COMPARISON BETWEEN AUTOMATED SYSTEM AND PCR-BASED METHOD FOR IDENTIFICATION AND ANTIMICROBIAL SUSCEPTIBILITY PROFILE OF CLINICAL Enterococcus spp

Enterococci are increasingly responsible for nosocomial infections worldwide. This study was undertaken to compare the identification and susceptibility profile using an automated MicrosScan system, PCR-based assay and disk diffusion assay of Enterococcus spp. We evaluated 30 clinical isolates of Enterococcus spp. Isolates were identified by MicrosScan system and PCR-based assay. The detection of antibiotic resistance genes (vancomycin, gentamicin, tetracycline and erythromycin) was also determined by PCR. Antimicrobial susceptibilities to vancomycin (30 µg), gentamicin (120 µg), tetracycline (30 µg) and erythromycin (15 µg) were tested by the automated system and disk diffusion method, and were interpreted according to the criteria recommended in CLSI guidelines. Concerning Enterococcus identification the general agreement between data obtained by the PCR method and by the automatic system was 90.0% (27/30). For all isolates of E. faecium and E. faecalis we observed 100% agreement. Resistance frequencies were higher in E. faecium than E. faecalis. The resistance rates obtained were higher for erythromycin (86.7%), vancomycin (80.0%), tetracycline (43.35) and gentamicin (33.3%). The correlation between disk diffusion and automation revealed an agreement for the majority of the antibiotics with category agreement rates of > 80%. The PCR-based assay, the van(A) gene was detected in 100% of vancomycin resistant enterococci. This assay is simple to conduct and reliable in the identification of clinically relevant enterococci. The data obtained reinforced the need for an improvement of the automated system to identify some enterococci.

HIGH SENSITIVE PCR METHOD FOR DETECTION OF PATHOGENIC Leptospira spp. IN PARAFFIN-EMBEDDED TISSUES

This study describes the development and application of a new PCR assay for the specific detection of pathogenic leptospires and its comparison with a previously reported PCR protocol. New primers were designed for PCR optimization and evaluation in artificially-infected paraffin-embedded tissues. PCR was then applied to post-mortem, paraffin-embedded samples, followed by amplicon sequencing. The PCR was more efficient than the reported protocol, allowing the amplification of expected DNA fragment from the artificially infected samples and from 44% of the post-mortem samples. The sequences of PCR amplicons from different patients showed >99% homology with pathogenic leptospires DNA sequences. The applicability of a highly sensitive and specific tool to screen histological specimens for the detection of pathogenic Leptospira spp. would facilitate a better assessment of the prevalence and epidemiology of leptospirosis, which constitutes a health problem in many countries.

Estudo numérico da adequação de diferentes tipos de leis coesivas na previsão da resistência de juntas adesivas por Modelos de Dano Coesivo

O uso de ligações adesivas aumentou significativamente nos últimos anos e é hoje em dia uma técnica de ligação dominante na indústria aeronáutica e automóvel. As ligações adesivas visam substituir os métodos tradicionais de fixação mecânicos na união de estruturas. A melhoria ao longo dos anos de vários modelos de previsão de dano, nomeadamente através do Método de Elementos Finitos (MEF), tem ajudado ao desenvolvimento desta técnica de ligação. Os Modelos de Dano coesivo (MDC), usados em conjunto com MEF, são uma ferramenta viável para a previsão de resistência de juntas adesivas. Os MDC combinam critérios da resistência dos materiais para a iniciação do dano e conceitos da mecânica da fratura para a propagação da fenda. Existem diversas formas de leis coesivas possíveis de aplicar em simulações por MDC, em função do comportamento expectável dos materiais que estão a ser simulados. Neste trabalho, estudou-se numericamente o efeito de diversas formas de leis coesivas na previsão no comportamento de juntas adesivas, nomeadamente nas curvas forçadeslocamento (P-) de ensaios Double-Cantilever Beam para caracterização à tração e ensaios End-Notched Flexure para caraterização ao corte. Também se estudou a influência dos parâmetros coesivos à tração e corte nas curvas P- dos referidos ensaios. Para o Araldite®AV138 à tração e ao corte, a lei triangular é a que melhor prevê o comportamento do adesivo. Para a previsão da resistência de ambos os adesivos Araldite® 2015 e SikaForce® 7752, a lei trapezoidal é a que melhor se adequa, confirmando assim que esta lei é a que melhor caracteriza o comportamento de dano de adesivos tipicamente dúcteis. O estudo dos parâmetros revelou influência distinta na previsão do comportamento das juntas, embora com bastantes semelhanças entre os diferentes tipos de adesivos.

Ponseti Method: Does Age at the Beginning of Treatment Make a Difference?

The Ponseti method is reportedly effective for treating clubfoot in children up to 9 years of age. However, whether age at the beginning of treatment influences the rate of successful correction and the rate of relapse is unknown. We therefore retrospectively reviewed 68 consecutive children with 102 idiopathic clubfeet treated by the Ponseti technique in four Portuguese hospitals. We followed patients a minimum of 30 months (mean, 41.4 months; range, 30–61 months). The patients were divided into two groups according to their age at the beginning of treatment; Group I was younger than 6 months and Group II was older than 6 months. All feet(100%) were initially corrected and no feet required extensive surgery regardless of age at the beginning of treatment. There were no differences between Groups I and II in the number of casts, tenotomies, success in terms of rate of initial correction, rate of recurrence, and rate of tibialis anterior transference. The rate of the Ponseti method in avoiding extensive surgery was 100% in Groups I and II; relapses occurred in 8% of the feet in younger and older children. Level of Evidence: Level II, prognostic study. See the Guidelines for Authors for a complete description of levels of evidence.

