data, with a questionnaire, to measure the (un)satisfaction of employees with the all appraisal system. Findings: Although some key points of Foucault perspective were identified, its framework revealed some limitations to capture the all complexity of performance appraisal. Qualitative data revealed a significant tendency in discourses of appraisers and their subordinates considering SIADAP as an instrument that’s aims to introduced political rationalities and limits to the employer’s promotions within their careers. Contribution: This study brings a critical perspectives and new insights about performance appraisals in Portuguese’s public administrations. It is original contribution to management of human recourses in public administration and primary to audit of performance appraisal systems.

A low-cost method to test cytotoxic effects of Crotalus vegrandis (Serpentes: Viperidae) venom on kidney cell cultures

The pathogenesis of the renal lesion upon envenomation by snakebite has been related to myolysis, hemolysis, hypotension and/or direct venom nephrotoxicity caused by the venom. Both primary and continuous cell culture systems provide an in vitro alternative for quantitative evaluation of the toxicity of snake venoms. Crude Crotalus vegrandis venom was fractionated by molecular exclusion chromatography. The toxicity of C. vegrandis crude venom, hemorrhagic, and neurotoxic fractions were evaluated on mouse primary renal cells and a continuous cell line of Vero cells maintained in vitro. Cells were isolated from murine renal cortex and were grown in 96 well plates with Dulbecco's Modified Essential Medium (DMEM) and challenged with crude and venom fractions. The murine renal cortex cells exhibited epithelial morphology and the majority showed smooth muscle actin determined by immune-staining. The cytotoxicity was evaluated by the tetrazolium colorimetric method. Cell viability was less for crude venom, followed by the hemorrhagic and neurotoxic fractions with a CT50 of 4.93, 18.41 and 50.22 µg/mL, respectively. The Vero cell cultures seemed to be more sensitive with a CT50 of 2.9 and 1.4 µg/mL for crude venom and the hemorrhagic peak, respectively. The results of this study show the potential of using cell culture system to evaluate venom toxicity.

Scalable and Efficient Configuration of TimeDivision Multiplexed Resources

Consumer-electronics systems are becoming increasingly complex as the number of integrated applications is growing. Some of these applications have real-time requirements, while other non-real-time applications only require good average performance. For cost-efficient design, contemporary platforms feature an increasing number of cores that share resources, such as memories and interconnects. However, resource sharing causes contention that must be resolved by a resource arbiter, such as Time-Division Multiplexing. A key challenge is to configure this arbiter to satisfy the bandwidth and latency requirements of the real-time applications, while maximizing the slack capacity to improve performance of their non-real-time counterparts. As this configuration problem is NP-hard, a sophisticated automated configuration method is required to avoid negatively impacting design time. The main contributions of this article are: 1) An optimal approach that takes an existing integer linear programming (ILP) model addressing the problem and wraps it in a branch-and-price framework to improve scalability. 2) A faster heuristic algorithm that typically provides near-optimal solutions. 3) An experimental evaluation that quantitatively compares the branch-and-price approach to the previously formulated ILP model and the proposed heuristic. 4) A case study of an HD video and graphics processing system that demonstrates the practical applicability of the approach.

Calibration Method for Underwater Visual Ground-Truth System

This work presents an automatic calibration method for a vision based external underwater ground-truth positioning system. These systems are a relevant tool in benchmarking and assessing the quality of research in underwater robotics applications. A stereo vision system can in suitable environments such as test tanks or in clear water conditions provide accurate position with low cost and flexible operation. In this work we present a two step extrinsic camera parameter calibration procedure in order to reduce the setup time and provide accurate results. The proposed method uses a planar homography decomposition in order to determine the relative camera poses and the determination of vanishing points of detected lines in the image to obtain the global pose of the stereo rig in the reference frame. This method was applied to our external vision based ground-truth at the INESC TEC/Robotics test tank. Results are presented in comparison with an precise calibration performed using points obtained from an accurate 3D LIDAR modelling of the environment.

