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- Helicobacter pullorum Isolated from Fresh Chicken Meat: Antibiotic Resistance and Genomic Traits of an Emerging Foodborne PathogenPublication . Borges, Vítor; Santos, Andrea; Correia, Cristina Belo; Saraiva, Margarida; Ménard, Armelle; Vieira, Luís; Sampaio, Daniel A.; Pinheiro, Miguel; Gomes, João Paulo; Oleastro, MónicaMeat and meat products are important sources of human intestinal infections. We report the isolation of Helicobacter pullorum strains from chicken meat. Bacteria were isolated from 4 of the 17 analyzed fresh chicken meat samples, using a membrane filter method. MIC determination revealed that the four strains showed acquired resistance to ciprofloxacin; one was also resistant to erythromycin, and another one was resistant to tetracycline. Whole-genome sequencing of the four strains and comparative genomics revealed important genetic traits within the H. pullorum species, such as 18 highly polymorphic genes (including a putative new cytotoxin gene), plasmids, prophages, and a complete type VI secretion system (T6SS). The T6SS was found in three out of the four isolates, suggesting that it may play a role in H. pullorum pathogenicity and diversity. This study suggests that the emerging pathogen H. pullorum can be transmitted to humans by chicken meat consumption/contact and constitutes an important contribution toward a better knowledge of the genetic diversity within the H. pullorum species. In addition, some genetic traits found in the four strains provide relevant clues to how this species may promote adaptation and virulence.
- Os ovos e o NatalPublication . Viegas, Silvia; Fernandes, Paulo; Coelho, MarianaDivulgação sobre o consumo dos ovos no âmbito do Natal, na perspectiva de segurança alimentar.
- Analysis of the translatome by ribosome profiling in colorectal cancerPublication . Silva, Joana; Romão, Luísa; Luchessi, AugustoColorectal cancer (CRC) is a serious health problem due to its high incidence and mortality rates despite the major advances in cancer therapeutic approaches [1]. CRC carcinogenesis progression is based in a continuous accumulation of genetic alterations that leads to variations in the overall gene expression profiles [2]. This creates the need for deep analysis of cancer gene expression patterns and, thus, a more reliable understanding of the human proteome to disclose the molecular and cellular pathways as well as the regulatory mechanisms involved in cancer progression [2-4]. Genome wide analyses of gene expression have so far focused on the abundance of mRNA species as measured either by microarray or, more recently, by RNA deep sequencing [5,6]. However, neither approach provides information on protein synthesis, an important end point of gene expression [5,7]. Ribosome profiling is an emerging technique that uses deep sequencing to monitor in vivo translation and provide global and quantitative measurements of translation [7,8]. It can also reveal unexpected complexity in translation, including the presence of ribosomes outside of classical protein-coding regions of the transcriptome [5]. In this approach, translation is profiled by nuclease footprinting of ribosomes on RNA templates and high-throughput sequencing in order to determine the precise positions of ribosomes on a transcript and its overall density [8]. Ribosome profiling studies have been performed in cancer cell lines, where they showed an increase in overall protein identification and new proteins not yet annotated that possibly were originated from N-terminal extensions or upstream open reading frames (uORFs) [9-12]. The main goal of this project is to determine the changes between the translatome of CRC and normal colorectal cells and the role of such alterations in the tumorigenesis process of CRC cells. For this purpose, we will perform ribosome profiling in normal (NCM460) and CRC (HCT116) cell lines. Bioinformatics and gene ontology analysis of the translated mRNAs will elucidate the main cellular pathways through which the corresponding proteins are involved in CRC progression. Then, we will dissect which of these proteins can interfere and induce cell survival of CRC cells. Furthermore, we aim to analyze the potential contribution of translatable short alternative ORFs (AltORFs) and/or the corresponding peptides towards CRC progression. This information will be crucial to the development of new therapeutic strategies for CRC.
