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- Indicadores de Prognóstico da Carcinogénese do Colo do Útero Associada à Infeção por Vírus do Papiloma HumanoPublication . Oliveira, Ana; Delgado, Candida; Verdasca, Nuno; Pista, Angela[PT] Introdução/Objetivos: A infeção persistente pelo Vírus do Papiloma Humano de alto risco (HPVar) é considerada como a causa necessária, embora não suficiente, para o desenvolvimento do cancro do colo do útero, sugerindo que outros fatores estarão envolvidos no processo de carcinogénese. Este estudo pretendeu avaliar indicadores de prognóstico da persistência da infeção por HPV, nomeadamente o estado físico e a carga viral dos HPV 16 e 18 e a superexpressão dos transcritos do RNAm dos HPV 16, 18, 31, 33 e 45, num grupo de mulheres com ou sem sintomatologia clínica e citopatológica. Material e Métodos: Foram estudadas 378 alíquotas de células epiteliais congeladas pertencentes a utentes dos centros de saúde do Serviço Nacional de Saúde e de clínicas privadas, referenciadas para teste HPV, entre Janeiro de 2007 e Dezembro de 2010. De acordo com o diagnóstico citopatológico, foram definidos cinco grupos: normal, ASCUS, LSIL, HSIL e carcinoma invasivo do colo do útero. Para a determinação do estado físico do DNA e da carga viral dos HPV 16 e 18 foi utilizada metodologia de PCR em tempo real, e para a superexpressão dos transcritos dos oncogenes E6 e E7 o sistema comercial NucliSENS EasyQ HPV. Os indicadores foram analisados em associação com os tipos de lesão do colo do útero. Para a análise estatística foi utilizado o o programa informático SPSS versão 16.0 e o teste de Chi-Quadrado. Resultados: Os resultados mostraram ausência de associação estatisticamente significativa entre a gravidade da lesão e o estado físico do DNA dos HPV 16 e 18. A superexpressão dos transcritos do RNAm E6/E7 e a carga viral dos HPV 16 e 18 aumentaram significativamente em função do grau da lesão. Conclusões: Os resultados obtidos sugerem que a determinação do estado físico do DNA dos HPV 16 e 18, isoladamente, não constitui um indicador de prognóstico para o desenvolvimento e progressão das lesões. A superexpressão dos transcritos dos oncogenes E6 e E7 está associada à progressão das lesões do colo do útero e apresenta maior especificidade no diagnóstico precoce das lesões pré-malignas. A quantificação do DNA dos HPVar pode ser um indicador promissor de prognóstico das lesões pré-neoplásicas do colo do útero.
- Human papillomavirus type distribution in cervical intraepithelial neoplasia grade 2/3 and cervical cancer in Portugal: a CLEOPATRE II StudyPublication . Pista, Angela; de Oliveira, Carlos F.; Lopes, Carlos; Cunha, Maria João; CLEOPATRE Portugal Study GroupaOBJECTIVE: Cervical cancer is the third most frequent cancer in women, worldwide and etiologically associated with infection by human papillomavirus (HPV). Following the results of the first epidemiologic population-based CLEOPATRE study in Portugal, it was important to understand the HPV type-specific distribution in women with cervical intraepithelial neoplasia (CIN) grades 2 and 3 and invasive cervical cancer (ICC). METHODS: This was an observational, multicenter, cross-sectional study with retrospective data collection. Between January 2008 and May 2009, paraffin-embedded samples of histologically confirmed cases of CIN2, CIN3, and ICC were collected from the 5 regional health administrations in mainland Portugal. Eligible samples were sent to 2 central laboratories for histological reassessment and HPV genotyping. Prevalence estimates were calculated together with 95% confidence intervals. RESULTS: A total of 582 samples, 177 cases of CIN2, 341 of CIN3, and 64 of ICC, were included. The mean age of participants was 41.8 years (range, 20-88 years). The overall HPV prevalence was 97.9% with a higher prevalence of high-risk genotypes, particularly HPV 16. Multiple infections were observed in 11.2% of the cases. Human papillomavirus prevalence was 95.5% in CIN2, 99.4% in CIN3, and 96.9% in ICC. The 8 more frequent genotypes in order of decreasing frequency were HPV 16, 31, 58, 33, 51, 52, 18, and 35 in CIN2 and HPV 16, 31, 33, 58, 52, 35, 18, and 51 in CIN3. In ICC cases, the 12 detected HPV genotypes were HPV 16, 18, 31, 33, 45, 51, 52, 53, 56, 58, 59, and 73. However, HPV 53 and 73 were always associated to other high-risk genotypes. Human papillomavirus types 31, 51, 52, 56, and 59 were detected in 1 case each. CONCLUSIONS: Human papillomavirus prevalence and patterns of type-specific HPV positivity were comparable with other studies. Current HPV vaccines should protect against HPV genotypes responsible for 77.4% of ICC in Portugal.
