Process engineering of liver cells for drug testing applications

Dissertação para obtenção do Grau de Doutor em Bioengenharia

HIV Voluntary Testing among Portuguese Women Attending Family Planning Clinics: Implications for HIV Prevention Education and Testing

Objectives: The aim of this article is to analyze the factors associated with HIV testing among 767 sexually active women. Methods: Participants were administered several self-report questionnaires that assessed behavioral and psychosocial measures. Results: Overall, 59.8% of the participants reported ever having tested for HIV. Results show that higher levels of education, being pregnant or having been pregnant, concern about AIDS, AIDS knowledge, self-efficacy in condom negotiation and perception of no risk in partner significantly predicted the likelihood of testing among women. Attending the mass was negatively associated with HIV testing. Conclusions: These findings provide information that can be used in the development of a focused gender sensitive HIV prevention program to increase HIV testing.

A filter inexact-restoration method for nonlinear programming

A new iterative algorithm based on the inexact-restoration (IR) approach combined with the filter strategy to solve nonlinear constrained optimization problems is presented. The high level algorithm is suggested by Gonzaga et al. (SIAM J. Optim. 14:646–669, 2003) but not yet implement—the internal algorithms are not proposed. The filter, a new concept introduced by Fletcher and Leyffer (Math. Program. Ser. A 91:239–269, 2002), replaces the merit function avoiding the penalty parameter estimation and the difficulties related to the nondifferentiability. In the IR approach two independent phases are performed in each iteration, the feasibility and the optimality phases. The line search filter is combined with the first one phase to generate a “more feasible” point, and then it is used in the optimality phase to reach an “optimal” point. Numerical experiences with a collection of AMPL problems and a performance comparison with IPOPT are provided.

Isolation and antifungal susceptibility testing of Trichosporon asahii in Ceará, Brazil

Trichosporon spp. are yeasts capable of causing invasive disease, which mainly affect immunocompromised patients. A clinical strain of T. asahii was isolated from the blood cultures of patients admitted to the General Hospital of Fortaleza. Susceptibility tests were conducted by disk diffusion and broth microdilution. The isolated strain of T. asahii was resistant to fluconazole. The patient used amphotericin B and caspofungin in order to facilitate the microbiological cure. It was the first isolation and identification of T. asahii in blood culture in Ceará, Brazil.

In vitro synergism of simvastatin and fluconazole against Candida species

Systemic fungal infections are responsible for high mortality rates. Several species of fungi may be involved, but Candida spp. is the most prevalent. Simvastatin is used to lower cholesterol and also exhibits antifungal action. The aim of this study was to evaluate the synergistic action of simvastatin with fluconazole against strains of Candida spp. Susceptibility testing was performed according to protocol M27-A3, by broth microdilution method and the synergistic effect of simvastatin and fluconazole was calculated based on FICI (Fractional Inhibitory Concentration Index). Eleven strains were evaluated, and simvastatin showed a synergistic effect with fluconazole against 10 (91%) of the Candida spp. strains tested. Simvastatin may be a valuable drug in the treatment of systemic infections caused by Candida spp.

Comparison of five methods for oxacillin susceptibility testing of Staphylococcus aureus isolates from cystic fibrosis patients

Methicillin-resistant Staphylococcus aureus (MRSA) are now a worldwide problem. Cystic fibrosis (CF) patients are commonly colonized and infected by MRSA. Accurate oxacillin susceptibility testing is mandatory for the adequate management of these patients. We performed a comparison of the accuracy of different tests in CF isolates, including methicillin-susceptible S. aureus and MRSA with different SCCmec types, and using the mecA gene as the gold-standard. The sensitivity and specificity of oxacillin disc, Etest, and oxacillin agar screening plate were 100%. Sensitivity of the cefoxitin disc was 85% and specificity was 100%. For clinically relevant isolates, laboratories may consider the use of a combination of two phenotypic methods.

