Periplasmic nitrate reductases: structural and spectroscopic studies

Dissertação apresentada para obtenção do grau de Doutor em Bioquímica, especialidade Bioquímica-Física, pela Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa

Contextual and individual indicators associated with the presence of teeth in adults

OBJECTIVE The objective of this study was to analyze whether socioeconomic conditions and the period of availability of fluoridated water are associated with the number of teeth present.METHODSThis cross-sectional study analyzed data from 1,720 adults between 20 and 59 years of age who resided in Florianópolis, SC, Southern Brazil, in 2009. The outcome investigated was the self-reported number of teeth present. The individual independent variables included gender, age range, skin color, number of years of schooling, and per capita household income. The duration of residence was used as a control variable. The contextual exposures included the period of availability of fluoridated water to the households and the socioeconomic variable for the census tracts, which was created from factor analysis of the tract’s mean income, education level, and percentage of households with treated water. Multilevel logistic regression was performed and inter-level interactions were tested.RESULTS Residents in intermediate and poorer areas and those with fluoridated water available for less time exhibited the presence of fewer teeth compared with those in better socioeconomic conditions and who had fluoridated water available for a longer period (OR = 1.02; 95%CI 1.01;1.02). There was an association between the period of availability of fluoridated water, per capita household income and number of years of education. The proportion of individuals in the poorer and less-educated stratum, which had fewer teeth present, was higher in regions where fluoridated water had been available for less time.CONCLUSIONS Poor socioeconomic conditions and a shorter period of availability of fluoridated water were associated with the probability of having fewer teeth in adulthood. Public policies aimed at reducing socioeconomic inequalities and increasing access to health services such as fluoridation of the water supply may help to reduce tooth loss in the future.

Chemical composition and antioxidante activity of Tunisian Origanum glandulosum Desf. essential oil and volatile oil obtain by supercritical CO2 extraction

Dried flowers and leaves of Origanum glandulosum Desf. were submitted to hydrodistillation (HD) and supercritical fluid extraction with CO2 (SFE). The essential oils isolated by HD and volatile oils obtained by SFE were analysed by GC and GC/MS. Total phenolics content and antioxidant effectiveness were performed. The main components of the essential oils from Bargou and Nefza were: p-cymene (40.4% and 39%), thymol (38.7% and 34.4%) and γ- terpinene (12.3% and 19.2%), respectively. The major components obtain by SFE in the volatile oil, from Bargou and Nefza, were: p-cymene (32.3% and 36.2%), thymol (41% and 40%) and γ-terpinene (20.3% and 13.3%). Total phenolic content, expressed in gallic acid equivalent (GAE) g kg-1 dry weight, varied from 12 to 27 g kg-1 dw, and the ability to scavenge the DPPH radicals, expressed by IC50 ranged from 44 to143 mg L-1.

UV-Vis spectroscopic study preferential solvation and intermolecular interactions in methanol/1-propanol/acetonitrile by means of solvatochromic probes

Solvatochromic UV-Vis shifts of four indicators (4-nitroaniline, 4-nitroanisole, 4-nitrophenol and N,N-dimethy-1-4-nitro aniline) have been measured at 298.15 K in the ternary mixture methano1/1-propanol/acetonitrile (MeOH/1-PrOH/MeCN) in a total of 22 mole fractions, along with 18 additional mole fractions for each of the corresponding binary mixtures, MeOH/1-PrOH, 1-PrOH/MeCN and MeOH/MeCN. These values, combined with our previous experimental results for 2,6-dipheny1-4-(2,4,6-triphenylpyridinium-1-yl)phenolate (Reichardt's betaine dye) in the same mixtures, permitted the computation of the Kamlet-Taft solvent parameters, alpha, beta, and pi*. The rationalization of the spectroscopic behavior of each probe within each mixture's whole mole fraction range was achieved through the use of the Bosch and Roses preferential solvation model. The applied model allowed the identification of synergistic behaviors in MeCN/alcohol mixtures and thus to infer the existence of solvent complexes in solution. Also, the addition of small amounts of MeCN to the binary mixtures was seen to cause a significant variation in pi*, whereas the addition of alcohol to MeCN mixtures always lead to a sudden change in a and The behavior of these parameters in the ternary mixture was shown to be mainly determined by the contributions of the underlying binary mixtures. (C) 2014 Elsevier B.V. All rights reserved.

