Browsing by Author "James, Peter"
Now showing 1 - 10 of 12
Results Per Page
Sort Options
- Facing challenges in Proteomics today and in the coming decade: Report of Roundtable Discussions at the 4th EuPA Scientific Meeting, Portugal, Estoril 2010Publication . Cox, Jurgen; Heeren, Ron M.A.; James, Peter; Jorrin-Novo, Jesús V.; Kolker, Eugene; Levander, Fredrik; Morrice, Nicholas; Picotti, Paola; Righetti, Pier Giorgio; Sánchez, Jean-Charles; Turckl, Christoph W.; Zubarev, Roman; Alexandre, Bruno M.; Corrales, Fernando J; Marko-Varga, György; O'Donovan, Sinead; O'Neil, Serena; Prechl, Jozsef; Tânia, Simões; Weckwerth, Wolfram; Deborah, PenqueRoundtable Discussion sessions were conceived for the first time at the 4th Edition of the EuPA Annual Scientific Meeting that took place in Estoril, Portugal, from the 23rd to 27th of October, 2010. Under the theme “2010: a proteomics odyssey towards the next decades”, young and senior scientists had the opportunity to discuss particular issues of different proteomic areas s distributed by eleven luncheon roundtables moderated by recognized experts in the field. During two hours, the roundtable discussions proceeded in a friendly environment, in which the participants (about 8–10 per roundtable) were encouraged to ask questions and express their opinions without restraint (Fig. 1). The topics covered a range of recurring issues such as the capabilities and limitations of several proteomic methodologies and suggestions on how researchers may better overcome them. The major goal devised by roundtable leaders was to provide independent advice, describe limitations, give practical tips, help set realistic expectations and answer questions from the participants. Participants were also encouraged to be ambitious in generating new solutions or strategies for the future.
- Impacto do fumo do cigarro passivo no proteoma humano: em busca de biomarcadores precoces de risco para a saúdePublication . Neves, Sofia; Pacheco, Solange A.; Vaz, Fátima; Valentim-Coelho, Cristina; Saraiva, Joana; James, Peter; Simões, Tânia; Penque, DeborahOs não-fumadores expostos ao fumo do cigarro passivo ou, simplesmente fumo passivo (FP), apresentam um risco acrescido de desenvolver diversas doenças graves. No entanto, os mecanismos moleculares que explicam estes efeitos continuam pouco esclarecidos, o que reforça a necessidade de identificar biomarcadores capazes de avaliar o risco associado a esta exposição. Neste estudo, analisámos o proteoma do epitélio nasal e do plasma de indivíduos não-fumadores saudáveis expostos ao FP no local de trabalho, num contexto ainda enquadrado pela Lei n.º 37/2007, utilizando uma abordagem proteómica ‘shotgun’ por espectrometria de massa. No epitélio nasal, observámos um aumento de proteínas envolvidas em vias centrais do metabolismo energético, como a Gliceraldeído-3-fosfato desidrogenase (GAPDH) e a Triosefosfato isomerase (TPI1), sugerindo uma possível reprogramação metabólica induzida pela exposição. Identificámos também uma diminuição da tubulina beta-4B (TUBB4B), relacionada com a organização do citoesqueleto, e um aumento da proteína anti-apoptótica SERPINB3, apontando para alterações em processos de morte e sobrevivência celular. No plasma, destacaram-se o aumento da Butirilcolinesterase (BChE) e a diminuição da Proteína de ligação à vitamina D (GC), ambas associadas à resposta a xenobióticos e a processos de lesão tecidular. Foram ainda detetadas alterações em proteínas reguladoras da inflamação sistémica, como C1R, C1QC, HRG e PROS1. A expressão diferencial de APOA4 e SERPINF2 sugere, adicionalmente, a ativação de mecanismos relacionados com risco aterotrombótico. Em conjunto, estes resultados contribuem para aprofundar a compreensão das vias biológicas que ligam a exposição ao fumo passivo ao risco acrescido de cancro e de doenças cardiovasculares, e apresentam um conjunto promissor de potenciais biomarcadores para avaliação do risco associado à exposição ao FP.
