Assessment of PCR in the detection of Leishmania spp in experimentally infected individual phlebotomine sandflies (Diptera: Psychodidae: Phlebotominae)

DNA amplification by the polymerase chain reaction (PCR) was applied in the investigation of the presence of Leishmania (Kinetoplastida: Trypanosomatidae) parasites in single phlebotomine sandflies. Three phlebotomine/parasite pairs were used: Lutzomyia longipalpis/Leishmania chagasi, Lutzomyia migonei/Leishmania amazonensis and Lutzomyia migonei/Leishmania braziliensis, all of them incriminated in the transmission of visceral or cutaneous leishmaniasis. DNA extraction was performed with whole insects, with no need of previous digestive tract dissection or pooling specimens. The presence of either mouse blood in the digestive tract of the sandflies or the digestive tract itself did not interfere in the PCR. Infection by as few as 10 Leishmania sp. per individual were sufficient for DNA amplification with genus-specific primers. Using primers for L. braziliensis and L. mexicana complexes, respectively, it was possible to discriminate between L. braziliensis and L. amazonensis in experimentally infected vectors (L. migonei).

Prometeu: Desenho e desenvolvimento da plataforma digital. Concepção e produção dos conteúdos digitais.

Prometeu, é uma peça de teatro multimédia inspirada no teatro de sombras indonésio, Wayang Kulit.O projecto Prometeu apresenta uma abordagem inovadora misturando a linguagem do teatro de sombras com a linguagem cinematográfica tendo a multimédia como elemento agregador. Um espectáculo de silhuetas/marionetas que promove o cruzamento entre o tradicional e a modernidade, o analógico e o digital, o performativo e o autómato, ou procedural. Uma visão ampla da tecnologia ao serviço da dramaturgia explorando a harmonia entre diferentes meios. Este projecto tem vindo a ser apresentado ao público, desde 2009, em locais e eventos distintos. Festivais de música, festivais de cinema, e festivais de teatro em Portugal e Espanha, são exemplos da abrangência deste espectáculo tendo sido já reconhecido com um prémio. Com o presente trabalho pretende-­‐se demonstrar as competências profissionais a que Luís Leite se apresenta como candidato a especialista. Competências na área das artes da imagem, particularmente na área da multimédia. Luís Leite foi convidado pelo Teatro de Formas Animadas e pela Casa da Música para desenhar e desenvolver uma solução multimédia que respondesse aos complexos requisitos do projecto, bem como para a concepção e produção de conteúdos de animação e pós-produção digital. Competências importantes para as unidades curriculares de animação e multimédia que lecciona.

Preservação digital nas médias e grandes empresas portuguesas: estudo de caso e recomendação de boas práticas

Este estudo tem como objetivo analisar o estado atual da preservação digital no tecido empresarial português, particularmente nas empresas de média e grande dimensão. Para tal, expôs-se o referencial teórico da preservação digital, necessário para uma melhor compreensão desta temática, nomeadamente através da apresentação de definições de diversos autores, de estratégias e boas práticas, e de algumas das iniciativas internacionais mais relevantes. De seguida apresentou-se a importância que a informação tem para uma organização, sobretudo aquela informação criada no âmbito laboral. Realizou-se, também, uma caracterização dos vários tipos de empresas, de acordo com diretivas europeias, e efetuou-se uma breve análise das empresas portuguesas. Para atingir esse objetivo elaborou-se um inquérito, por questionário, fazendo- se uso de uma plataforma de questionários online. Este foi remetido a 1.358 médias e grandes empresas que atuam em Portugal, com o objetivo de compreender – entre outros – a importância dada, pelas empresas, ao material existente em formato digital, quais as tecnologias responsáveis pelo seu armazenamento e quais os cuidados realizados para assegurar o seu acesso por longos períodos de tempo. Ao analisar os resultados, e pese embora a baixa taxa de resposta ao questionário, foi possível concluir que quase todas as empresas participantes já procuram substituir os documentos em papel por documentos em formato digital e que se preocupam em manter o acesso prolongado à informação. Não obstante, dois terços dos participantes conhecem o conceito de preservação digital e apenas 44% afirmam aplicar estratégias desse tipo na sua empresa. Apesar de cerca de 70% dos participantes reconhecerem a importância do uso de formatos abertos para a preservação digital, na realidade ainda se verifica uma utilização massiva de formatos proprietários ao nível dos documentos de texto e das folhas de cálculo. Para além deste contributo analítico sobre o estado da preservação digital no tecido empresarial português, prepararam-se alguns documentos que visam auxiliar a elaboração de um plano de preservação digital e, consequentemente, permitir a sustentabilidade a longo prazo da informação digital.