- Effects of occupational exposure to tobacco smoke: is there a link between environmental exposure and disease?Publication . Pacheco, Solange A.; Torres, Vukosava M.; Louro, Henriqueta; Gomes, Filomena; Lopes, Carlos; Marçal, Nelson; Fragoso, Elsa; Martins, Carla; Oliveira, Cátia L.; Hagenfeldt, Manuela; Bugalho-Almeida, António; Penque, Deborah; Simões, TâniaIn a previous study, evidence was provided that indoor secondhand tobacco smoke (SHS) air pollution remains high in Lisbon restaurants where smoking is allowed, regardless of the protective measures used. The aim of this study was to determine in these locations the levels of polycyclic aromatic hydrocarbons (PAH) associated with the particulate phase of SHS (PPAH), a fraction that contains recognized carginogens, such as benzo[a]pyrene (BaP). Data showed that restaurant smoking areas might contain PPAH levels as high as 110 ng/m(3), a value significantly higher than that estimated for nonsmoking areas (30 ng/m(3)) or smoke-free restaurants (22 ng/m(3)). The effective exposure to SHS components in restaurant smoking rooms was confirmed as cotinine levels found in workers' urine. Considering that all workers exhibited normal lung function, eventual molecular changes in blood that might be associated with occupational exposure to SHS and SHS-associated PPAH were investigated by measurement of two oxidative markers, total antioxidant status (TAS) and 8-hydroxyguanosine (8-OHdG) in plasma and serum, respectively. SHS-exposed workers exhibited higher mean levels of serum 8-OHdG than nonexposed workers, regardless of smoking status. By using a proteomics approach based on 2D-DIGE-MS, it was possible to identify nine differentially expressed proteins in the plasma of SHS-exposed nonsmoker workers. Two acute-phase inflammation proteins, ceruloplasmin and inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), were predominant. These two proteins presented a high number of isoforms modulated by SHS exposure with the high-molecular-weight (high-MW) isoforms decreased in abundance while low-MW isoforms were increased in abundance. Whether these expression profiles are due to (1) a specific proteolytic cleavage, (2) a change on protein stability, or (3) alterations on post-translational modification pattern of these proteins remains to be investigated. Considering that these events seem to precede the first symptoms of tobacco-related diseases, our findings might contribute to elucidation of early SHS-induced pathogenic mechanisms and constitute a useful tool for monitoring the effects of SHS on occupationally exposed individuals such as those working in the hospitality industry.
- Deliverable 6: Characterisation of manufactured nanomaterials for their clastogenic/aneugenic effects or DNA damage potentials and correlation analysis. Final report.Publication . NANOGENOTOX Joint Action; WP6 partners; Fessard, V.Objectives: To generate data from selected in vivo genotoxicity tests, and to assess the correlation between in vivo and in vitro results taking into account the kinetic results. Schedule: The WP6 lasted for 16 months from October 2011 until the end of the Joint Action (February 2013). Few months at the beginning were devoted to trainings and trials before performing the experiments on Manufactured Nanomaterials (MNs). Even though, some last results are not yet available and will be collected after the end of the Joint Action, especially on the micronucleus on colon as well as on histology.
