Comparison of ion pair and buffered mobile phase chromatography of haloacetic acids using direct and indirect UV and fluorescence detection

Reversed-phase high performance liquid chromatographic methods for the analysis of Haloacetic acids have been developed and compared to conventional direct detection methods. Haloacetic acids commonly found in drinking water, including monochloro-, dichloro-, bromo-, iodo- and trichloroacetic acids- have been studied. The ion pairing agent benzyltributylammonium ion was studied in detail using indirect UV and indirect fluorescence detection. Five different competing ions were evaluated to decrease analysis times and lower the detection limit by this new method. The direct detection method utilized an ammonium sulfate buffer and UV detection yielding a detection limit of 100 ppb. The indirect method developed has the advantage of being able to simultaneously analyze UV and non-UV absorbing ions and molecules but requires long equilibration times and demonstrated lower sensitivity than the direct method. ^

The development of direct ultra-fast PCR for forensic genotyping using short channel microfluidic systems with enhanced sieving matrices

There are situations in which it is very important to quickly and positively identify an individual. Examples include suspects detained in the neighborhood of a bombing or terrorist incident, individuals detained attempting to enter or leave the country, and victims of mass disasters. Systems utilized for these purposes must be fast, portable, and easy to maintain. The goal of this project was to develop an ultra fast, direct PCR method for forensic genotyping of oral swabs. The procedure developed eliminates the need for cellular digestion and extraction of the sample by performing those steps in the PCR tube itself. Then, special high-speed polymerases are added which are capable of amplifying a newly developed 7 loci multiplex in under 16 minutes. Following the amplification, a postage stamp sized microfluidic device equipped with specially designed entangled polymer separation matrix, yields a complete genotype in 80 seconds. The entire process is rapid and reliable, reducing the time from sample to genotype from 1-2 days to under 20 minutes. Operation requires minimal equipment and can be easily performed with a small high-speed thermal-cycler, reagents, and a microfluidic device with a laptop. The system was optimized and validated using a number of test parameters and a small test population. The overall precision was better than 0.17 bp and provided a power of discrimination greater than 1 in 106. The small footprint, and ease of use will permit this system to be an effective tool to quickly screen and identify individuals detained at ports of entry, police stations and remote locations. The system is robust, portable and demonstrates to the forensic community a simple solution to the problem of rapid determination of genetic identity.

Material Evaluation of an Elastomer, Epoxy and Lightweight Concrete Rail Attachment System for Direct Fixation Light Rail Applications

Thesis (Master's)--University of Washington, 2016-08

Chemical composition, acetylcholinesterase inhibitory and antifungal activities of Pera glabrata (Schott) Baill. (Euphorbiaceae)

Pera glabrata (Schott) Baill. was selected for this study after showing a preliminary positive result in a screening of Atlantic Forest plant species in the search for acetylcholinesterase inhibitors and antifungal compounds. The bioassays were conducted with crude ethanol extract of the leaves using direct bioautography method for acetylcholinesterase and antifungal activities. This extract was partitioned with hexane, chloroform and ethyl acetate solvents. The active chloroform fraction was submitted to silica gel chromatography column affording 12 groups. Caffeine, an alkaloid, which showed detection limits of 0.1 and 1.0 µg for anticholinesterasic and antifungal activities, respectively, was isolated from group nine. After microplate analyses, only groups four, nine, 10, 11 and 12 showed acetylcholinesterase inhibitory activity of 40% or higher. The group 12 was purified by preparative layer chromatography affording four sub-fractions. Two sub-fractions from this group were analyzed by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector. The first sub-fraction showed anticholinesterasic activity and contained two major compounds: 9-hydroxy-4-megastigmen-3-one (84%) and caffeine (6%). The second sub-fraction presented five major compounds identified as 9-hydroxy-4-megastigmen-3-one, isololiolide, (-) loliolide, palmitic acid and lupeol and did not show activity.

Consumo alimentar de micronutrientes entre pré-escolares no domicílio e em escolas de educação infantil do município de Caxias do Sul (RS)

