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- Análise evolutiva do perfil dos ex-fumadores e motivos de cessação.Publication . Leite, Andreia; Machado, Ausenda; Nunes, Baltazar; Matias Dias, CarlosAntecedentes/Objetivos: Em Portugal têm sido tomadas medidas para promover a cessação tabágica, nomeadamente a Lei do Tabaco (2008). Pretende-se analisar a evolução do perfil sociodemográfico dos ex-fumadores e os motivos da cessação. Métodos: Foram analisados os dados do painel ECOS (Em Casa Observamos Saúde). Este painel é constituído por cerca de 1000 Unidades de Alojamento (UA), contactáveis por telefone fixo ou móvel, estratificada por Região NUTII do Continente, com alocação homogénea. Em cada UA é inquirido um elemento com 18 ou mais anos. Os dados em análise foram recolhidos em Julho/2005, Junho/2008 e Outubro/ 2010. As estimativas, estratificadas por sexo, de prevalência de ex-fumadores e sua distribuição por características sociodemográficas, foram ponderadas para a população residente no Continente. Para tal utilizaram-se as estimativas oficiais da população residente e distribuição de UA por região (Instituto Nacional de Estatística). Resultados: Nos homens, as estimativas de prevalência de ex-fumadores nos 3 inquéritos, 2005, 2008 e 2010 foram 36,2% [Intervalo de Confiança a 95% (IC95): 30,6-41,9%], 43,8% (IC95: 36,1-51,5%) e 28,7% (IC95: 22,7-34,7%), respetivamente. Em 2005, os homens ex-fumadores, tinham maioritariamente mais de 65 anos (27,3% dos ex-fumadores) e ensino básico (49,2%). O principal motivo para cessação foram os problemas de saúde (49,6%; IC95: 40,1-59,2%). Este perfil não se alterou em 2008, tendo todos os ex-fumadores recentes (< 6 meses) referido que a lei não interferiu na sua decisão. Em 2010 a maioria dos ex-fumadores (93,0%; IC95: 88,0-97,8%) referiu ter cessado o consumo antes da implementação da lei e dos que o fizeram após a mesma, apenas 32,8% foram motivados pela lei. Nas mulheres verificaram-se prevalências de ex-fumadoras de 8,9% (IC95: 6,3-11,5%), 15,0% (IC95: 11,8-18,3%) e 13,2 (IC95: 9,6-16,8%), respetivamente. Em todos os inquéritos as ex-fumadoras tinham maioritariamente 35 a 44 anos (40,0% em 2005, 33,9% em 2008 e 39,2% em 2010) e habilitações literárias ao nível do ensino superior em 2005 e 2008 (46,9% e 36,8%) e do ensino básico em 2010 (48,8%) Em 2005, o principal motivo para ter deixado de fumar foram os problemas de saúde (22,7%; IC95: 10,5-34,5%). Igual resultado em 2008, contudo todas as ex-fumadoras recentes (há < 6 meses) referiram que a sua decisão foi influenciada pela lei. Em 2010, as ex-fumadoras referiram, ter cessado o consumo antes da publicação da lei (81,7%) e nenhuma referiu ter sido motivada pela mesma. Conclusiones: O perfil do ex-fumador não sofreu alterações substanciais ao longo do tempo, sendo o principal motivo para a cessação os problemas de saúde. As medidas futuras para a cessação tabágica deverão ter em conta as características dos grupos menos suscetíveis à mudança.