- Fatty acids composition and total fat content in traditional foods from Black Sea Area countriesPublication . Albuquerque, T.G.; Sanches-Silva, A.; Finglas, P.; D'Antuono, L.F.; Trichopoulou, A.; Alexieva, I.; Boyko, N.; Costea, C.; Fedosova, K.; Karpenko, D.; Kilasonia, Z.; Koçaoglu, B.; Costa, H.S.Objectives: This work was performed within the collaborative research program Sustainable exploitation of bioactive components from the Black Sea Area traditional foods (BaSeFood), funded by the European Commission. The overall objective of this study was to determine the total fat content and fatty acids (FA) composition among 33 traditional foods from six Black Sea Area countries (BSAC). Method: Total fat analysis was performed by an acid hydrolysis method with Soxhlet extraction (AOAC 948.15, 2000). Preparation of FA methyl esters was carried out by a combined method of methylation and transesterification. For FA determination chromatographic analysis was performed using a gas chromatograph with mass detector. FA methyl esters were separated in an Agilent HP-88 column (100 m x 0.25 mm i.d., 0.25 µm). Helium was the carrier gas. Split ratio of 50:1 and an injection volume of 1.0 µL were used. FA methyl esters were identified by comparing the retention times of the standards with those of the samples. Results: Total fat content ranged between 0.138 ± 0.0 g/100 g for Uzvar (Ukraine) and 99.9 ± 0.0 g/100 g for flax oil and mustard oil. From the 33 analysed traditional foods, 27.3% had fat contents below the limit of quantification (< 0.1 g/100 g). The highest content for saturated and polyunsaturated FA was found for flax oil (15.3 ± 0.6 g/100 g and 58.5 ± 0.5 g/100 g, respectively). Mustard oil showed the highest value for monounsaturated FA (55.1 ± 0.7 g/100 g), although it also has a high polyunsaturated FA content (38.0 ± 0.6 g/100 g of edible portion). Conclusions: In general, there was a great variability of results with respect to the FA profile of the analyzed traditional foods from BSAC, since there is a wide range of traditional foods from different countries and cultures. Some traditional foods from BSAC are a good source of unsaturated FA which are related to several health benefits. Acknowledgements: The research leading to these results has received funding from the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n.º 227118.
- Identifying Needs of Food Composition Database Users – PortFIR experiencePublication . Dias, M. Graça; Viegas, Silvia; Brazão, Roberto; Oliveira, LuisaInformation sharing and management among all the intervenients in food chain is crucial to improve food safety and to promote and protect public health. It optimizes resources usage and provides evidence support to define and promote policies in food and nutrition and to increase the efficiency of food productive systems. Objectives. To share information among Portuguese Food Composition Table users, to identify their needs, in order to define priorities for the update of the National Food Composition Table. Material and Methods. INSA, IP established a partnership with GS1 Portugal, to develop a programme, “Portuguese Food Information Resource” (PortFIR), to implement national networks of knowledge sharing on nutrition and food safety. The program has the collaboration of governmental and private organizations encompassing health, agriculture and economy sectors and includes a food information web portal integrated in a Food Information Network Management System (SGRIA). Two networks were created to date, “Food Composition” and “Food Microbiological Information”, acting through working groups (WG), some of which are transversal to both networks. “Users” WG (GTU) developed a questionnaire to evaluate the needs and satisfaction of the users of the Portuguese Food Composition Table (FCT). The questionnaire was implemented on-line from July to October 2011 using the software LimeSurvey®, and FCT users were invited to participate through direct contact by email and/or through professional associations and networks. Results. The analysis of the responses revealed that the preferred format of the FCT is “on-line” (52%) (options: on-line, CD-Rom, book, other) and that 80% of responding users are satisfied with either FCT format. Regarding additional information needs, users refer allergens (50%), food portion (39%), fatty acids n-3 and n-6 (25%), and traditional dishes (23%). Respondents consider the number of foods, micronutrients and composite dishes as the main limitations of the present FCT. Conclusions. The cooperation between data producers, users and compilers is contributing to optimize the usage of National resources and to disseminate the knowledge about nutrition and food safety within the country. The information obtained through “PortFIR – GTU” will allow compilers to establish informed priorities taking into account the expressed needs of users across the food chain.