Aplicação do modelo escoras e tirantes a vigas parede de acordo com o Eurocódigo 2:2010

O Modelo Escoras e Tirantes surgiu no início do Século XX quando Ritter e Mörsch estabeleceram a analogia entre uma treliça clássica e uma viga de betão armado. Desde então pesquisadores têm estudado esse modelo como método de dimensionamento. A partir dos anos 90 várias normas apresentaram a utilização do modelo escoras e tirantes como relevante no dimensionamento de elementos de betão armado. Os critérios de segurança do Modelo Escoras e Tirantes são neste trabalho explicados de acordo com o Eurocódigo 2:2010, e comparadas com as normas NBR 6118:2014 e ACI 318:2011. É unanime dizer que a utilização do método torna-se mais vantajosa para regiões de descontinuidade. Em todos os elementos de betão armado, o Método Escoras e Tirantes são representações dos campos de tensão idealizados por elementos comprimidos e tracionados. Para a definição destes elementos é proposto o processo de “caminho de carga” em que conhecidas as tensões elásticas e suas direções principais utilizando o Método dos Elementos Finitos o modelo das escoras e tirantes é de fácil concepção. É também possível a definição deste a partir de modelos padrão já concebidos para determinados tipos de elementos estruturais de betão armado. Para o elemento descontinuo viga-parede estudado foram apresentados cinco modelos de cálculo até otimizar a solução validando as tensões com o Método dos Elementos Finitos. Em todos os modelos foram analisadas a definição do Modelo Escoras e Tirantes, a resistência das escoras, dos tirantes e dos nós até chegar à solução construtiva da viga-parede. Concluiu-se que a definição do modelo é a chave do dimensionamento.

Preparation of gas-filled porous microparticles (GPPs) and microbubbles (MBs) by PGSS method

Dissertation to obtain the Master Degree in Biotechnology

The sum-of-the-parts method: An international application

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Finance from the NOVA – School of Business and Economics

A Homemade Snare: An Alternative Method for Mechanical Removal of Dirofilaria Immitis in Dogs

Canine dirofilariosis is a life-threatening parasitic disease that is increasingly reported worldwide. Once diagnosed the main treatment goals are to improve the animal's clinical condition and to eliminate all life stages of the parasite with minimal posttreatment side effects. This can be achieved through mechanical, surgical, or chemotherapeutical approaches. Currently, manual extraction is the preferred method to remove adult heartworms due to its diminished invasiveness, reduced damage to the vascular endothelium, and shortened anaesthesia duration. However, it remains an expensive technique that can be highly traumatic. To address this issue, a nontraumatic homemade catheter-guided snare was developed for heartworm removal by adapting and folding a 0.014-inch coronary wire (BMW, Abbott Vascular). Transvenous heartworm extraction was performed on a dog severely infected with adult heartworms by inserting the modified snare into a 6-F Judkins right coronary guiding catheter BMW (Cordis) and advancing it into the right ventricle under fluoroscopic guidance. Fifteen adult specimens of Dirofilaria immitis were successfully extracted from the pulmonary artery and right ventricle without complications. To assure the death of both larvae and adults, postoperative treatment was successfully managed using ivermectin, doxycycline, and melarsomine, with no recurrence after surgery.

What is the best accounting method for financial assets?

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Finance from the NOVA – School of Business and Economics

Speed control of induction machine based on direct torque control method

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

Atomic force microscopy assisted with electron and ion beam microscopy for the direct measurement of biostructures and DNA samples

Dissertation to obtain a Master degree in Biotechnology

A nanopore-based stochastic detection method: Single molecule characterisation of nanoparticles using ()- hemolysin

Dissertation presented to obtain the Ph.D degree in Chemistry.

In-house ELISA method to analyze anti-Trypanosoma cruzi IgG reactivity for differential diagnosis and evaluation of Chagas disease morbidity

INTRODUCTION: The goal was to develop an in-house serological method with high specificity and sensitivity for diagnosis and monitoring of Chagas disease morbidity. METHODS: With this purpose, the reactivities of anti-T. cruzi IgG and subclasses were tested in successive serum dilutions of patients from Berilo municipality, Jequitinhonha Valley, Minas Gerais, Brazil. The performance of the in-house ELISA was also evaluated in samples from other relevant infectious diseases, including HIV, hepatitis C (HCV), syphilis (SYP), visceral leishmaniasis (VL), and American tegumentary leishmaniasis (ATL), and noninfected controls (NI). Further analysis was performed to evaluate the applicability of this in-house methodology for monitoring Chagas disease morbidity into three groups of patients: indeterminate (IND), cardiac (CARD), and digestive/mixed (DIG/Mix), based on their clinical status. RESULTS: The analysis of total IgG reactivity at serum dilution 1:40 was an excellent approach to Chagas disease diagnosis (100% sensitivity and specificity). The analysis of IgG subclasses showed cross-reactivity, mainly with NI, VL, and ATL, at all selected serum dilutions. Based on the data analysis, the IND group displayed higher IgG3 levels and the DIG/Mix group presented higher levels of total IgG as compared with the IND and CARD groups. CONCLUSIONS: These findings demonstrated that methodology presents promising applicability in the analysis of anti-T. cruzi IgG reactivity for the differential diagnosis and evaluation of Chagas disease morbidity.