Previsão do Diagrama de Carga de Subestações da REN utilizando Redes Neuronais

A presente dissertação apresenta o estudo de previsão do diagrama de carga de subestações da Rede Elétrica Nacional (REN) utilizando redes neuronais, com o intuito de verificar a viabilidade do método utilizado, em estudos futuros. Atualmente, a energia elétrica é um bem essencial e desempenha um papel fundamental, tanto a nível económico do país, como a nível de conforto e satisfação individual. Com o desenvolvimento do setor elétrico e o aumento dos produtores torna-se importante a realização da previsão de diagramas de carga, contribuindo para a eficiência das empresas. Esta dissertação tem como objetivo a utilização do modelo das redes neuronais artificiais (RNA) para criar uma rede capaz de realizar a previsão de diagramas de carga, com a finalidade de oferecer a possibilidade de redução de custos e gastos, e a melhoria de qualidade e fiabilidade. Ao longo do trabalho são utilizados dados da carga (em MW), obtidos da REN, da subestação da Prelada e dados como a temperatura, humidade, vento e luminosidade, entre outros. Os dados foram devidamente tratados com a ajuda do software Excel. Com o software MATLAB são realizados treinos com redes neuronais, através da ferramenta Neural Network Fitting Tool, com o objetivo de obter uma rede que forneça os melhores resultados e posteriormente utiliza-la na previsão de novos dados. No processo de previsão, utilizando dados reais das subestações da Prelada e Ermesinde referentes a Março de 2015, comprova-se que com a utilização de RNA é possível obter dados de previsão credíveis, apesar de não ser uma previsão exata. Deste modo, no que diz respeito à previsão de diagramas de carga, as RNA são um bom método a utilizar, uma vez que fornecem, à parte interessada, uma boa previsão do consumo e comportamento das cargas elétricas. Com a finalização deste estudo os resultados obtidos são no mínimo satisfatórios. Consegue-se alcançar através das RNA resultados próximos aos valores que eram esperados, embora não exatamente iguais devido à existência de uma margem de erro na aprendizagem da rede neuronal.

Replacement of HIV p24 Ag test by a multiplex RT-PCR method for primary screening of blood donors

Nucleic Acid Testing (NAT) as a tool for primary screening of blood donors became a reality in the end of the 1990 decade. We report here the development of an "in-house" RT-PCR method that allows the simultaneous (multiplex) detection of HCV and HIV-RNA in addition to an artificial RNA employed as an external control. This method detects all HIV group M subtypes, plus group N and O, with a detection threshold of 500 IU/mL. After validation, the method replaced p24 Ag testing, in use for blood donation screening since 1996 at our services. From July 2001 to February 2006, 102,469 donations were tested and 41 (0.04%) were found HIV-RNA reactive. One NAT-only reactive donation (antibody non-reactive) was observed, with subsequent seroconversion of the implied donor, giving a yield of 1:102,469. This rate is in contrast to the international experience that reports a detection of approximately 1:600,000 - 1:3,100,000 of isolated HIV-RNA donations.

Primary screening of blood donors by nat testing for HCV-RNA: development of an "in-house" method and results

An "in-house" RT-PCR method was developed that allows the simultaneous detection of the RNA of the Hepatitis C Virus (HCV) and an artificial RNA employed as an external control. Samples were analyzed in pools of 6-12 donations, each donation included in two pools, one horizontal and one vertical, permitting the immediate identification of a reactive donation, obviating the need for pool dismembering. The whole process took 6-8 hours per day and results were issued in parallel to serology. The method was shown to detect all six HCV genotypes and a sensitivity of 500 IU/mL was achieved (95% hit rate). Until July 2005, 139,678 donations were tested and 315 (0.23%) were found reactive for HCV-RNA. Except for five false-positives, all 310 presented the corresponding antibody as well, so the yield of NAT-only donations was zero, presenting a specificity of 99.83%. Detection of a window period donation, in the population studied, will probably demand testing of a larger number of donations. International experience is showing a rate of 1:200,000 - 1:500,000 of isolated HCV-RNA reactive donations.

A potentiometric magnetic immunoassay for rapid detection of Salmonella typhimurium

Potentiometric detection with homemade polymeric membrane microelectrodes was coupled to a magnetic sandwich immunoassay for Salmonella typhimurium determination. Cadmium and sodium ion selective electrodes were used respectively as indicator and pseudo-reference electrodes and were prepared in pipette tips to allow potentiometric measurements in microliter sample volumes. In the proposed method, the concentration of S. typhimurium was proportional to the amount of cadmium released upon dissolution of a CdS nanoparticle labeled to the secondary detection antibody. The limit of detection was 2 cells per 100 μL. The immunomagnetic assay with potentiometric detection is suitable for sensitive and rapid (average total time per assay of 75 minutes) detection of S. typhimurium in milk samples. The proposed method is easy to perform, safe, sensitive, and low cost and has potential for in situ analysis.