- Estudo e_COR: mini-relatório da Região CentroPublication . Bourbon, Mafalda; Alves, CatarinaCaracterização da população da Região Centro: Nesta região do país participaram 338 indivíduos, 170 homens e 168 mulheres distribuídos por três classes etárias: 18-34, 35-64 e 65-79 anos. A seleção dos participantes e a sua distribuição geográfica encontra-se descrita na metodologia de amostragem. Os dados foram ponderados por sexo e idade de modo a constituírem uma amostra representativa da região centro, tal como descrito na metodologia de análise.
- Functional analysis of splicing mutations in the IDS gene and the use of antisense oligonucleotides to exploit an alternative therapy for MPS IIPublication . Matos, Liliana; Gonçalves, Vânia; Pinto, Eugénia; Laranjeira, Francisco; Prata, Maria João; Jordan, Peter; Desviat, Lourdes R.; Perez, Belén; Alves, SandraMucopolysaccharidosis II is a lysosomal storage disorder caused by mutations in the IDS gene, including exonic alterations associated with aberrant splicing. In the present work, cell-based splicing assays were performed to study the effects of two splicing mutations in exon 3 of IDS, i.e., c.241C>T and c.257C>T, whose presence activates a cryptic splice site in exon 3 and one in exon 8, i.e., c.1122C>T that despite being a synonymous mutation is responsible for the creation of a new splice site in exon 8 leading to a transcript shorter than usual. Mutant minigene analysis and overexpression assays revealed that SRSF2 and hnRNP E1 might be involved in the use and repression of the constitutive 3' splice site of exon 3 respectively. For the c.1122C>T the use of antisense therapy to correct the splicing defect was explored, but transfection of patient fibroblasts with antisense morpholino oligonucleotides (n=3) and a locked nucleic acid failed to abolish the abnormal transcript; indeed, it resulted in the appearance of yet another aberrant splicing product. Interestingly, the oligonucleotides transfection in control fibroblasts led to the appearance of the aberrant transcript observed in patients' cells after treatment, which shows that the oligonucleotides are masking an important cis-acting element for 5' splice site regulation of exon 8. These results highlight the importance of functional studies for understanding the pathogenic consequences of mis-splicing and highlight the difficulty in developing antisense therapies involving gene regions under complex splicing regulation.
- Effects of physical exercise training in DNA damage and repair activity in humans with different genetic polymorphisms ofhOGG1(Ser326Cys)Publication . Soares, Jorge Pinto; Silva, Ana Inês; Silva, Amélia M.; Almeida, Vanessa; Teixeira, João Paulo; Matos, Manuela; Gaivão, Isabel; Mota, Maria PaulaThe main purpose of this pilot study was to investigate the possible influence of genetic polymorphisms of the hOGG1 (Ser326Cys) gene in DNA damage and repair activity by 8-oxoguanine DNA glycosylase 1 (OGG1 enzyme) in response to 16 weeks of combined physical exercise training. Thirty-two healthy Caucasian men (40-74 years old) were enrolled in this study. All the subjects were submitted to a training of 16 weeks of combined physical exercise. The subjects with Ser/Ser genotype were considered as wild-type group (WTG), and Ser/Cys and Cys/Cys genotype were analysed together as mutant group (MG). We used comet assay in conjunction with formamidopyrimidine DNA glycoslyase (FPG) to analyse both strand breaks and FPG-sensitive sites. DNA repair activity were also analysed with the comet assay technique. Our results showed no differences between DNA damage (both strand breaks and FPG-sensitive sites) and repair activity (OGG1) between genotype groups (in the pre-training condition). Regarding the possible influence of genotype in the response to 16 weeks of physical exercise training, the results revealed a decrease in DNA strand breaks in both groups, a decrease in FPG-sensitive sites and an increase in total antioxidant capacity in the WTG, but no changes were found in MG. No significant changes in DNA repair activity was observed in both genotype groups with physical exercise training. This preliminary study suggests the possibility of different responses in DNA damage to the physical exercise training, considering the hOGG1 Ser326Cys polymorphism.