OBJETIVO: Avaliar a ingestão alimentar de micronutrientes em pré-escolares no domicílio e em escolas de educação infantil públicas e particulares. MÉTODOS: Estudo transversal realizado com 362 pré-escolares entre dois e seis anos de idade, em Caxias do Sul (RS) Brasil, em 2007. A ingestão alimentar na escola foi avaliada por meio do método de pesagem direta individual, e no domicílio, por meio de registro alimentar realizado pelos pais ou responsáveis. Foi calculada a ingestão alimentar de cálcio, ferro, folato, vitamina A, vitamina C e zinco de acordo com o local da refeição e tipo de escola. RESULTADOS: Houve maior ingestão de alimentos contendo ferro, folato e vitamina C durante o período em que as crianças permaneceram na escola infantil, e maior ingestão de cálcio, vitamina A e zinco no domicílio. Houve significativamente maior ingestão de alimentos contendo ferro (p=0,03), folato (p=0,03), vitamina A (p<0,01) e vitamina C (p<0,01) pelas crianças da escola particular e maior ingestão de cálcio (p<0,01) e zinco (p<0,01) na escola pública. Quanto à prevalência de inadequação dos micronutrientes, as crianças não apresentaram risco deficiente para ingestão de ferro, folato, vitamina A e C e zinco, porém apenas 67,4% apresentaram ingestão de cálcio igual ou acima ao valor de referência. CONCLUSÃO: Os achados sugerem que o consumo de cálcio, vitamina A e zinco foi maior nos domicílios, apesar de as crianças permanecerem a maior parte do dia nas escolas. O consumo diário de micronutrientes de crianças de escolas públicas e particulares não diferiu significativamente, mesmo com diferenças nos cardápios.

Effect of processing induced particle alignment on the fracture toughness and fracture behavior of multiphase dental ceramics

Objective. To investigate the processing induced particle alignment on fracture behavior of four multiphase dental ceramics (one porcelain, two glass-ceramics and a glass-infiltrated-alumina composite). Methods. Disks (empty set12mm x 1.1 mm-thick) and bars (3 mm x 4 mm x 20 mm) of each material were processed according to manufacturer instructions, machined and polished. Fracture toughness (K(IC)) was determined by the indentation strength method using 3-point bending and biaxial flexure fixtures for the fracture of bars and disks, respectively. Microstructural and fractographic analyses were performed with scanning electron microscopy, energy dispersive spectroscopy and X-ray diffraction. Results. The isotropic microstructure of the porcelain and the leucite-based glass-ceramic resulted in similar fracture toughness values regardless of the specimen geometry. On the other hand, materials containing second-phase particles with high aspect ratio (lithium disilicate glass-ceramic and glass-infiltrated-alumina composite) showed lower fracture toughness for disk specimens compared to bars. For the lithium disilicate glass-ceramic disks, it was demonstrated that the occurrence of particle alignment during the heat-pressing procedure resulted in an unfavorable pattern that created weak microstructural paths during the biaxial test. For the glass-infiltrated-alumina composite, the microstructural analysis showed that the large alumina platelets tended to align their large surfaces perpendicularly to the direction of particle deposition during slip casting of green preforms. Significance. The fracture toughness of dental ceramics with anisotropic microstructure should be determined by means of biaxial testing, since it results in lower values. (C) 2009 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Desenvolvimento e padronização de um método para determinação da sensibilidade do mycobacterium tuberculosis a fármacos contra tuberculose

A resistência a fármacos antituberculose tem constituído uma grande ameaça ao controle da tuberculose em âmbito mundial. A sua detecção precoce permite ao médico instituir um esquema de tratamento mais adequado ao paciente e consequentemente quebrar a cadeia de transmissão dos bacilos. Os testes de sensibilidade a antimicrobianos atuais, embora eficientes, são caros e/ou demorados e/ou trabalhosos. Com base nesta premissa, nos propusemos a desenvolver e padronizar um método fenotípico direto para determinação da sensibilidade do Mycobacterium tuberculosis a antimicrobianos de primeira linha do tratamento da tuberculose. Para o desenvolvimento deste novo teste, utilizaram-se os princípios do método das proporções e do exame de cultura pelo método de Ogawa–Kudoh. O estudo foi dividido em duas fases. A primeira, caracterizada pelo desenvolvimento e padronização do método proposto e a segunda, pela análise da concordância entre o método desenvolvido e o método do MGIT (padrão-ouro). Na primeira fase, foram realizados diversos ensaios para definir: os volumes de absorção e de liberação de líquidos de diferentes tipos de swab, o meio de cultura, as concentrações dos antimicrobianos e o tempo de leitura/interpretação das culturas. Além disso, foi verificado se a amostra deveria ou não ser diluída. Com base nos resultados destes ensaios, padronizou-se o método com: swab comercial, em meio de cultura Ogawa- Kudoh contendo separadamente 0,2 μg/mL de isoniazida, 40,0 μg/mL de rifampicina, 10,0 μg/mL de estreptomicina e 500,0 μg/mL de ácido para-nitrobenzóico. Padronizou-se ainda a inoculação da amostra de escarro de forma direta, ou seja, sem diluir e a leitura/interpretação do resultado do teste no período entre 21 e 28 dias. A análise comparativa entre este método e o teste de sensibilidade a antimicrobianos no sistema MGIT realizada na segunda fase do projeto indicou um índice kappa igual a 1,000, ou seja, uma concordância muito boa em relação ao padrão-ouro. Diante desses resultados promissores, acreditamos que o método desenvolvido apresente um grande potencial para ser utilizado em laboratórios com pouca infra-estrutura, por ser de baixo custo, fácil execução e relativamente rápido.