Diagnosis of abdominal angiostrongyliasis by PCR from sera of patients

Abdominal angiostrongyliasis is a zoonotic infection caused by an intra-vascular nematode parasitic of wild rodents, Angiostrongylus costaricensis. No parasitological diagnosis is currently available and immunodiagnosis presents several drawbacks. Primers constructed based on a congeneric species, A. cantonensis, were able to amplify a 232 bp fragment from serum samples of 3 patients with histopathological diagnosis. Extraction was better performed with DNAzol and the specificity of the primers was confirmed by Southern blot. This disease has been diagnosed with frequency in south of Brazil, thus, this method appears like the important and unpublished alternative to improve diagnostic of disease.

"Multiplex PCR" identification of the atypical and monophasic Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- in São Paulo State, Brazil: frequency and antibiotic resistance patterns

Salmonella spp. are the etiologic agents of salmonellosis, a worldwide spread zoonoses causing foodborne outbreaks and clinical diseases. By serological identification, Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- accounted for 8.8% of human and 1.6% of nonhuman Salmonella strains isolated in São Paulo State, during 1991-2000. A total of 28.6% of them amplified a fragment corresponding to H:1,2 (flagellar phase two) through PCR analysis and were further assigned as S. Typhimurium. Antimicrobial resistance was detected in 36.3% of the 369 PCR-negative strains tested, including the multiresistance to ampicillin, chloramphenicol, sulfonamides, tetracycline, and streptomycin.

PCR - based diagnosis to evaluate the performance of malaria reference centers

Although the Giemsa-stained thick blood smear (GTS) remains the gold standard for the diagnosis of malaria, molecular methods are more sensitive and specific to detect parasites and can be used at reference centers to evaluate the performance of microscopy. The description of the Plasmodium falciparum, P. vivax, P. malariae and P. ovale ssrRNA gene sequences allowed the development of a polymerase chain reaction (PCR) that had been used to differentiate the four species. The objective of this study was to determine Plasmodium species through PCR in 190 positive smears from patients in order to verify the quality of diagnosis at SUCEN's Malaria Laboratory. Considering only the 131 positive results in both techniques, GTS detected 4.6% of mixed and 3.1% of P. malariae infections whereas PCR identified 19.1% and 13.8%, respectively.

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.

Single step polymerase chain reaction (PCR) for the diagnosis of the Leishmania (Viannia) subgenus

In Brazil, the main etiologic agent of Leishmaniasis that frequently presents with mucosal involvement belongs to the Viannia subgenus. The therapeutic conduct in this disease depends on the parasitological diagnosis, and classical methods are restricted in identifying the agent. In this paper we describe a polymerase chain reaction (PCR), which uses primers designed from mini-exons repetitive sequences. The PCR amplifies a 177bp fragment that can distinguish (Viannia) from (Leishmania) subgenus. This test could be a useful diagnostic tool.