- Occupational exposure to environmental tobacco smoke in hospitality venues: are genetic- or proteomics-based biomarkers predictive of respiratory diseases?Publication . Silva, M.J.; Vital, Nádia; Pacheco, Solange; Antunes, Susana; Neves, Sofia; Louro, Henriqueta; Torre, Vukosava M.; Vaz, Fátima; James, Peter; Lopes, Carlos; Marçal, Nelson; Bugalho de Almeida, António; Simões, Tânia; Penque, DeborahEnvironmental tobacco smoke (ETS) is recognized as an occupational hazard in the hospitality industry. Although Portuguese legislation banned smoking in most indoor public spaces, it is still allowed in some restaurants/bars, representing a potential risk to the workers’ health, particularly for chronic respiratory diseases. The aims of this work were to characterize biomarkers of early genetic effects and to disclose proteomic signatures associated to occupational exposure to ETS and with potential to predict respiratory diseases development. A detailed lifestyle survey and clinical evaluation (including spirometry) were performed in 81 workers from Lisbon restaurants. ETS exposure was assessed through the level of PM 2.5 in indoor air and the urinary level of cotinine. The plasma samples were immunodepleted and analysed by 2D-SDSPAGE followed by in-gel digestion and LC-MS/MS. DNA lesions and chromosome damage were analysed innlymphocytes and in exfoliated buccal cells from 19 cigarette smokers, 29 involuntary smokers, and 33 non-smokers not exposed to tobacco smoke. Also, the DNA repair capacity was evaluated using an ex vivo challenge comet assay with an alkylating agent (EMS). All workers were considered healthy and recorded normal lung function. Interestingly, following 2D-DIGE-MS (MALDI-TOF/TOF), 61 plasma proteins were found differentially expressed in ETS-exposed subjects, including 38 involved in metabolism, acute-phase respiratory inflammation, and immune or vascular functions. On the other hand, the involuntary smokers showed neither an increased level of DNA/chromosome damage on lymphocytes nor an increased number of micronuclei in buccal cells, when compared to non-exposed non-smokers. Noteworthy, lymphocytes challenge with EMS resulted in a significantly lower level of DNA breaks in ETS-exposed as compared to non-exposed workers (P<0.0001) suggestive of an adaptive response elicited by the previous exposure to low levels of ETS. Overall, changes in proteome may be promising early biomarkers of exposure to ETS. Likewise, alterations of the DNA repair competence observed upon ETS exposure deserves to be further understood. Work supported by Fundação Calouste Gulbenkian, ACSS and FCT/Polyannual Funding Program.
- Occupational second-hand smoke exposure: A comparative shotgun proteomics study on nasal epithelia from healthy restaurant workersPublication . Neves, Sofia; Pacheco, Solange; Vaz, Fátima; James, Peter; Simões, Tânia; Penque, Deborah; NevesNon-smokers exposed to second-hand smoke (SHS) present risk of developing tobacco smoke-associated pathologies. To investigate the airway molecular response to SHS exposure that could be used in health risk assessment, comparative shotgun proteomics was performed on nasal epithelium from a group of healthy restaurant workers, non-smokers (never and former) exposed and not exposed to SHS in the workplace. HIF1α-glycolytic targets (GAPDH, TPI) and proteins related to xenobiotic metabolism, cell proliferation and differentiation leading to cancer (ADH1C, TUBB4B, EEF2) showed significant modulation in non-smokers exposed. In never smokers exposed, enrichment of glutathione metabolism pathway and EEF2-regulating protein synthesis in genotoxic response were increased, while in former smokers exposed, proteins (LYZ, ATP1A1, SERPINB3) associated with tissue damage/regeneration, apoptosis inhibition and inflammation that may lead to asthma, COPD or cancer, were upregulated. The identified proteins are potential response and susceptibility/risk biomarkers for SHS exposure.
- Occupational secondhand smoke effect on nasal epithelial proteomaPublication . Pacheco, Solange; Vaz, Fatima; Torres, Vukosava; Penque, Deborah; Simões, Tania; James, PeterThe tobacco is one of the biggest public health threats, smoking kills more than 7 million people/year worldwide and more than 890,000 are deaths resulting from exposure to Secondhand Smoke (SHS). In adults, SHS is associated to cardiovascular and respiratory diseases, including coronary heart disease and lung cancer, through pathological and molecular mechanisms not yet understood. In this study, we aimed to investigate the SHS effects on nasal epithelial proteome in exposed workers.