- Volatile organic compounds in day care centers constitute a risk factor for absence due to weezingPublication . Carreiro-Martins, Pedro; Papoila, Ana Luísa; Alves, Marta; Caires, Iolanda; Pedro, Catarina; Manilha, Maria do Carmo; Cano, Maria Manuela; Teixeira, João Paulo; Virella, Daniel; Leiria-Pinto, Paula; Rosado-Pinto, José; Neuparth, Nuno
- MPV17: fatal hepatocerebral presentationPublication . Nogueira, Célia; Souza, Carolina; Husny, Antonette; Derks, Terry; Santorelli, Filippo; Vilarinho, Laura
- Antioxidant activity of Annona cherimola Mill. (Annonaceae)Publication . Albuquerque, T.G.; Santos, F.; Sanches-Silva, A.; Costa, D.; Costa, H.S.Objectives: In this study, the antioxidant capacity of pulp and seeds of ripe fruits of Annona cherimola Mill. was determined. Moreover, two organic solvents were compared regarding their radical scavenging activity (RSA). The present research intends to carry out a study on the best extraction method for further use in the evaluation of RSA in several cultivars of traditional fruits from Madeira’s island. Method: Annona cherimola Mill. fruits were acquired in local supermarkets. Fruits were selected and peel, seeds and pulp were separated. Then, each part of the fruits was homogenised in a blender for 1 min at 5000 rpm. Samples (pulp or seeds) were extracted with 25 mL of solvent (ethanol 90%, v/v, or methanol) and filtered through cotton wool. Afterwards, DPPH radical was used for antioxidant capacity determination, which in presence of an antioxidant is reduced to DPPH-H decreasing its spectrophotometric absorbance at 517 nm. The scavenging activity of the extracts was expressed as the percentage of inhibition of the DPPH radical. Results: The IC50 value, representing the concentration of the extract that gives rise to 50% reduction in DPPH absorbance was determined by linear regression analysis. IC50 for pulp and seeds was lower for ethanolic extracts than for methanolic extracts. Conclusions: Due to the importance of antioxidant compounds to prevent several diseases such as cancer, and cardiovascular diseases, it is very important to compare the antioxidant capacity of different fruits, fruit components, as well as extract solvents. Ethanol gave higher IC50 in all the tested extracts. These results will support further research on traditional fruits from Madeira’s island. Acknowledgements: Tânia Gonçalves Albuquerque is grateful for research grant (BRJ/DAN-2012) funded by INSA. Ana Sanches Silva is grateful for the postdoctoral contract in the frame of the Program ‘Science 2007’, funded by FCT.
- Human Papillomavirus Type Distribution in Cervical Intraepithelial Neoplasia Grade 2/3 and Cervical Cancer in Portugal: a CLEOPATRE II StudyPublication . Pista, A.; Oliveira, C.F.; Lopes, C.; Cunha, M.J.; CLEOPATRE Portugal Study GroupaObjective: Cervical cancer is the third most frequent cancer in women, worldwide, and etiologically associated with infection by human papillomavirus (HPV). Following the results of the first epidemiologic population-based CLEOPATRE study in Portugal, it was important to understand the HPV type-specific distribution in women with cervical intraepithelial neoplasia (CIN) grade 2 and 3 and invasive cervical cancer (ICC). Methods/materials: Observational, multicenter, cross-sectional study with retrospective data collection. Between January 2008 and May 2009, paraffin-embedded samples of histologically confirmed cases of CIN2, CIN3 and ICC were collected from the 5 regional health administrations in mainland Portugal. Eligible samples were sent to two central laboratories for histological reassessment and HPV genotyping. Prevalence estimates were calculated together with 95% confidence intervals. Results: A total of 582 samples, 177 cases of CIN2, 341 of CIN3 and 64 of ICC, were included. The mean age of participants was 41.8 years (20-88 years). The overall HPV prevalence was 97.9% with a higher prevalence of high-risk genotypes, particularly HPV 16. Multiple infections were observed in 11.2% of the cases. HPV prevalence was 95.5% in CIN2, 99.4% in CIN3 and 96.9% in ICC. The eight more frequent genotypes in order of decreasing frequency were HPV 16, 31, 58, 33, 51, 52, 18, and 35 in CIN2 and HPV 16, 31, 33, 58, 52, 35, 18, and 51 in CIN3. In ICC cases, the detected HPV genotypes were HPV 16, 18, 31, 33, 45, 51, 52, 53, 56, 58, 59 and 73. However, HPV 53 and 73 were always associated to other high-risk genotypes. HPV 31, 51, 52, 56, and 59 were detected in one case each. Conclusions: HPV prevalence and patterns of type-specific HPV positivity were comparable with other studies. Current HPV vaccines should protect against HPV genotypes responsible for 77.4% of ICC in Portugal.