Aplicação de um método de correlação de imagem para a determinação da tenacidade à fratura em corte de adesivos estruturais

Qualquer estrutura hoje em dia deve ser resistente, robusta e leve, o que aumentou o interesse industrial e investigação nas ligações adesivas, nomeadamente pela melhoria das propriedades de resistência e fratura dos materiais. Com esta técnica de união, o projeto de estruturas pode ser orientado para estruturas mais leves, não só em relação à economia direta de peso relativamente às juntas aparafusas ou soldadas, mas também por causa da flexibilidade para ligar materiais diferentes. Em qualquer área da indústria, a aplicação em larga escala de uma determinada técnica de ligação supõe que estão disponíveis ferramentas confiáveis para o projeto e previsão da rotura. Neste âmbito, Modelos de Dano Coesivo (MDC) são uma ferramenta essencial, embora seja necessário estimar as leis MDC do adesivo à tração e corte para entrada nos modelos numéricos. Este trabalho avalia o valor da tenacidade ao corte (GIIC) de juntas coladas para três adesivos com ductilidade distinta. O trabalho experimental consiste na caracterização à fratura ao corte da ligação adesiva por métodos convencionais e pelo Integral-J. Além disso, pelo integral-J, é possível definir a forma exata da lei coesiva. Para o integral-J, é utilizado um método de correlação de imagem digital anteriormente desenvolvido para a avaliação do deslocamento ao corte do adesivo na extremidade da fenda (δs) durante o ensaio, acoplado a uma sub-rotina em Matlab® para a extração automática de δs. É também apresentado um trabalho numérico para avaliar a adequabilidade de leis coesivas triangulares aproximadas em reproduzir as curvas força-deslocamento (P-δ) experimentais dos ensaios ENF. Também se apresenta uma análise de sensibilidade para compreender a influência dos parâmetros coesivos nas previsões numéricas. Como resultado deste trabalho, foram estimadas experimentalmente as leis coesivas de cada adesivo pelo método direto, e numericamente validadas, para posterior previsão de resistência em juntas adesivas. Em conjunto com a caraterização à tração destes adesivos, é possível a previsão da rotura em modo-misto.

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.

HEPATITIS B VACCINATION COVERAGE AND POSTVACCINATION SEROLOGIC TESTING AMONG MEDICAL STUDENTS AT A PUBLIC UNIVERSITY IN BRAZIL

The aim of this cross-sectional study was to determine the hepatitis B vaccination coverage among medical students at a public university in Rio de Janeiro, Brazil, and their compliance with the postvaccination serologic testing recommendations. Of the total of 858 students, 675 (78.7%) participated in the study. Among the participants, 48.9% (95% CI: 45.1% to 52.7%) were vaccinated against hepatitis B (received ≥ 3 doses of the vaccine), 31.6% were not (received 0, 1 or 2 doses), and 19.6% did not know their vaccination status. Hepatitis B vaccination coverage increased from 26.0% among first-year students to 70.6% among sixth-year students while the prevalence of unknown vaccination status decreased from 39.7% among first-year students to 2.4% among sixth-year students. The frequency of unvaccinated students ranged from 23.7% among fifth-year students to 34.4% among first-year students. Only 34.8% of the vaccinated students performed the anti-HBs testing after vaccination. Among these medical students, we found a low adherence to the hepatitis B vaccination and to the postvaccination serologic testing. A comprehensive hepatitis B immunization program should be offered to students at this medical school.

In vitro SCREENING ANTIBACTERIAL ACTIVITY OF Bidens pilosa LINNÉ AND Annona crassiflora MART. AGAINST OXACILLIN RESISTANT Staphylococcus aureus (ORSA) FROM THE AERIAL ENVIRONMENT AT THE DENTAL CLINIC