- Occupational secondhand smoke exposure - A proteomic analysisPublication . Neves, Sofia; Pacheco, Solange; Vaz, Fátima; Simões, Tania; James, Peter; Simões, Tânia; Penque, DeborahBackground: WHO have stated that near 900 000 deaths per year result from exposure to Second- Hand Smoke (SHS). SHS exposure has been linked to cancer, respiratory and cardiovascular diseases and diabetes. However, the associated underlying molecular mechanisms remain to be elucidated. The objective of this proteomics study is to uncover putative key molecules involved in these mechanisms that can be used to predict and monitor diseases risks associated with occupational SHS exposure. Methods: In total, 25 Lisbon restaurants agreed to participate. Nasal epithelium and urine samples were collected from their employees (n=52) for proteomics analysis and cotinine evaluation of SHS exposure, respectively. The subjects were classified as never smoker (N), former smoker (F) and smoker (S); exposed (NE=11; FE=10; SE=4) or non-exposed (N=11; F=8; S=8) to SHS. All subjects were healthy and showed no significant differences in parameters like age, time in the workplace, tobacco smoking habits and spirometry evaluation of pulmonary function. Urine cotinine levels showed significantly elevated in the exposed subjects compared to non-exposed, confirming SHS exposure. Nasal epithelium samples were analyzed by shotgun proteomics using an ESI-LTQOrbitrap mass spectrometer. The “MS raw data” was submitted to “PatternLab for Proteomics” software, with “Comet” search machine algorithm, from where the identified proteins were submitted to a “ClueGO” functional annotation & enrichment analyses in “Cytoscape” software, with the propose to shed some light about the molecular biology involved in the cellular response to the SHS exposition. Results: In NE subjects the SHS is associated with the biologic terms of “Lactate dehydrogenase complex” and “Pentose-Phosphatase Shunt”, also with “Glutathione peroxidase activity” and “Tcell apoptotic process”. At the other end the FE individuals present a specific proteome enriched in biologic information with terms as the “L-Lactate dehydrogenase complex” and the “Peroxisome” as was expected by the results above for the NE cohort; but there were also other different terms as: “Peripheral T cell lymphoma”, “Central carbon metabolism in cancer”, “Myelodysplastic syndrome”, “Monocyte & Granulocyte & Macrophage & Leukocyte Chemotaxis”, Nucleossome, variant H3.1-H2A2-H2B.1&Others” and finally “DNA replication-dependent chromatin assembly”. Conclusions: Proteome of nasal epithelium seems to be modulated by SHS exposure and this is a different and perhaps cumulative process between NE and FE individuals.
- Occupational secondhand smoke exposure may modify the proteoma expression of human nasal epitheliumPublication . Neves, Sofia; Pacheco, Solange; Vaz, Fátima; Martins, Inês; James, Peter; Simões, Tânia; Penque, DeborahThe tobacco is one of the biggest public health threats, smoking kills more than 7 million people/year worldwide and more than 890,000 are deaths resulting from exposure to Second Hand Smoke (SHS). In adults, SHS is associated to cardiovascular and respiratory diseases, including coronary heart disease and lung cancer, through pathological and molecular mechanisms not yet understood. In this study, we aimed to investigate the SHS effects on airway proteome in exposed workers. Nasal epithelium was collected from hospitality workers (non-smokers=40; smokers=12) long-term exposed and non-exposed to SHS at the workplace. Samples were analyzed by shotgun proteomics using an ESI-LQT Orbitrap XL mass spectrometer. The generated MS raw data was submitted to ‘PatternLab for Proteomics 4.0’ for peptide identification and relative quantification by label-free - extracted ion chromatograms (XIC). Golden rules were applied to obtain reliable data such as the inferred proteins must have at least one unique peptide identified to be considered and be detected in at least 80% of the cohort. Two proteins were found to be differentially expressed in the no-smokers exposed to SHS compared with the control: BPI fold-containing family A member 1 (BPIFA1) and Heat shock Protein Beta-1 (HSPB1). The first protein plays a role in the airway inflammatory response after exposure to irritants substances and the second is associated as a regulator of actin filament dynamics. Our findings support the indication that in non-smokers the prolonged exposure to SHS can lead to airway proteome modulation. When validated, the uncovered proteins can be promising candidates to “susceptibility/risk” and/or “response” biomarkers for SHS exposure.