Estudo comparativo de três câmaras de ionização para medição directa do equivalente de dose individual, a 10 mm de profundidade, Hp(10)

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Engenharia Biomédica

Extraction of volatile oil from aromatic plants with supercritical carbon dioxide: experiments and modeling

An overview of the studies carried out in our laboratories on supercritical fluid extraction (SFE) of volatile oils from seven aromatic plants: pennyroyal (Mentha pulegium L.), fennel seeds (Foeniculum vulgare Mill.), coriander (Coriandrum sativum L.), savory (Satureja fruticosa Beguinot), winter savory (Satureja montana L.), cotton lavender (Santolina chamaecyparisus) and thyme (Thymus vulgaris), is presented. A flow apparatus with a 1 L extractor and two 0.27 L separators was built to perform studies at temperatures ranging from 298 to 353 K and pressures up to 30.0 MPa. The best compromise between yield and composition compared with hydrodistillation (HD) was achieved selecting the optimum experimental conditions of extraction and fractionation. The major differences between HD and SFE oils is the presence of a small percentage of cuticular waxes and the relative amount of thymoquinone, an oxygenated monoterpene with important biological properties, which is present in the oils from thyme and winter savory. On the other hand, the modeling of our data on supercritical extraction of volatile oil from pennyroyal is discussed using Sovova's models. These models have been applied successfully to the other volatile oil extractions. Furthermore, other experimental studies involving supercritical CO2 carried out in our laboratories are also mentioned.

The effect of Schistosoma mansoni infection on child morbidity in the State in Bahia, Brazil: II — Analysis at the individual level

The present investigation was carried out on a sample of 840 children (5 to 16 years old) from ten small towns of the State of Bahia in northeastern Brazil. The objetive was to study, by using a cross sectional methodology, the evolution of schistosomiasis morbidity (hepatic and splenic enlargement) in children, and the role of the intensity of S. mansoni infection in this process. The children were analised in three age groups (5 to 8, to 12 and 13 to 16 years old) and classified as uninfected, mildly infected, moderately infected and heavily infected according to the number of eggs in the stool. In children aged 5 to 8 years, increasing egg counts were not associated with increasing frequencies of hepatic or splenic enlargement. In the 9 to 12 years old group and association was observed with the prevalence of hepatic enlargement, but not with the prevalence of spleen enlargement. In the oldest group, 13 to 16 years old, an association was observed with the prevalence of enlargement of both organs. It was evident that in this population schistosomiasis morbidity develops in the early period of life as a gradual process starting with liver enlargement and followed by spleen enlargement some years later. It was found that the intensity of infection has a fundamental role in this process, although there is a latent period of some years before clinical splenomegaly appears in moderate-heavily infected children. The Authors suggest that the prevalence of splenomegaly in the 13 to 16 years old group is a good measure of the community level of schistosomiasis morbidity and could be used to measure the impact of control programs.

O trabalho individual e o trabalho de grupo no processo de ensino-aprendizagem do estudo do meio no 1º Ciclo

Relatório Final apresentado à Escola Superior de Educação de Lisboa para obtenção de grau de mestre em Ensino do 1º e 2º ciclo do Ensino Básico

Spectroscopic Studies and Characterization of a Novel Electron-Transfer Chain

A novel two-component enzyme system from Escherichia coli involving a flavorubredoxin (FlRd) and its reductase was studied in terms of spectroscopic, redox, and biochemical properties of its constituents. FlRd contains one FMN and one rubredoxin (Rd) center per monomer. To assess the role of the Rd domain, FlRd and a truncated form lacking the Rd domain (FlRd¢Rd), were characterized. FlRd contains 2.9 ( 0.5 iron atoms/subunit, whereas FlRd¢Rd contains 2.1 ( 0.6 iron atoms/subunit. While for FlRd one iron atom corresponds to the Rd center, the other two irons, also present in FlRd¢Rd, are most probably due to a di-iron site. Redox titrations of FlRd using EPR and visible spectroscopies allowed us to determine that the Rd site has a reduction potential of -140 ( 15 mV, whereas the FMN undergoes reduction via a red-semiquinone, at -140 ( 15 mV (Flox/Flsq) and -180 ( 15 mV (Flsq/Flred), at pH 7.6. The Rd site has the lowest potential ever reported for a Rd center, which may be correlated with specific amino acid substitutions close to both cysteine clusters. The gene adjacent to that encoding FlRd was found to code for an FAD-containing protein, (flavo)rubredoxin reductase (FlRd-reductase), which is capable of mediating electron transfer from NADH to DesulfoVibrio gigas Rd as well as to E. coli FlRd. Furthermore, electron donation was found to proceed through the Rd domain of FlRd as the Rd-truncated protein does not react with FlRd-reductase. In vitro, this pathway links NADH oxidation with dioxygen reduction. The possible function of this chain is discussed considering the presence of FlRd homologues in all known genomes of anaerobes and facultative aerobes.

