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Obstructive Sleep Apnea (OSA) syndrome is characterized by recurrent episodes of apneas and hypopneas during sleep, leading to recurrent intermittent hypoxia and sleep fragmentation. No treated OSA can result in metabolic and cardiovascular diseases. By 2D gel-based proteomics approach we have demonstrated that OSA can cause alterations in the red blood cells (RBC) proteome that may be associated with OSA outcomes. OSA induces alterations in the redox/oligomeric states of RBC proteins such as gyceraldehyde-3-phosphate dehydrogenase (GAPDH) and peroxiredoxin-2 (PRDX2) that can be reverted or modulated by PAP treatment. In this study, we applied a shotgun proteomics strategy to further investigate the RBC proteome from patients with OSA before and after PAP treatment to better understand the regulation of RBC homeostasis in the context of OSA and/or under effect of PAP treatment. As a first approach, RBCs samples, corresponding to Snorers patients as control (n=23) and patients with OSA before and after six months of PAP treatment (n=33/condition) were selected from our biobank1. Samples were randomly pooled (n=3 per group/condition) and lysed 1:6 with 5mM sodium phosphate buffer containing 100 mM of N-ethylmaleimide, a reagent that alkylates free sulfhydryl groups, before haemoglobin depletion by using HemovoidTM system. Depleted samples were alkylated, reduced and digested with trypsin and chymotrypsin. The resulting peptides were cleaned with C18 columns and analysed in triplicate by a Nano High Performance Liquid Chromatography (nanoHPLC) on-line coupled to a high-resolution accurate-mass Orbitrap mass spectrometer (Q Exactive, Thermo Scientific) with a nano electrospray ionization source (nanoESI). The acquired mass spectrometry data were analysed by MaxQuant v1.5.8.3 and Perseus v2.0.3.1 software. The preliminary results corroborated our previous findings by showing that proteins associated with stress response and antioxidant regulatory system were the most changed in OSA RBC compared with Snorers ones. The active catalytic cysteine (Cys 51) in the PRDX2 was identified trioxidized –SO3H almost exclusively in OSA RBC before PAP treatment. Further analyses and validation of these data are in progress, which will certainly provide a better understanding of RBC molecular mechanisms and their proteins/PTMs associated with OSA pathology and/or response to PAP therapy. Project partially supported by the Harvard Medical School-Portugal Program (HMSP-ICJ/0022/2011), the ToxOmics (FCT-UID/BIM/00009/2013) and the Portuguese Mass Spectrometry Network (RNEM). CVC is recipient of FCT doctoral scholarship FCT-SFRH/BD/133511/2017. info:eu-repo/semantics/draft

The 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. N/A

Question: 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

Environmental 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. Work partially funded by Calouste Gulbenkian Foundation and Administração Central do Sistema de Saúde, Portugal. SN, SP worked under FCT fellowships N/A

Obstructive sleep apnea (OSA) is a common sleep-related breathing disorder which is characterized by recurrent occurrence of partial or complete closure of the upper airway during sleep, despite ongoing efforts to breathe. The majority of patients with OSA remain undiagnosed since most of them only come to the attention of a clinician when they complain of daytime sleepiness or when their bed partners report loud snoring or witnessed apnea episodes.Epidemiological studies have indicated that OSA affects 6–13% of the adult population. OSA is multifactorial disease, also considered as metabolic syndrome, which diagnosis in early stages is challenging thus often remain undiagnosed. Recently was found connection between transthyretin (TTR) protein modifications present in human plasma samples and appearance of sleep apnea syndrome1,2 . Mass Spectrometric Immunoassay (MSIA) was successfully applied previously on identification of and quantification of TTR variants present in human serum3. We took advantage on this powerful method to investigate possible modifications of TTR proteoforms in patients with OSAS. N/A

Uncovering mechanisms of unknown pathological mechanisms and body response to applied medication are the drive forces toward personalized medicine. In this post-genomic era, all eyes are tuned to proteomic field, searching for the answers and explanations by investigating the final physiological functional units – proteins and their proteoforms. Development of cutting-edge mass spectrometric technologies and powerful bioinformatics tools, allowed life-science community mining of disease-specific proteins as biomarkers, which are often hidden by high complexity of the samples and/or small abundance. Nowadays, there are several proteomics-based approaches to study the proteome. This chapter focuses on gold standard proteomics strategies and related issues towards candidate biomarker discovery, which may have diagnostic/prognostic as well as mechanistic utility.

Third-generation aromatase inhibitors are more effective than tamoxifen for preventing recurrence in postmenopausal women with hormone-receptor-positive invasive breast cancer. However, it is not known whether anastrozole is more effective than tamoxifen for women with hormone-receptor-positive ductal carcinoma in situ (DCIS). Here, we compare the efficacy of anastrozole with that of tamoxifen in postmenopausal women with hormone-receptor-positive DCIS.In a double-blind, multicentre, randomised placebo-controlled trial, we recruited women who had been diagnosed with locally excised, hormone-receptor-positive DCIS. Eligible women were randomly assigned in a 1:1 ratio by central computer allocation to receive 1 mg oral anastrozole or 20 mg oral tamoxifen every day for 5 years. Randomisation was stratified by major centre or hub and was done in blocks (six, eight, or ten). All trial personnel, participants, and clinicians were masked to treatment allocation and only the trial statistician had access to treatment allocation. The primary endpoint was all recurrence, including recurrent DCIS and new contralateral tumours. All analyses were done on a modified intention-to-treat basis (in all women who were randomised and did not revoke consent for their data to be included) and proportional hazard models were used to compute hazard ratios and corresponding confidence intervals. This trial is registered at the ISRCTN registry, number ISRCTN37546358.Between March 3, 2003, and Feb 8, 2012, we enrolled 2980 postmenopausal women from 236 centres in 14 countries and randomly assigned them to receive anastrozole (1449 analysed) or tamoxifen (1489 analysed). Median follow-up was 7·2 years (IQR 5·6-8·9), and 144 breast cancer recurrences were recorded. We noted no statistically significant difference in overall recurrence (67 recurrences for anastrozole vs 77 for tamoxifen; HR 0·89 [95% CI 0·64-1·23]). The non-inferiority of anastrozole was established (upper 95% CI