Evaluation of three different DNA extraction methods from blood samples collected in dried filter paper in Plasmodium subpatent infections from the Amazon region in Brazil

Asymptomatic Plasmodium infection is a new challenge for public health in the American region. The polymerase chain reaction (PCR) is the best method for diagnosing subpatent parasitemias. In endemic areas, blood collection is hampered by geographical distances and deficient transport and storage conditions of the samples. Because DNA extraction from blood collected on filter paper is an efficient method for molecular studies in high parasitemic individuals, we investigated whether the technique could be an alternative for Plasmodium diagnosis among asymptomatic and pauciparasitemic subjects. In this report we compared three different methods (Chelex®-saponin, methanol and TRIS-EDTA) of DNA extraction from blood collected on filter paper from asymptomatic Plasmodium-infected individuals. Polymerase chain reaction assays for detection of Plasmodium species showed the best results when the Chelex®-saponin method was used. Even though the sensitivity of detection was approximately 66% and 31% for P. falciparum and P. vivax, respectively, this method did not show the effectiveness in DNA extraction required for molecular diagnosis of Plasmodium. The development of better methods for extracting DNA from blood collected on filter paper is important for the diagnosis of subpatent malarial infections in remote areas and would contribute to establishing the epidemiology of this form of infection.

Reparação Híbrida do Arco Aórtico: Experiência Inicial do Hospital de Santa Marta

Objectives: To retrospectively review the hybrid treatment of the aortic arch with supra-aortic debranching and endo- vascular stent-graft repair in a single institution. Methods: From 2007 to 2010, all patients submitted to aortic debranching procedures were entered into a prospective database analysis. For the present study, only patients with sealing zones 0 and 1, according to the Ishimaru classification, were included. Procedure-related morbimortality was analysed for the open and endovascular procedures. Results: During the study period, we electively performed 6 total aortic debranching and 4 partial aortic debranching procedures in 10 patients. According to the etiology the indications were: 6 aortic arch aneurysms, 2 post-dissection aneu- rysms, 1 false aneurysm and 1 type I endoleak following TEVAR. The proximal sealing zone was Ishimaru zone 0 in six patients and zone 1 in four patients. The TEVAR procedure was delayed in all patients with a completion success of 80% (1 patient died from ruptured aortic aneurysm; 1 patient denied the second procedure and was lost to follow-up). The 30d mortality rate was 10% (patient mentioned above). The main morbidity was: 1 axillar venous thrombosis, 1 case of subclinical myocardial infarction, 1 case of terminal renal insufficiency and 1 case of prolonged ventilation. No permanent cerebral or peripheral neurologic deficit was noted. Conclusions: The hybrid repair of the aortic arch is a feasible and reproducible procedure, and our results are similar to the previously published series. Medium and long-term results are necessary to confirm whether the technique can be regarded as a safe alternative to open surgery in high-risk patients.

Exploraty Multivariate Statistical Methods Applied to Pharmaceutical Industry CRM Data

Dissertação apresentada como requisito parcial para obtenção do grau de Mestre em Estatística e Gestão de Informação

EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES

Currently there are several methods to extract bacterial DNA based on different principles. However, the amount and the quality of the DNA obtained by each one of those methods is highly variable and microorganism dependent, as illustrated by coagulase-negative staphylococci (CoNS) which have a thick cell wall that is difficult to lyse. This study was designed to compare the quality and the amount of CoNS DNA, extracted by four different techniques: two in-house protocols and two commercial kits. DNA amount and quality determination was performed through spectrophotometry. The extracted DNA was also analyzed using agarose gel electrophoresis and by PCR. 267 isolates of CoNS were used in this study. The column method and thermal lyses showed better results with regard to DNA quality (mean ratio of A260/280 = 1.95) and average concentration of DNA (), respectively. All four methods tested provided appropriate DNA for PCR amplification, but with different yields. DNA quality is important since it allows the application of a large number of molecular biology techniques, and also it's storage for a longer period of time. In this sense the extraction method based on an extraction column presented the best results for CoNS.