An ELISA method using serum derived HDAg for the sorological detection of HDV antigens and antibodies

One of the main difficulties related to the detection of the Hepatitis Delta Virus (HDV) antigen and antibody has been the source of the needed HD antigen since HDV containing human and animal livers are very difficult to obtain and since yield is low. This fact prompted us to try to use the serum of patients in the acute phase of HDV infection as a source of HDAg and turn to enzyme immunoassays (EIA) instead of RIA for the sake of easiness and economy in the amount of HDAg needed. The antigen for EIA was obtained from patients during the acute phase of HDV infection and the antibody from patients who have been carriers for many years. For the detection of the antigen, a sandwich type method was employed, whereas for the antibody a competition assay was developed. In order to assess the relative specificity and sensibility of the test, the antibody assay was compared to a commercial RIA (C. RIA, Abbott) and to a non-commercial RIA (NC RIA). Forty-two sera were tested by the two methods and only in two cases discrepant results were obtained. Its is concluded that: 1) sera from patients in the acute and chronic phases of HDV infection can be used as source of both antigen and antibody, for immunoassays; 2) EIA and RIA have comparable relative specificity and sensibility and 3) EIA is easier to perform, cheaper, non-hazardous, has a longer shelf-life and saves scarce HDAg.

Malária humana: padronização e optimização de testes sorológicos para diagnóstico individual e inquéritos soroepidemiológicos

O teste de imunofluorescência indireta (IFI) é considerado teste de referência na soroiogia da malária. Neste trabalho procuramos optimizar o teste empregando P. falciparum obtido de sangue humano e de cultura e P. vivax obtido de sangue de paciente como antígenos, para pesquisa de anticorpos IgG e IgM. Das variáveis técnicas estudadas melhores resultados foram obtidos quando os soros foram diluidos ern PBS contendo 1% de Tween 80 e as lâminas contendo a suspensão antigênica foram estabilizadas em dessecadores ou fixadas com acetona. Foi também padronizado o teste imunoenzimático ELISA com antigenos de P. falciparum obtidos em cultura. O estudo comparativo com o teste de imunofluorescência indireta para pesquisa de anticorpos IgG mostrou: a) nos pacientes primo infectados por P. falciparum a sensibilidade para ambos os testes foi de 71%; b) nos pacientes primo infectados pelo P. vivax a sensibilidade foi de 40% para ambos os testes; c) nos pacientes não primo infectados e com malária atual pelo P. falciparum a sensibilidade para ambos os testes foi de 100%; d) nos pacientes não primo infectados e com malária atual pelo P. vivax a sensibilidade foi de 85% para o teste ELISA e de 92% para a IFI; e) nos pacientes com malária mista a sensibilidade para ambos os testes foi de 100%. A especificidade da IFI foi de 100% e do teste ELISA 95% nos casos de indivíduos não maláricos. Os resultados obtidos sugerem ser o teste ELISA, uma boa alternativa para o teste de IFI, para a pesquisa de anticorpos IgG anti P. falciparum, na soroiogia da malária.

Rising stars: da ESTeSL para o mundo da radiologia

Serão apresentadas duas experiências que estudantes de radiologia da ESTeSL tiveram no estrangeiro: a experiência que as estudantes tiveram no seu ERASMUS em Viena de Áustria no 2º semestre de 2014/15 e o decorrer da Summer School OPTIMAX2015, em que as estudantes participaram em agosto de 2015, em Groningen, nos Países Baixos. Falar-se-á um pouco sobre os estudos em radiologia. Discutir-se-ão os desafios que são habitualmente apresentados a um estudante aquando da realização do seu artigo final de investigação, como se podem prevenir e ultrapassar esses obstáculos e construir um artigo científico de qualidade.