- The Nutritional Characterization of Rice Varieties Consumed in PortugalPublication . Mota, Carla; Santos, Mariana; Nascimento, Ana; Gueifão, Sandra; Coelho, Inês; Cabral, Margarida; Torres, Duarte; Castanheira, IsabelBackground and objectives: Portugal has the largest consumption of rice 15.8 kg/capita/year, in Europe. The present work aims to evaluate: rice nutritional profile, namely proximate, amino acid and mineral composition; arsenic content, as the most abundant contaminant and, compositional differences between varieties, year and place of harvest. Methods: Samples were collected from the most representative national rice producers. Analysis was carried out by following analytical methods: proximate by AOAC methods; amino acids by UPLC-DAD; starch, and amylase by enzymatic methods; mineral analysis by ICP-OES including copper, manganese, iron, zinc, magnesium, calcium, sodium, phosphorus and potassium and arsenic content by ICP-MS. Results: Analyzed rice showed higher amylopectin (54.7 ± 7.3 g/100g) content than amylose (33.5± 3.8 g/100g) and total protein content of 7.1 ± 0.3 g/100g a low content of fiber and fat (<1 g/100 g) was found. The most abundant essential amino acids were aromatic amino acids with a 7.5% of total protein. The most abundant minerals were potassium and phosphorus with levels ranging from 91 mg/100g to 107 mg/100g.The arsenic content was determined by ICP-MS and the values were below 600 µg/kg. Conclusions: Crop place showed to be the major source of variation in amino acid content. With respect to protein quality, cysteine, lysine, sulfur amino acids and isoleucine were considered limiting with protein digestibility corrected amino acids scores (PDCAAS) lower than 1. To assure the nutritional requirements for amino acids and subsequent protein synthesis, rice consumption must be followed by other food groups (e.g pulses, meat products) in order to provide the other essential amino acids. Rice contribution for mineral intake ranged from 3% DRI (iron and potassium) to 36 % DRI (manganese). In this study, a very weak correlation between arsenic levels and amino acid content was observed.
- Environment and Health in Children Day Care CentresPublication . Neuparth, Nuno; Papoila, Ana Luísa; Aelenei, Daniel; Cano, Manuela; Paixão, Paulo; Viegas, João; Martins, Pedro; Araújo Martins, José; Leiria Pinto, Paula; Caires, Iolanda; Pedro, Catarina; Nogueira, Susana; Mendes, Ana; Aguiar, Fátima; Teixeira, João Paulo; Proença, Carmo; Piedade, Cátia; Santos, Madalena; Silvestre, Maria José; Brum, Laura; Nunes, Baltazar; Guiomar, Raquel; Curran, Martin D.; Carvalho, Ana; Marques, Teresa; Virella, Daniel; Alves, Marta; Marques, João; Rosado-Pinto, José; Neuparth, Nuno; Aelenei,Daniel; Caires, Iolanda; Teixeira, João Paulo; Viegas, João; Cano, Manuela; Pinto, Paula LeiriaThis project addresses a set of common clinical problems in the context of children attending day care centres. It is common sense that children get sick more often as soon as they start attending a day care centre on a daily basis and this is particularly true for some groups at risk, as wheezing infants and wheezing pre-school children. Concerning this, some questions remain unclear: 1. The role of indoor air quality - what is the health impact of indoor air environment (including indoor pollutants, house dust mite, temperature and humidity) in wheezing children? 2. The role of virus infections - We don’t know the real role of virus infections in respiratory conditions at day care centre level. There is a lack of information concerning how indoor air environment influence virus infections. 3. The role of building ventilation - what is the impact of building ventilation in the health of wheezing and non-wheezing children? How is ventilation affecting indoor air quality? How is ventilation of day care centres affected by the structure of the buildings? 4. Social impact of this study - what should be the recommendations to improve IAQ? What is new in this project is the collaboration of a health team (medical doctors and other health professionals) with environment specialists, mechanical, civil engineers, epidemiologists and statisticians.