Estudo e otimização de juntas do tipo Tpeel soldadas, adesivas e híbridas

A necessidade de utilizar métodos de ligação que melhor satisfaçam as necessidades de projeto tem causado a crescente utilização das juntas adesivas, em detrimento dos métodos tradicionais tais como a soldadura, ligações aparafusadas e rebitadas. A sua utilização em diversas aplicações industriais justifica-se pela redução de peso, redução de concentrações de tensões, isolamento acústico e melhor resistência à corrosão. Contudo, também apresentam desvantagens, como a necessidade de preparação das juntas, a fraca resistência a esforços de arrancamento e a complexidade da previsão da sua resistência. As juntas híbridas são obtidas por combinação de uma técnica tradicional com uma ligação adesiva. As juntas híbridas adesivas-soldadas obtêm-se através da combinação da ligação adesiva com a ligação soldada, sendo a soldadura de resistência por pontos a técnica de soldadura mais usada no fabrico deste tipo de juntas. A sinergia entre ligação adesiva e soldadura por pontos oferece vantagens competitivas em relação às ligações adesivas, tais como superior resistência e rigidez, e maior resistência ao arrancamento e à fadiga. No presente trabalho é apresentado um estudo experimental e numérico de juntas T-peel soldadas, adesivas e híbridas (adesivas-soldadas) solicitadas ao arrancamento. Considerouse o adesivo frágil Araldite® AV138 e os adesivos dúcteis Araldite® 2015 e Sikaforce® 7752 e aderentes de aço (C45E). Foi realizada uma análise dos valores experimentais e efetuada uma comparação destes valores com os resultados obtidos pelo Método de Elementos Finitos (MEF) no software ABAQUS®, que incluiu uma análise de tensões na camada de adesivo e previsão do comportamento das juntas por MDC. Observou-se que, dos três adesivos em estudo, o adesivo Sikaforce® 7752 é o que apresenta o melhor desempenho na ligação de juntas T-peel. A boa concordância entre os resultados experimentais e numéricos permitiu validar a utilização de MDC para previsão da resistência de juntas T-peel adesivas e híbridas. Assim, o presente trabalho representa uma base para posterior aplicação no projeto deste tipo de ligação, com as vantagens decorrentes na redução do tempo de projeto e maior facilidade de otimização.

Parallelization of web processing services on cloud computing: A case study of Geostatistical Methods

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.

EVALUATION OF METHODS IN DETECTING VANCOMYCIN MIC AMONG MRSA ISOLATES AND THE CHANGES IN ACCURACY RELATED TO DIFFERENT MIC VALUES

INTRODUCTION: Methicillin-Resistant Staphylococcus aureus (MRSA) presenting reduced susceptibility to vancomycin has been associated to therapeutic failure. Some methods used by clinical laboratories may not be sufficiently accurate to detect this phenotype, compromising results and the outcome of the patient. OBJECTIVES: To evaluate the performance of methods in the detection of vancomycin MIC values among clinical isolates of MRSA. MATERIAL AND METHODS: The Vancomycin Minimal Inhibitory Concentration was determined for 75 MRSA isolates from inpatients of Mãe de Deus Hospital, Porto Alegre, Brazil. The broth microdilution (BM) was used as the gold-standard technique, as well as the following methods: E-test® strips (BioMérieux), M.I.C.E® strips (Oxoid), PROBAC® commercial panel and the automated system MicroScan® (Siemens). Besides, the agar screening test was carried out with 3 µg/mL of vancomycin. RESULTS: All isolates presented MIC ≤ 2 µg/mL for BM. E-test® had higher concordance (40%) in terms of global agreement with the gold standard, and there was not statistical difference among E-test® and broth microdilution results. PROBAC® panels presented MICs, in general, lower than the gold-standard panels (58.66% major errors), while M.I.C.E.® MICs were higher (67.99% minor errors). CONCLUSIONS: For the population of MRSA in question, E-test® presented the best performance, although with a heterogeneous accuracy, depending on MIC values.

Position Statement on Bioresorbable Vascular Scaffolds in Portugal

BACKGROUND: Bioresorbable vascular scaffolds (BVS) were recently approved for percutaneous coronary intervention in Europe. The aim of this position statement is to review the information and studies on available BVS, to stimulate discussion on their use and to propose guidelines for this treatment option in Portugal. METHODS AND RESULTS: A working group was set up to reach a consensus based on current evidence, discussion of clinical case models and individual experience. The evidence suggests that currently available BVS can produce physiological and clinical improvements in selected patients. There are encouraging data on their durability and long-term safety. Initial indications were grouped into three categories: (a) consensual and appropriate - young patients, diabetic patients, left anterior descending artery, long lesions, diffuse disease, and hybrid strategy; (b) less consensual but possible - small collateral branches, stabilized acute coronary syndromes; and (c) inappropriate - left main disease, tortuosity, severe calcification. CONCLUSION: BVS are a viable treatment option based on the encouraging evidence of their applicability and physiological and clinical results. They should be used in appropriate indications and will require technical adaptations. Outcome monitoring and evaluation is essential to avoid inappropriate use. It is recommended that medical societies produce clinical guidelines based on high-quality registries as soon as possible.