Perceção visual na avaliação diagnóstica em mamografia: uma revisão sistemática

Introdução – Na avaliação diagnóstica em mamografia, o desempenho do radiologista pode estar sujeito a erros de diagnóstico. Objetivo – Descrever a importância da perceção visual na análise da mamografia, identificando os principais fatores que contribuem para a perceção visual do radiologista e que condicionam a acuidade diagnóstica. Metodologia – Estudo descritivo baseado numa revisão sistemática de literatura através da PubMed e da Science Direct. Foram incluídos 42 artigos que respeitavam, pelo menos, um dos critérios de inclusão no estudo. Para a seleção das referências foi utilizada a metodologia PRISMA, constituída por 4 fases: identificação, seleção preliminar, elegibilidade e estudos incluídos. Resultados – Na avaliação diagnóstica em mamografia, a perceção visual está intimamente relacionada com: 1) diferentes parâmetros visuais e da motilidade ocular (acuidade visual, sensibilidade ao contraste e à luminância e movimentos oculares); 2) com condições de visualização de uma imagem (iluminância da sala e luminância do monitor); e 3) fadiga ocular provocada pela observação diária consecutiva de imagens. Conclusões – A perceção visual pode ser influenciada por 3 categorias de erros observados: erros de pesquisa (lesões não são fixadas pela fóvea), erros de reconhecimento (lesões fixadas, mas não durante o tempo suficiente) e erros de decisão (lesões fixadas, mas não identificadas como suspeitas). Os estudos analisados sobre perceção visual, atenção visual e estratégia visual, bem como os estudos sobre condições de visualização não caracterizam a função visual dos observadores. Para uma avaliação correta da perceção visual em mamografia deverão ser efetuados estudos que correlacionem a função visual com a qualidade diagnóstica. ABSTRACT - Introduction – Diagnostic evaluation in mammography could be influenced by the radiologist performance that could be under diagnostic errors. Aims – To describe the importance of radiologist visual perception in mammographic diagnostic evaluation and to identify the main factors that contribute to diagnostic accuracy. Methods – In this systematic review 42 references were included based on inclusion criteria (PubMed and Science Direct). PRISMA method was used to select the references following 4 steps: identification, screening, eligibility and included references. Results – Visual perception in mammography diagnostic evaluation is related with: 1) visual parameters and ocular motility (visual acuity, contrast sensitivity and luminance and ocular movements); 2) image visualization environment (room iluminance and monitor luminance); and 3) eyestrain caused by image daily consecutive observation. Conclusions – Visual perception can be influenced by three errors categories: search errors (lesions are never looked at with high-resolution foveal vision), recognition errors (lesions are looked at, but not long enough to detect or recognize) and decision errors (lesions are looked at for long periods of time but are still missed). The reviewed studies concerning visual perception, visual attention, visual strategies and image visualization environment do not describe observer’s visual function. An accurate evaluation of visual perception in mammography must include visual function analysis.

Derivative-free optimization and filter methods to solve nonlinear constrained problems

In real optimization problems, usually the analytical expression of the objective function is not known, nor its derivatives, or they are complex. In these cases it becomes essential to use optimization methods where the calculation of the derivatives, or the verification of their existence, is not necessary: the Direct Search Methods or Derivative-free Methods are one solution. When the problem has constraints, penalty functions are often used. Unfortunately the choice of the penalty parameters is, frequently, very difficult, because most strategies for choosing it are heuristics strategies. As an alternative to penalty function appeared the filter methods. A filter algorithm introduces a function that aggregates the constrained violations and constructs a biobjective problem. In this problem the step is accepted if it either reduces the objective function or the constrained violation. This implies that the filter methods are less parameter dependent than a penalty function. In this work, we present a new direct search method, based on simplex methods, for general constrained optimization that combines the features of the simplex method and filter methods. This method does not compute or approximate any derivatives, penalty constants or Lagrange multipliers. The basic idea of simplex filter algorithm is to construct an initial simplex and use the simplex to drive the search. We illustrate the behavior of our algorithm through some examples. The proposed methods were implemented in Java.

Derivative-free nonlinear optimization filter simplex

The filter method is a technique for solving nonlinear programming problems. The filter algorithm has two phases in each iteration. The first one reduces a measure of infeasibility, while in the second the objective function value is reduced. In real optimization problems, usually the objective function is not differentiable or its derivatives are unknown. In these cases it becomes essential to use optimization methods where the calculation of the derivatives or the verification of their existence is not necessary: direct search methods or derivative-free methods are examples of such techniques. In this work we present a new direct search method, based on simplex methods, for general constrained optimization that combines the features of simplex and filter methods. This method neither computes nor approximates derivatives, penalty constants or Lagrange multipliers.