- Complex III deficiency in a Portuguese family: expanding the clinical phenotypePublication . Nogueira, Célia; Nesti, Claudia; Meschini, M. Clara; Carrozzo, Rosalba; Barros, José; Sá, Maria José; Azevedo, Luisa; Santorelli, Filippo; Vilarinho, Laura
- Biochemical data as important clues for diagnosis of SUCLA2 defectsPublication . Nogueira, Célia; Garcia, Paula; Diogo, Luisa; Valongo, Carla; Santorelli, Filippo; Vilarinho, Laura
- Antiviral resistance: influenza BPublication . Conde, Patrícia; Guiomar, Raquel; Cristóvão, Paula; Pechirra, PedroCurrently circulating influenza viruses are resistant to adamantanes and except for a low number of sporadic cases most are sensitive to neuraminidase inhibitors (NI). Adamantanes are ineffective against influenza B viruses and although NI-resistant influenza B viruses have been rarely reported, recently in the United States was identified one cluster of influenza B viruses with reduced susceptibility to NI and with the I221V substitution in the active site of the neuraminidase. Despite the low prevalence of oseltamivir-resistant influenza viruses, the constant evolution of influenza requires the monitoring of antiviral resistance among these viruses in the community. This is very important for the clinical management of severe influenza cases as to the detection of novel genetic markers associated with antiviral resistance. This study reports the antiviral susceptibility to neuraminidase inhibitors of influenza B viruses isolated in Portugal during the 2010/2011, 2011/2011 and 2012/2013 seasons. Over the period of 3 influenza seasons, 146 influenza B viral strains were selected for phenotypic fluorescent assays in order to assess their susceptibility to NI, oseltamivir and zanamivir. For this purpose, was determined the NI concentration required to inhibit 50% of each influenza virus neuraminidase activity (IC50). The IC50 baseline of influenza B viruses was calculated for both oseltamivir and zanamivir using the Robust Excel programme. The neuraminidase gene segments were also monitored for the presence of the main molecular markers, associated with the resistance to neuraminidase inhibitors in influenza B viruses. All analysed influenza B strains proved to be susceptible to oseltamivir and zanamivir. In the 2010/2011 season the determined IC50 values ranged from 0 to 70 nM for oseltamivir and from 0 to 11nM to zanamivir. The zanamivir IC50 median value was about 8-fold lower than oseltamivir IC50 median value. Statistical analysis revealed the presence of one outlier (B/Lisboa/13/2010, 71.08nM) for oseltamivir (2-fold reduction in susceptibility) and four minor outliers (B/Lisboa/15/2010: 9.52 nM; B/Lisboa/19/2010: 9.25 nM; B/Lisboa/51/2010: 10.94 nM and B/Lisboa/53/2010: 9.62 nM) for zanamivir (3-fold reduction in susceptibility) comparing to the median IC50 value. During the 2011/2012 season IC50 values ranged from 23.0 to 38.38 nM (oseltamivir) and from 2.97 to 9.07nM (zanamivir). This season were not found minor or major outliers. The IC50 values obtained in 2012/2013 ranged from 7.91 to 84.84 nM (oseltamivir) and from 1.48 to 5.88 nM (zanamivir). During this last season were found 6 minor and 3 major outliers for oseltamivir and 13 minor outliers for zanamivir. Along the three seasons, the median IC50 values for both NI were higher among the B/Victoria than B/Yamagata viruses. None of the actually known mutations associated with resistance of influenza B viruses to NI was found in the neuraminidase gene (R150K, D197E/N/Y, I221T/V, N294S, R374K and G407S). Influenza B viruses isolated in Portugal during the last three seasons were susceptible to the neuraminidase inhibitors. Portuguese influenza B strains revealed higher susceptibility to zanamivir than to oseltamivir, as observed in other countries. The oseltamivir IC50 values were also different between viruses from the B/Victoria and B/Yamagata lineages, however this was not statistically demonstrated due to the small number of analysed viruses. Among B/Yamagata strains the oseltamivir and zanamivir IC50 values were higher in 2011/2012 than in 2010/2011 and 2012/2013 seasons.