Detection of EBV-DNA in serum samples of an immunosuppressed child during a three years follow-up: association of clinical and PCR data with active infection

Twenty-four whole blood and serum samples were drawn from an eight year-old heart transplant child during a 36 months follow-up. EBV serology was positive for VCA-IgM and IgG, and negative for EBNA-IgG at the age of five years old when the child presented with signs and symptoms suggestive of acute infectious mononucleosis. After 14 months, serological parameters were: positive VCA-IgG, EBNA-IgG and negative VCA-IgM. This serological pattern has been maintained since then even during episodes suggestive of EBV reactivation. PCR amplified a specific DNA fragment from the EBV gp220 (detection limit of 100 viral copies). All twenty-four whole blood samples yielded positive results by PCR, while 12 out of 24 serum samples were positive. We aimed at analyzing whether detection of EBV-DNA in serum samples by PCR was associated with overt disease as stated by the need of antiviral treatment and hospitalization. Statistical analysis showed agreement between the two parameters evidenced by the Kappa test (value 0.750; p < 0.001). We concluded that detection of EBV-DNA in serum samples of immunosuppressed patients might be used as a laboratory marker of active EBV disease when a Real-Time PCR or another quantitative method is not available.

Students’ reading behavior: digital vs. print preferences in portuguese context

The aim of this paper is to present the main Portuguese results from a multi-national study on reading format preferences and behaviors from undergraduate students from Polytechnic Institute of Porto (Portugal). For this purpose we apply an adaptation of the Academic Reading Questionnaire previously created by Mizrachi (2014). This survey instrument has 14 Likert-style statements regarding the format influence in the students reading behavior, including aspects such as ability to remember, feelings about access convenience, active engagement with the text by highlighting and annotating, and ability to review and concentrate on the text. The importance of the language and dimension of the text to determine the preference format is also inquired. Students are also asked about the electronic device they use to read digital documents. Finally, some demographic and academic data were gathered. The analysis of the results will be contextualized on a review of the literature concerning youngsters reading format preferences. The format (digital or print) in which a text is displayed and read can impact comprehension, which is an important information literacy skill. This is a quite relevant issue for class readings in academic context because it impacts learning. On the other hand, students preferences on reading formats will influence the use of library services. However, literature is not unanimous on this subject. Woody, Daniel and Baker (2010) concluded that the experience of reading is not the same in electronic or print context and that students prefer print books than e-books. This thesis is reinforced by Ji, Michaels and Waterman (2014) which report that among 101 undergraduates the large majority self-reported to read and learn more when they use printed format despite the fact that they prefer electronically supplied readings instead of those supplied in printed form. On the other side, Rockinson-Szapkiw, et al (2013) conducted a study were they demonstrate that e-textbook is as effective for learning as the traditional textbook and that students who choose e-textbook had significantly higher perceived learning than students who chose to use print textbooks.

Design of Digital Fractional-Order Integrators and Differentiators by Least Squares

First IFAC Workshop on Fractional Differentiation and Its Application - 19-21 July 2004, Enseirb, Bordeaux, France - FDA'04

New Concepts Towards the Synthesis of Digital Circuits Through Genetic Algorithms

This paper analyses the performance of a Genetic Algorithm using two new concepts, namely a static fitness function including a discontinuity measure and a fractional-order dynamic fitness function, for the synthesis of combinational logic circuits. In both cases, experiments reveal superior results in terms of speed and convergence to achieve a solution.

SI SC MDAC switched current-capacitor multiplying digital-to-analog converter

Dissertação apresentada para a obtenção do grau de Mestre em Engenharia Electrotécnica e de Computadores

A migração para o digital. A luta pela sobrevivência dos média tradicionais, num ambiente Web hostil. O caso da Agência de Notícias Lusa.

Dissertação e Trabalho de Projecto apresentados para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Novos Media e Práticas Web

Fecal specimens preparation methods for PCR diagnosis of human taeniosis

Sample preparation and DNA extraction protocols for DNA amplification by PCR, which can be applied in human fecal samples for taeniasis diagnosis, are described. DNA extracted from fecal specimens with phenol/chloroform/isoamilic alcohol and DNAzol® reagent had to be first purified to generate fragments of 170 pb and 600 pb by HDP2-PCR. This purification step was not necessary with the use of QIAmp DNA stool mini kit®. Best DNA extraction results were achieved after eggs disruption with glass beads, either with phenol/chloroform/isoamilic alcohol, DNAzol® reagent or QIAmp DNA stool mini kit®.