- Estudo e_COR: mini-relatório da Região de LisboaPublication . Bourbon, Mafalda; Alves, CatarinaCaracterização da população da Região de Lisboa: Nesta região do país participaram 350 indivíduos, 185 homens e 165 mulheres distribuídos por três classes etárias: 18-34, 35-64 e 65-79 anos. A seleção dos participantes e a sua distribuição geográfica encontra-se descrita na metodologia de amostragem. Os dados foram ponderados por sexo e idade de modo a constituírem uma amostra representativa da região de lisboa, tal como descrito na metodologia de análise.
- Data in support of a functional analysis of splicing mutations in the IDS gene and the use of antisense oligonucleotides to exploit an alternative therapy for MPS IIPublication . Matos, Liliana; Gonçalves, Vânia; Pinto, Eugénia; Laranjeira, Francisco; Prata, Maria João; Jordan, Peter; Desviat, Lourdes R.; Pérez, Belén; Alves, SandraThis data article contains insights into the methodology used for the analysis of three exonic mutations altering the splicing of the IDS gene: c.241C>T, c.257C>T and c.1122C>T. We have performed splicing assays for the wild-type and mutant minigenes corresponding to these substitutions. In addition, bioinformatic predictions of splicing regulatory sequence elements as well as RNA interference and overexpression experiments were conducted. The interpretation of these data and further extensive experiments into the analysis of these three mutations and also into the methodology applied to correct one of them can be found in "Functional analysis of splicing mutations in the IDS gene and the use of antisense oligonucleotides to exploit an alternative therapy for MPS II" Matos et al. (2015) [1].
- WHO European Childhood Obesity Surveillance Initiative: health-risk behaviours on nutrition and physical activity in 6-9-year-old schoolchildrenPublication . Wijnhoven, Trudy M.A.; van Raaij, Joop M.A.; Yngve, Agneta; Sjöberg, Agneta; Kunešová, Marie; Duleva, Vesselka; Petrauskiene, Ausra; Rito, Ana I.; Breda, JoãoObjective: To assess to what extent eight behavioural health risks related to breakfast and food consumption and five behavioural health risks related to physical activity, screen time and sleep duration are present among schoolchildren, and to examine whether health-risk behaviours are associated with obesity. Design: Cross-sectional design as part of the WHO European Childhood Obesity Surveillance Initiative (school year 2007/2008). Children’s behavioural data were reported by their parents and children’s weight and height measured by trained fieldworkers. Descriptive statistics and logistic regression analyses were performed. Setting: Primary schools in Bulgaria, Lithuania, Portugal and Sweden; paediatric clinics in the Czech Republic. Subjects: Nationally representative samples of 6–9-year-olds (n 15 643). Results: All thirteen risk behaviours differed statistically significantly across countries. Highest prevalence estimates of risk behaviours were observed in Bulgaria and lowest in Sweden. Not having breakfast daily and spending screen time ≥2 h/d were clearly positively associated with obesity. The same was true for eating ‘foods like pizza, French fries, hamburgers, sausages or meat pies’ >3 d/week and playing outside <1 h/d. Surprisingly, other individual unhealthy eating or less favourable physical activity behaviours showed either no or significant negative associations with obesity. A combination of multiple less favourable physical activity behaviours showed positive associations with obesity, whereas multiple unhealthy eating behaviours combined did not lead to higher odds of obesity. Conclusions: Despite a categorization based on international health recommendations, individual associations of the thirteen health-risk behaviours with obesity were not consistent, whereas presence of multiple physical activity-related risk behaviours was clearly associated with higher odds of obesity.
- Estudo e_COR: mini-relatório da Região do AlentejoPublication . Bourbon, Mafalda; Alves, CatarinaCaracterização da população da Região do Alentejo: Nesta região do país participaram 344 indivíduos, 166 homens e 178 mulheres distribuídos por três classes etárias: 18-34, 35-64 e 65-79 anos. A seleção dos participantes e a sua distribuição geográfica encontra-se descrita na metodologia de amostragem. Os dados foram ponderados por sexo e idade de modo a constituírem uma amostra representativa da região do Alentejo, tal como descrito na metodologia de análise.