- Occupational secondhand smoke exposure may modify the proteoma expression of human nasal epitheliumPublication . Neves, Sofia; Pacheco, Solange; Vaz, Fátima; Torres, Vukosava Milic; James, Peter; Simões, Tânia; Penque, DeborahThe tobacco is one of the biggest public health threats, smoking kills more than 7 million people/year worldwide and more than 890,000 are deaths resulting from exposure to Second Hand Smoke (SHS). In adults, SHS is associated to cardiovascular and respiratory diseases, including coronary heart disease and lung cancer, through pathological and molecular mechanisms not yet understood. We aimed to investigate the SHS effects on airway proteome in exposed workers. Nasal epithelium was collected from hospitality workers (non-smokers=40; smokers=12), long-term exposed and non-exposed to SHS at the workplace. Samples were analyzed by shotgun proteomics using an ESI-LQT Orbitrap XL mass spectrometer. The generated MS raw data was submitted to ‘PatternLab for Proteomics’ for peptide identification and relative quantification by label-free - extracted ion chromatograms (XIC). Golden rules were applied to obtain reliable data: in the identification of a protein at least one unique peptide must had to be present in more than 80% of the individuals, and consequently each inferred protein had to be detected in 80% to 100% of the cohort. Two proteins were found to be differentially expressed in the no-smokers exposed to SHS compared with the control: BPI fold-containing family A member 1 (BPIFA1) and Heat shock Protein Beta-1 (HSPB1). The first protein plays a role in the airway inflammatory response after exposure to irritants substances and the second is associated as a regulator of actin filament dynamics. Our findings support the indication that in non-smokers the prolonged exposure to SHS can lead to airway proteome modulation. When validated, the uncovered proteins can be promising candidates to “susceptibility/risk” and/or “predictive” biomarkers for SHS exposure.
- Occupational smoke exposure may modify the expression of respiratory tract proteomaPublication . Neves, Sofia; Pacheco, Solange; Vaz, Fatima; Torres, Vukosava Milic; James, Peter; Simões, Tânia; Penque, DeborahQuestion: Tobacco is one of the biggest public health threats. Smoking kills more than 7 million people/year worldwide and more than 890,000 of deaths resulted from exposure to Second Hand Smoke (SHS). SHS is associated to cardiovascular and respiratory diseases, including coronary heart disease and lung cancer, through pathological and molecular mechanisms not yet understood. In Portugal, the partial tobacco smoking ban legislation in public venues allows smoking in restaurants with designed rooms larger than 100m2. We aimed to investigate the SHS effects on the respiratory tract proteome from exposed workers. Methods: Nasal epithelia was collected from hospitality workers (non-smokers=40; smokers=12), long-term exposed and non-exposed to SHS at the workplace. All subjects were healthy with normal spirometry values (FVE1/FVC <70%) for pulmonary diseases. Non-smokers exposed presented higher levels of urinary cotinine after working, confirming SHS exposure. Samples were analyzed by shotgun proteomics using ESI-LQT Orbitrap XL MS and the generated MS raw data by the PatternLab for Proteomics. Proteins were investigated by DAVIDv6.8 and Reactome Pathway databases. Results: Two proteins were found to be differentially expressed in the nonsmokers exposed to SHS compared with control: Heat shock Protein Beta-1 (HSPB1) and BPI fold-containing family A member 1 (BPIFA1). HSPB1 plays a role in the stress resistance and actin organization and BPIFA1 is involved in the airway inflammatory response after exposure to irritants by attracting macrophages and neutrophils, and it is also associated with tumor progression. Conclusions: In health non-smokers, prolonged exposure to SHS can lead to respiratory tract proteome modulation associated with airway inflammatory/stress response to toxic substances. When validated, the uncovered proteins can be promising candidate biomarkers of exposure/effect for clinical assessment of workers occupationally exposed to SHS.
- Second-hand smoke exposure effects on nasal epithelia proteomePublication . Neves, Sofia; Pacheco, Solange; Vaz, Fatima; James, Peter; Penque, DeborahEnvironmental second-hand smoke exposure (SHS) results in a statistically significant increase in the risk of diseases such cardiovascular diseases and lung cancer. Cigarette smoke contains thousands of constituents, including several carcinogens and cytotoxic chemicals that orchestrate chronic inflammatory responses and destructive remodeling events1,2. In this work, our main objective is to uncover biomarkers of SHS exposure effects by investigating the proteome of nasal epithelia from health subjects occupationally long–term exposed to SHS.