Validation of QuEChERS method for organochlorine pesticides analysis in tamarind (Tamarindus indica) products: Peel, fruit and commercial pulp

In this study a citrate-buffered version of QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method for determination of 14 organochlorine pesticides (OCPs) residues in tamarind peel, fruit and commercial pulp was optimized using gas chromatography (GC) coupled with electron-capture detector (ECD) and confirmation by GC tandem mass spectrometry (GC–MS/MS). Five procedures were tested based on the original QuEChERS method. The best one was achieved with increased time in ultrasonic bath. For the extract clean-up, primary secondary amine (PSA), octadecyl-bonded silica (C18) and magnesium sulphate (MgSO4) were used as sorbents for tamarind fruit and commercial pulp and for peel was also added graphitized carbon black (GCB). The samples mass was optimized according to the best recoveries (1.0 g for peel and fruit; 0.5 g for pulp). The method results showed the matrix-matched calibration curve linearity was r2 > 0.99 for all target analytes in all samples. The overall average recoveries (spiked at 20, 40 and 60 μg kg−1) have been considered satisfactory presenting values between 70 and 115% with RSD of 2–15 % (n = 3) for all analytes, with the exception of HCB (in peel sample). The ranges of limits of detection (LOD) and quantification (LOQ) for OCPs were for peel (LOD: 8.0–21 μg kg−1; LOQ: 27–98 μg kg−1); for fruit (LOD: 4–10 μg kg−1; LOQ: 15–49 μg kg−1) and for commercial pulp (LOD: 2–5 μg kg−1; LOQ: 7–27 μg kg−1). The method was successfully applied in tamarind samples being considered a rapid, sensitive and reliable procedure.

Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates

Thirty Candida albicans isolated from oral candidosis patients and 30 C. albicans isolated from control individuals were studied. In vitro susceptibility tests were performed for amphotericin B, fluconazole, 5-flucytosine and itraconazole through the Clinical and Laboratorial Standards Institute (CLSI) reference method and E test system. The results obtained were analyzed and compared. MIC values were similar for the strains isolated from oral candidosis patients and control individuals. The agreement rate for the two methods was 66.67% for amphotericin B, 53.33% for fluconazole, 65% for flucytosine and 45% for itraconazole. According to our data, E test method could be an alternative to trial routine susceptibility testing due to its simplicity. However, it can not be considered a substitute for the CLSI reference method.

An integration bridge for heterogeneous e-service environments

Dissertação para obtenção do Grau de Mestre em Engenharia Electrotécnica e de Computadores

Optimization of the Tartrate-Resistant Acid Phosphatase Detection by Histochemical Method

According to the new KDIGO (Kidney Disease Improving Global Outcomes) guidelines, the term of renal osteodystrophy, should be used exclusively in reference to the invasive diagnosis of bone abnormalities. Due to the low sensitivity and specificity of biochemical serum markers of bone remodelling,the performance of bone biopsies is highly stimulated in dialysis patients and after kidney transplantation. The tartrate-resistant acid phosphatase (TRACP) is an iso-enzyme of the group of acid phosphatases, which is highly expressed by activated osteoclasts and macrophages. TRACP in osteoclasts is in intracytoplasmic vesicles that transport the products of bone matrix degradation. Being present in activated osteoclasts, the identification of this enzyme by histochemistry in undecalcified bone biopsies is an excellent method to quantify the resorption of bone. Since it is an enzymatic histochemical method for a thermolabile enzyme, the temperature at which it is performed is particularly relevant. This study aimed to determine the optimal temperature for identification of TRACP in activated osteoclasts in undecalcified bone biopsies embedded in methylmethacrylate. We selected 10 cases of undecalcified bone biopsies from hemodialysis patients with the diagnosis of secondary hyperparathyroidism. Sections of 5 μm were stained to identify TRACP at different incubation temperatures (37º, 45º, 60º, 70º and 80ºC) for 30 minutes. Activated osteoclasts stained red and trabecular bone (mineralized bone) was contrasted with toluidine blue. This approach also increased the visibility of the trabecular bone resorption areas (Howship lacunae). Unlike what is suggested in the literature and in several international protocols, we found that the best results were obtained with temperatures between 60ºC and 70ºC. For technical reasons and according to the results of the present study, we recommended that, for an incubation time of 30 minutes, the reaction should be carried out at 60ºC. As active osteoclasts are usually scarce in a bone section, the standardization of the histochemistry method is of great relevance, to optimize the identification of these cells and increase the accuracy of the histomosphometric results. Our results, allowing an increase in osteoclasts contrast, also support the use of semi-automatic histomorphometric measurements.