Strength and damage analysis of composite-aluminium adhesively-bonded single-lap joints

With the need to find an alternative way to mechanical and welding joints, and at the same time to overcome some limitations linked to these traditional techniques, adhesive bonds can be used. Adhesive bonding is a permanent joining process that uses an adhesive to bond the components of a structure. Composite materials reinforced with fibres are becoming increasingly popular in many applications as a result of a number of competitive advantages. In the manufacture of composite structures, although the fabrication techniques reduce to the minimum by means of advanced manufacturing techniques, the use of connections is still required due to the typical size limitations and design, technological and logistical aspects. Moreover, it is known that in many high performance structures, unions between composite materials with other light metals such as aluminium are required, for purposes of structural optimization. This work deals with the experimental and numerical study of single lap joints (SLJ), bonded with a brittle (Nagase Chemtex Denatite XNRH6823) and a ductile adhesive (Nagase Chemtex Denatite XNR6852). These are applied to hybrid joints between aluminium (AL6082-T651) and carbon fibre reinforced plastic (CFRP; Texipreg HS 160 RM) adherends in joints with different overlap lengths (LO) under a tensile loading. The Finite Element (FE) Method is used to perform detailed stress and damage analyses allowing to explain the joints’ behaviour and the use of cohesive zone models (CZM) enables predicting the joint strength and creating a simple and rapid design methodology. The use of numerical methods to simulate the behaviour of the joints can lead to savings of time and resources by optimizing the geometry and material parameters of the joints. The joints’ strength and failure modes were highly dependent on the adhesive, and this behaviour was successfully modelled numerically. Using a brittle adhesive resulted in a negligible maximum load (Pm) improvement with LO. The joints bonded with the ductile adhesive showed a nearly linear improvement of Pm with LO.

COMPARISON BETWEEN FOUR USUAL METHODS OF IDENTIFICATION OF Candida SPECIES

SUMMARY Infection by Candidaspp. is associated with high mortality rates, especially when treatment is not appropriate and/or not immediate. Therefore, it is necessary to correctly identify the genus and species of Candida. The aim of this study was to compare the identification of 89 samples of Candida spp. by the manual methods germ tube test, auxanogram and chromogenic medium in relation to the ID 32C automated method. The concordances between the methods in ascending order, measured by the Kappa index were: ID 32C with CHROMagar Candida(κ = 0.38), ID 32C with auxanogram (κ = 0.59) and ID 32C with germ tube (κ = 0.9). One of the species identified in this study was C. tropicalis,which demonstrated a sensitivity of 46.2%, a specificity of 95.2%, PPV of 80%, NPV of 81.1%, and an accuracy of 80.9% in tests performed with CHROMagar Candida;and a sensitivity of 76.9%, a specificity of 96.8%, PPV of 90.9%, NPV of 91%, and an accuracy of 91% in the auxanogram tests. Therefore, it is necessary to know the advantages and limitations of methods to choose the best combination between them for a fast and correct identification of Candidaspecies.

Comparison of Methods and Co-Registration Maps of EEG and fMRI in Occipital Lobe Epilepsy

Clinically childhood occipital lobe epilepsy (OLE) manifests itself with distinct syndromes. The traditional EEG recordings have not been able to overcome the difficulty in correlating the ictal clinical symptoms to the onset in particular areas of the occipital lobes. To understand these syndromes it is important to map with more precision the epileptogenic cortical regions in OLE. Experimentally, we studied three idiopathic childhood OLE patients with EEG source analysis and with the simultaneous acquisition of EEG and fMRI, to map the BOLD effect associated with EEG spikes. The spatial overlap between the EEG and BOLD results was not very good, but the fMRI suggested localizations more consistent with the ictal clinical manifestations of each type of epileptic syndrome. Since our first results show that by associating the BOLD effect with interictal spikes the epileptogenic areas are mapped to localizations different from those calculated from EEG sources and that by using different EEG/fMRI processing methods our results differ to some extent, it is very important to compare the different methods of processing the localization of activation and develop a good methodology for obtaining co-registration maps of high resolution EEG with BOLD localizations.

COMPARISON OF PERMANENT STAINING METHODS FOR THE LABORATORY DIAGNOSIS OF TRICHOMONIASIS

Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in the world. The diagnosis is based on wet mount preparation and direct microscopy on fixed and stained clinical specimens. The aim of this study was to compare the performance of different fixing and staining techniques used in the detection of T. vaginalis in urine. The smears were fixed and submitted to different methods of permanent staining and then, the morphological aspects of the parasites were analyzed and compared. The Papanicolaou staining with ethanol as the fixative solution showed to be the best method of permanent staining. Our data suggest that staining techniques in association with wet mount examination of fresh specimens contribute to increase the sensitivity in the diagnosis of trichomoniasis.

Numerical methods and tangible interfaces for pollutant dispersion simulation

Dissertação apresentada para obtenção do Grau de Doutor em Engenharia do Ambiente, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Comparison of vigilance methods in three houses with different Triatoma infestans densities

Através da prova de 7 dias foi estudado o grau de resistência do Plasmodium falciparum à cloroquina, amodiaquina e sulfadoxina-pirimetamina em Porto Velho, Estado de Rondônia, Brasil. Não se observaram diferenças significativas nas médias de parasitas nos dias de seguimento e nas proporções de resistência entre os três medicamentos testados, fazendo com que os autores recomendem a manutenção das 4-aminoquinoleínas como drogas a serem usadas atualmente em infecções não graves por P. falciparum na área de Porto Velho.

Bullwhip effect, in production control: a comparison between traditional methods and LPS

21th Annual Conference of the International Group for Lean Construction (IGLC 21), July 2013, Fortaleza, Brazil