- Severe acute respiratory infections in the 2012/2013 season studied by the Portuguese Laboratory Network for Influenza DiagnosisPublication . Guiomar, Raquel; Pechirra, Pedro; Conde, Patrícia; Cristóvão, Paula; Silvestre, Maria José; Almeida Santos, Madalena; Sobrinho Simões, Joana; Costa, Maria do Rosário; Amaral, Susana; Guimarães, João Tiago; Ribeiro, Graça; Correia, Lurdes; Fernandes, Aida; Milho, Luís; Fernandes, Paula Luísa; Andrade, Graça; Mota Vieira, Luísa; Cabral, Rita; Moniz, Raquel; Pereirinha, Tania; Bruges Armas, Jacome; Pimentel Couto, Ana Rita; Soares, Marta; Melo Cristino, José; Carvalho, Dinah; Ribeiro, Carlos; Barreto, Rosário; Côrte-Real, Rita; Branquinho, Paula; Ramos, Maria Helena; Castro, Ana Paula; Caldeira, Filomena; Maurílio, Manuel; Cunha, Mário; Ornelas, Carmo; Almeida, SofiaDuring the 2009/10 influenza pandemic, a network of 14 laboratories located in the main reference hospitals from Portugal mainland, Madeira and Azores was established for the diagnosis of the new influenza A(H1N1)2009 pandemic strain. Since then, the network performs laboratory diagnosis of influenza as well as other respiratory pathogens, thus contributing to the laboratory diagnosis of respiratory disease in Portugal. This network is a valuable complement of the National Influenza Surveillance Programme (mainly based on primary healthcare units), enabling a more accurate knowledge of the aetiology of the severe respiratory infections, especially in hospitalized cases. The present study describes the severe acute respiratory infections, in the 2012/2013 season, diagnosed by the laboratory network. From the 14 laboratories, 11 reported cases of respiratory disease during 2012/2013 season. The laboratory network performs diagnosis of influenza A and B viruses and other respiratory agents by PCR based methods, also enabling the detection of mixed infections. All 14 laboratories perform the detection of influenza A(H1)pdm09, 4 perform the influenza A(H1) seasonal and A(H3) subtyping, and 10 participants also detect influenza B. Eight laboratories implemented methodologies for the detection of other infectious agents associated with respiratory disease. The antigenic characterization of 8 isolated viruses [3 A(H1)pdm09 and 5 B/Yamagata] was performed at the National Influenza Reference Laboratory. The genetic analysis of the HA1 subunit of the haemagglutinin gene was performed in 17 viruses [7 A(H1)pdm09, 1 A(H3) and 9 B/Yamagata]. Twenty nine A(H1)pdm09 and 5 B/Yamagata were tested for antiviral susceptibility [PCR(NA)-H275Y and/or MUNANA phenotypic assays for oseltamivir and zanamivir]. The 11 laboratories reported a total of 1470 respiratory disease cases, from week 39/2012 to 21/2013 [peak of 205 (13.9%) cases during week 10/2013]. Influenza was identified in 504 cases. Influenza A was detected in 352 (70.0%) cases: 297 (59.0%) cases were A(H1)pdm09, 48 (10.0%) cases were not subtyped, and 7 (1.0%) cases were A(H3). Influenza B was identified in 152 (30%) of the influenza cases. During the 2012/2013 season, 311 (21.2%) reported cases were hospitalized in intensive care units (ICU), the majority of them had between 50-54 years (34; 10.9%), followed by the age groups 45-49 and 55-59 years old (28; 9.0% each). The causal agent was identified in 160 (51.4%) ICU cases. Influenza was identified in 120 (38.5%) patients, other respiratory agents were detected in 40 (12.8%), within these, multiple infections were present in 18 (5.7%). Bacteria were identified in 31 (10.0%) cases mainly associated with RSV and hRV. Among ICU influenza cases, the most detected virus was A(H1)pdm09 (82; 62.0%). However, cases of A(H3) (3; 2.0%), A unsubtyped (8; 7.0%) and B (27; 23.0%) were also detected. As expected, the highest number of ICU influenza positive cases was detected in week 10/2013 (18; 15.0%), coincident with the highest number of influenza cases during all season. ICU flu cases were detected predominantly in individuals between 50-54 years (18; 15.0%). From the ICU reported cases, 6 (1.9%) died. The influenza A(H1)pdm09 virus was detected in 2 man between 50-59 years old from these 6 fatal outcomes. The isolated influenza A viruses were similar to the 2012/2013 vaccine strains. The influenza B/Yamagata viruses showed a greater antigenic and genetic variability. The Portuguese Laboratory Network for Influenza Diagnosis plays a major role in the diagnosis of acute respiratory infections in Portugal, providing a more accurate knowledge of the respiratory agents involved. During the 2012/2013 season, the influenza A(H1)pdm09 virus predominated in co-circulation with influenza B virus. The A(H1)pdm09 virus was the responsible for the majority of the flu cases admitted in the ICU and may have been the cause of death in two cases. Bacterial and other viral agents have been identified in some of the severe cases reported. The majority of the characterized influenza viruses were similar to the vaccine strains and none of the virus showed reduced susceptibility to oseltamivir or zanamivir.