Currently multiresistant Staphylococcus aureus is one common cause of infections with high rates of morbidity and mortality worldwide, which directs scientific endeavors in search for novel antimicrobials. In this study, nine extracts from Bidens pilosa (root, stem, flower and leaves) and Annona crassiflora (rind fruit, stem, leaves, seed and pulp) were obtained with ethanol: water (7:3, v/v) and their in vitro antibacterial activity evaluated through both the agar diffusion and broth microdilution methods against 60 Oxacillin Resistant S. aureus (ORSA) strains and against S. aureus ATCC6538. The extracts from B. pilosa and A. crassiflora inhibited the growth of the ORSA isolates in both methods. Leaves of B. pilosa presented mean of the inhibition zone diameters significantly higher than chlorexidine 0.12% against ORSA, and the extracts were more active against S. aureus ATCC (p < 0.05). Parallel, toxicity testing by using MTT method and phytochemical screening were assessed, and three extracts (B. pilosa, root and leaf, and A. crassiflora, seed) did not evidence toxicity. On the other hand, the cytotoxic concentrations (CC50 and CC90) for other extracts ranged from 2.06 to 10.77 mg/mL. The presence of variable alkaloids, flavonoids, tannins and saponins was observed, even though there was a total absence of anthraquinones. Thus, the extracts from the leaves of B. pilosa revealed good anti-ORSA activity and did not exhibit toxicity.

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.

Avaliação numérica e experimental da carga admissível de vigas compostas para monocarris de sistemas de elevação

Este trabalho foi realizado com o apoio, e em colaboração da empresa Flexcrane, que constrói e comercializa soluções flexíveis de pontes rolantes para cargas ligeiras. Com cargas máximas de 2 toneladas estas pontes e seus caminhos de rolamento são construídas essencialmente enformados a frio. O objetivo deste trabalho é a verificação numérica e experimental das cargas que este tipo de estrutura suporta, nas suas diferentes configurações. Numa aproximação por engenharia inversa, com a realização deste trabalho, foi possível determinar a carga admissível das vigas compostas utilizadas, assim como o seu comportamento quando em carga. Para obtenção destes resultados foram realizadas simulações computacionais recorrendo ao método de elementos finitos, utilizando para isso o software Ansys Workbench e ainda ensaios experimentais que permitiram validar os resultados obtidos numericamente e consolidar o conhecimento sobre o comportamento desta solução comercial. Por ser de construção modular, são possíveis diferentes combinações para o produto final, por esta razão foram estudados numericamente diferentes configurações possíveis, com e sem reforços. Foi realizada a validação experimental para a configuração mais usual, fazendo recurso a um dispositivo para ensaio de estruturas e componentes, pertencente ao Isep. Foi ainda utilizada a técnica extensométrica para avaliar as tensões em pontos selecionados e compara-los com os obtidos numericamente. Concluiu-se pelo ótimo desempenho dos produtos Flexcrane.

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.

Development Process and Cognitive Testing of CARATkids - Control of Allergic Rhinitis and Asthma Test for Children

Background: Allergic rhinitis and asthma (ARA) are chronic inflammatory diseases of the airways that often coexist in children. The only tool to assess the ARA control, the Control of Allergic Rhinitis and Asthma Test (CARAT) is to be used by adults. We aimed to develop the Pediatric version of Control of Allergic Rhinitis and Asthma Test (CARATkids) and to test its comprehensibility in children with 4 to 12 years of age. Methods: The questionnaire development included a literature review of pediatric questionnaires on asthma and/or rhinitis control and two consensus meetings of a multidisciplinary group. Cognitive testing was carried out in a cross-sectional qualitative study using cognitive interviews. Results: Four questionnaires to assess asthma and none to assess rhinitis control in children were identified. The multidisciplinary group produced a questionnaire version for children with 17 questions with illustrations and dichotomous (yes/no) response format. The version for caregivers had 4-points and dichotomous scales. Twenty-nine children, 4 to 12 years old, and their caregivers were interviewed. Only children over 6 years old could adequately answer the questionnaire. A few words/expressions were not fully understood by children of 6 to 8 years old. The drawings illustrating the questions were considered helpful by children and caregivers. Caregivers considered the questionnaire complete and clear and preferred dichotomous over the 4-points scales. The proportion of agreement between children and their caregivers was 61%. The words/expressions that were difficult to understand were amended. Conclusion: CARATkids, the first questionnaire to assess a child’s asthma and rhinitis control was developed and its content validity was assured. Cognitive testing showed that CARATKids is well-understood by children 6 to 12 years old. The questionnaire’s measurement properties can now be assessed in a validation study.