Optimizing potentiometric ionophore and electrode design for environmental on-site control of antibiotic drugs: Application to sulfamethoxazole

Potentiometric sensors are typically unable to carry out on-site monitoring of environmental drug contaminants because of their high limits of detection (LODs). Designing a novel ligand material for the target analyte and managing the composition of the internal reference solution have been the strategies employed here to produce for the first time a potentiometric-based direct reading method for an environmental drug contaminant. This concept has been applied to sulfamethoxazole (SMX), one of the many antibiotics used in aquaculture practices that may occur in environmental waters. The novel ligand has been produced by imprinting SMX on the surface of graphitic carbon nanostructures (CN) < 500 nm. The imprinted carbon nanostructures (ICN) were dispersed in plasticizer and entrapped in a PVC matrix that included (or not) a small amount of a lipophilic additive. The membrane composition was optimized on solid-contact electrodes, allowing near-Nernstian responses down to 5.2 μg/mL and detecting 1.6 μg/mL. The membranes offered good selectivity against most of the ionic compounds in environmental water. The best membrane cocktail was applied on the smaller end of a 1000 μL micropipette tip made of polypropylene. The tip was then filled with inner reference solution containing SMX and chlorate (as interfering compound). The corresponding concentrations were studied for 1 × 10−5 to 1 × 10−10 and 1 × 10−3 to 1 × 10−8 mol/L. The best condition allowed the detection of 5.92 ng/L (or 2.3 × 10−8 mol/L) SMX for a sub-Nernstian slope of −40.3 mV/decade from 5.0 × 10−8 to 2.4 × 10−5 mol/L.

Ensino direto da matemática funcional: estudo de caso

Neste estudo, focado na aprendizagem do manuseio do dinheiro, pretendeu-se que os alunos adquirissem competências que os habilitasse a um maior grau de independência e participação na vida em sociedade, desempenhando tarefas de cariz financeiro de forma mais independente, por exemplo, compra de produtos, pagamento de serviços e gestão do dinheiro. Para alcançar o pretendido, utilizou-se a metodologia do ensino direto, com tarefas estruturadas. Numa fase inicial o investigador prestava apoio constante aos alunos, que foi diminuindo gradualmente à medida que atingiam as competências relacionadas com o dinheiro. Na fase final, os alunos realizaram as tarefas propostas de forma autónoma. Construído como um estudo de caso, os dados foram recolhidos através de observação direta e de provas de monitorização. Os alunos começaram por realizar uma avaliação inicial para delinear a linha de base da intervenção. Posteriormente, foi realizada a intervenção baseada no ensino direto, com recurso ao computador, à calculadora, a provas de monitorização e ao manuseio de dinheiro. O computador foi utilizado na intervenção como tecnologia de apoio à aprendizagem, permitindo a realização de jogos interativos e consulta de materiais. No final da intervenção os alunos revelaram autonomia na resolução das tarefas, pois já tinham automatizado os processos matemáticas para saber manusear corretamente a moeda euro. O ensino direto auxiliou os alunos a reterem as competências matemáticas essenciais de manuseamento do dinheiro, compondo quantias, efetuando pagamentos e conferindo trocos, que muito podem contribuir para terem uma participação independente na vida em sociedade

Inflation forecasts using the tips yield curve

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

Numerical methods in multibody system dynamics

"Series: Solid mechanics and its applications, vol. 226"

Discordances between phylogenetic and morphological patterns in alpine leaf beetles attest to an intricate biogeographic history of lineages in postglacial Europe.

Pleistocene glacial and interglacial periods have moulded the evolutionary history of European cold-adapted organisms. The role of the different mountain massifs has, however, not been accurately investigated in the case of high-altitude insect species. Here, we focus on three closely related species of non-flying leaf beetles of the genus Oreina (Coleoptera, Chrysomelidae), which are often found in sympatry within the mountain ranges of Europe. After showing that the species concept as currently applied does not match barcoding results, we show, based on more than 700 sequences from one nuclear and three mitochondrial genes, the role of biogeography in shaping the phylogenetic hypothesis. Dating the phylogeny using an insect molecular clock, we show that the earliest lineages diverged more than 1 Mya and that the main shift in diversification rate occurred between 0.36 and 0.18 Mya. By using a probabilistic approach on the parsimony-based dispersal/vicariance framework (MP-DIVA) as well as a direct likelihood method of state change optimization, we show that the Alps acted as a cross-roads with multiple events of dispersal to and reinvasion from neighbouring mountains. However, the relative importance of vicariance vs. dispersal events on the process of rapid diversification remains difficult to evaluate because of a bias towards overestimation of vicariance in the DIVA algorithm. Parallels are drawn with recent studies of cold-adapted species, although our study reveals novel patterns in diversity and genetic links between European mountains, and highlights the importance of neglected regions, such as the Jura and the Balkanic range.