- Annexin A11 gene polymorphism (R230C variant) and sarcoidosis in a Portuguese populationPublication . Morais, A.; Lima, B.; Peixoto, M.; Melo, N.; Alves, H.; Marques, J.A.; Delgado, L.A recent genome-wide association study detected a protective effect for the annexin A11 rs1049550*T allele (R230Cvariant) in susceptibility to sarcoidosis. We evaluated the association between rs1049550 C/T and sarcoidosis susceptibility, distinct disease phenotypes and evolution in a Portuguese population. We performed a case-control study of 208 patients and 197 healthy controls. Samples were genotyped for rs1049550 C/T using real-time polymerase chain reaction. The frequency of the annexin A11 rs1049550*T allele was significantly lower in patients than in controls (33.2 vs 44.9%, P < 0.001). Odds ratio of 0.52 and 0.44 were obtained, respectively for carriers of one (CT) and two (TT) copies normalized to the CC wild-type genotype (P < 0.001). There were no significant differences in patients with and without Löfgren syndrome. A significant increase in the frequency of the T allele was observed in patients with bronchoalveolar lavage (BAL) fluid neutrophilia (P = 0.04). No significant associations were seen for lung function pattern, radiological stages or different forms of disease evolution. Our study confirms that rs1049550*T allele exerts a significant protective effect on sarcoidosis susceptibility. Given the role of annexin A11 in cell division, apoptosis and neutrophil function, this polymorphism may affect key elements of granulomatous and interstitial inflammation in sarcoidosis.
- Microbiological and mycological beach sand quality in a volcanic environment: Madeira archipelago, PortugalPublication . Pereira, Elisabete; Figueira, Celso; Aguiar, Nuno; Vasconcelos, Rita; Vasconcelos, Sílvia; Calado, Graça; Brandão, João; Prada, SusanaMadeira forms a mid-Atlantic volcanic archipelago, whose economy is largely dependent on tourism. There, one can encounter different types of sand beach: natural basaltic, natural calcareous and artificial calcareous. Microbiological and mycological quality of the sand was analyzed in two different years. Bacterial indicators were detected in higher number in 2010 (36.7% of the samples) than in 2011 (9.1%). Mycological indicators were detected in a similar percentage of samples in 2010 (68.3%) and 2011 (75%), even though the total number of colonies detected in 2010 was much higher (827 in 41 samples) than in 2011 (427 in 66 samples). Enterococci and potentially pathogenic and allergenic fungi (particularly Penicillium sp.) were the most common indicators detected in both years. Candida sp. yeast was also commonly detected in the samples. The analysis of the 3rd quartile and maximum numbers of all indicators in samples showed that artificial beaches tend to be more contaminated than the natural ones. However, a significant difference between the variables was lacking. More monitoring data (number of bathers, sea birds, radiation intensity variation, and a greater number of samples) should be collected in order to confirm if these differences are significant. In general, the sand quality in the archipelago's beaches was good. As the sand may be a vector of diseases, an international common set of indicators and values and a compatible methodologies for assessing sand contamination, should be defined, in order to provide the bather's with an indication of beach sand quality, rather than only the water.
- Comparison between high-performance and ultra-high performance liquid chromatography methods for cholesterol determination in foodsPublication . Costa, H.S.; Oliveira, M.B.; Sanches-Silva, A.; Albuquerque, T.G.Background and Objectives: Gas chromatography is still the most widely used technique for cholesterol determination, however liquid chromatography was shown to provide the most sensitive and accurate results. The purpose of this study was to compare high-performance and ultra-high performance liquid chromatography methods (HPLC and UHPLC) for cholesterol determination in foods. Methods: For HPLC method, a Supelcosil LC-18-DB (15 cm x 4.6 mm x 3.0 µm) column was used, whereas in UHPLC it was an Acquity BEH C18 (2.1 mm x 50 mm x 1.7 µm). For both methods, a UV detector (210 nm) and a mobile phase (acetonitrile/isopropanol, 70:30, v/v) were used. Methods were validated with respect to: linearity, limit of detection (LOD), limit of quantification (LOQ), recovery, accuracy and precision. Results: Both methods were suitable for cholesterol determination in foods, with linearity ranging from 0.005 to 0.40 mg/mL (HPLC - r2 = 0.9991; UHPLC – r2 =0.9992). However with UHPLC method, a lower LOQ (2.4 µg/mL) and LOD (0.73 µg/mL) and best recovery rates were achieved. Moreover, run time decreases from 4.5 min to 2 min. Also, it was possible to significantly reduce the amount of solvents used, since for UHPLC the flow rate is 0.3 mL/min instead of 1.2 mL/min. Conclusions: In this work, a new UHPLC method has been developed for quantification of cholesterol in foodstuffs. HPLC and UHPLC analysis have been compared. Analysis time, solvent consumption, and analysis cost are very important in many analytical laboratories. Overall, UHPLC can offer significant improvements compared with conventional HPLC.