Exposição de profissionais de saúde a radiações ionizantes : confiança dos profissionais nas leituras de dosimetria individual

Resumo - O avanço tecnológico e científico no campo da Medicina tem favorecido a aplicação das radiações ionizantes nas áreas da Medicina Nuclear, Radiologia, Radioterapia e noutras especialidades relacionadas com a medicina de intervenção, como a Cardiologia. As Organizações de Saúde, com profissionais com risco de exposição a radiações ionizantes, são responsáveis, legislativamente, por assegurar a sua vigilância. Certificando a observação controlada das doses de exposição, os profissionais são monitorizados com dosímetros individuais e submetidos a vigilância médica. As leituras de dosímetria individual destes profissionais, originam por vezes, nos próprios, desconfianças e incertezas quanto aos valores medidos. Objectivo: avaliar o grau de confiança dos profissionais de saúde, expostos a radiações ionizantes, no sistema de monitorização individual. Metodologia: estudo observacional do tipo analítico- transversal, por questionário. Amostra de 190 Técnicos de Diagnóstico e Terapêutica de Cardiopneumologia, Medicina Nuclear, Radiologia, Radioterapia, expostos a radiações ionizantes, que exercem a sua actividade profissional em hospitais do concelho de Lisboa. Resultados: 51,1% dos Técnicos de Diagnóstico e Terapêutica não confiam nas leituras dos dosímetros, não se determinou uma relação estatisticamente significativa entre essa confiança, o tipo de dosímetro e a empresa que realiza as leituras. A confiança é maior nos que exercem em Organizações de Saúde públicas. 40,2% das Organizações de Saúde apresentam plano de vigilância médica, constatando-se uma falta de conformidade entre os Técnicos de Diagnóstico e Terapêutica, nalguns hospitais, relativamente à existência desse plano no respectivo hospital. A margem de erro associado ao estudo é de 5,09%, com um nível de confiança de 95%. -----------Abstract - The technological and scientific advances in the field of medicine has encouraged the application of ionizing radiation with a considerable positive contribution in the areas of Nuclear Medicine, Radiology, Radiotherapy and other specialties related to medical intervention like Cardiology. The Health Organizations that arise the risk of occupational exposure to ionizing radiation and is their responsibility to ensure monitoring, according the law. To ensure the monitoring of controlled doses of exposure, the exposed works use an individual dosimeter and undergo medical supervision. The individual dosimetry readings of health-care professionals, is sometimes questionable for themselves, arising distrust and uncertainty about the values measured. Objective: evaluate the degree of confidence of health professionals, exposed to radiation, on the system of individual monitoring. Methods: Observational study of cross-type analysis by questionnaire. Sample of 190 of technologists of Cardiopneumology, Nuclear Medicine, Radiology, Radiotherapy, exposed to ionizing radiation, which work in hospitals on the region of Lisbon. Results: 51.1% of technologists does not rely on the readings of their dosimeters, and wasn’t determined a statistically significant relationship between this trust, the type of dosimeter and the company that makes the readings(s) of their(s) dosimeter(s). The confidence in the readings is higher in public Health Organizations. 40.2% of the Health Organizations have medical monitoring plan and there is a disagreement between technologists, in some hospitals, relatively to the existence of this plan in their hospital. It was estimated that the u

Antibodies against non-structural c100/3 and structural core antigen of hepatitis C virus (HCV) in hemodialysis patients

Two groups of patients undergoing hemodialysis (HD) maintenance were evaluated for their antibody response to non-structural c100/3 protein and structural core protein of hepatitis C virus (HCV). Forty-six patients (Group 1) never presented liver abnormalities during HD treatment, while 52 patients (Group 2) had either current or prior liver enzyme elevations. Prevalence rates of 32.6% and 41.3% were found for anti-c100/3 and anti-HCV core antibodies, respectively, in patients with silent infections (Group 1). The rate of anti-c100/3 in patients of Group 2 was 71.15% and reached 86.5% for anti-HCV core antibodies. The recognition of anti-c100/3 and anti-core antibodies was significantly higher in Group 2 than in Group 1. A line immunoassay composed of structural and non-structural peptides was used as a confirmation assay. HBV infection, measured by the presence of anti-HBc antibodies, was observed in 39.8% of the patients. Six were HBsAg chronic carriers and 13 had naturally acquired anti-HBs antibodies. The duration of HD treatment was correlated with anti-HCV positivity. A high prevalence of 96.7% (Group 2) was found in patients who underwent more than 5 years of treatment. Our results suggest that anti-HCV core ELISA is more accurate for detecting HCV infection than anti-c100/3. Although the risk associated with the duration of HD treatment and blood transfusion was high, additional factors such as a significant non-transfusional spread of HCV seems to play a role as well. The identification of infective patients by more sensitive methods for HCV genome detection should help to control the transmission of HCV in the unit under study.