Identificação de potenciais biomarcadores de doença de Alzheimer no plasma usando FTIR

Mestrado em Biomedicina MolecularCurrent clinical AD diagnosis criteria used to identify the disorder in patients who have already overt the advanced stage of dementia. This is too late for some kind of successful disease adjustment and consequently, early recognition of AD needs to be improved. Therefore, it is really needed different approaches for discovering new AD biomarkers, such as the application of metabolomics techniques (e.g. FTIR). The potential of FTIR in the clinical field has recently received particular attention, since it uses vibration frequencies of molecules present in the analysed sample to produce a metabolic fingerprint, which is then specific for each sample. This dissertation aims to identify biochemical alterations that might be related to dementia, being possible to distinguish between control and disease groups of plasma samples, through application of FTIR methodology at the 4000-600 cm-1 spectral region. Using plasma samples makes the process being minimally invasive, with other relevant clinical advantages. Besides the collection of blood samples used in this project, the volunteers were also submitted to cognitive evaluation trough Mini Mental State Examination (MMSE) and Clinical Dementia Rating (CDR), with other relevant clinical information being also collected. All the analysed samples were matched by age and sex. For a better discrimination between control and disease samples, Principal Component Analysis (PCA) was applied to spectra data at specific regions, namely 3500-2700 cm-1, 1700-1400 cm-1, and 1200-900 cm-1. This allowed to identify the main pathological changes that occurred at the biochemical level during neurodegenerative disorder development. In the former spectral region, disease samples presented a higher content of saturated lipids in relation to the unsaturated ones, which translates in a high potential brain damage. Besides, it was also noted the presence of carboxylic acids that are usually related to lipid hyperoxidation, production of reactive carbonyls and proteins structural and functional alterations. In turn, the spectral region 1700-1400 cm-1 allowed to identify differences in protein conformation between control and disease samples, and these last ones were still related with occurrence of protein aggregates. In other hand, the 1200-900 cm-1 region could be associated to cellular damage provoked by oxidative stress in disease samples. As an important note to take, FTIR analysis could have the potential to be applied in future not only for cognitive impairment diagnosis but also for identification of disease stage and prognostic evaluation, besides assessment of disease developing risk for control subjects.Os atuais critérios clínicos de diagnóstico da doença de Alzheimer geralmente identificam a patologia apenas quando os pacientes já atingiram a fase de demência, ou seja, o estágio mais avançado da doença. Isto revela-se demasiado tarde para que qualquer tipo de terapêutica seja bem-sucedida e, consequentemente é necessário desenvolver uma metodologia de diagnóstico que permita um reconhecimento mais precoce da doença. Desta forma, é preciso adotar diferentes abordagens para a descoberta de novos biomarcadores para a doença de Alzheimer, tais como a utilização de técnicas de metabolómica (ex. FTIR). O potencial do FTIR no campo clínico recebeu recentemente particular atenção, sendo que esta técnica utiliza as frequências de vibração das moléculas presentes na amostra analisada para produzir uma “impressão digital” metabólica, a qual é específica para cada amostra. Esta dissertação tem como objetivo a identificação de potenciais alterações bioquímicas que possam estar relacionadas com demência, tornando possível a distinção entre grupos controlo e de doentes no seio do conjunto de amostras de plasma analisadas. A utilização de amostras de plasma torna o processo minimamente invasivo, de entre outras vantagens clínicas. Para além da colheita das amostras de sangue utilizadas neste projeto, os voluntários foram submetidos a uma avaliação cognitiva através da realização do MMSE e do CDR, com outra informação clínica relevante a ser também recolhida. Todas as amostras analisadas foram emparelhadas de acordo com a idade e o sexo. Para uma melhor distinção entre amostras controlo e de doentes foi aplicada a metodologia de PCA aos dados dos espectros obtidos em regiões específicas, nomeadamente em 3500-2700 cm-1, 1700-1400 cm-1, e 1200-900 cm-1. Isto permitiu identificar as principais alterações patológicas que ocorrem a nível bioquímico durante o desenvolvimento da doença neurodegenerativa. Na primeira região espectral referida, as amostras dos doentes apresentaram um maior conteúdo de lípidos saturados comparativamente aos não saturados, o que se traduz num potencial risco cerebral maior. Para além disso, foi observada a presença de ácidos carboxílicos, usualmente relacionados com hiperoxidação de lípidos, produção de carbonilos reativos e alterações estruturais e funcionais de proteínas. Por sua vez, a região espectral 1700-1400 cm-1 permitiu identificar diferenças na conformação de proteínas entre amostras controlo e de doentes, tendo estas últimas sido ainda relacionadas com a ocorrência de agregados proteicos. Por outro lado, a região 1200-900 cm-1 pôde ser relacionada com presença de danos celulares provocados por stress oxidativo nas amostras de doentes. Como importante nota a reter, a análise por FTIR pode ter o potencial para ser aplicada no futuro não apenas para diagnóstico de disfunção cognitiva, mas também para identificação do estádio da doença e realização de prognósticos, para além da avaliação do risco de desenvolvimento da doença em sujeitos controlo

Mechanical pruning and soil organic amending in two terroirs. Effects on wine chemical composition and sensory profile

The knowledge about the interaction between mechanical pruning and soil organic amending is still scarce. This study aimed to examine the effects of the interaction between these two practices on wine quality. Syrah grapes from two trial fields in Portugal subjected to two different pruning systems (mechanical pruning; hand spur pruning) and five different organic amendment treatments (control, biochar, municipal solid waste compost, cattle manure, and sewage sludge) were harvested and vinified for four years. Mechanical pruning significantly reduced wine alcoholic strength, pH, and total anthocyanins. Mechanical pruning and organic amendments, tendentially reduced wine total phenols and tannin power, known as an “estimation of the astringency potential of the wines”. Tasters found low but significant differences in global appreciation with the pruning system. Sludge tended to reduce wine global appreciation more than municipal solid waste compost and cattle manure, while biochar had no effect on tasters’ preference when compared to the control. There was strong relation between yield and tasters’ preference only above 6 kg/vine and 8 kg/vine depending on the terroir. Mechanical pruning tendentially has significant effects on wine quality when yield raises above a certain level. Thus, with this pruning system, the choice of the organic amendment and its amount must be done considering the destiny of the produced grapes. To the best of our knowledge, effects of the interaction of mechanical pruning with soil organic amending on wine quality are a noveltyinfo:eu-repo/semantics/publishedVersio

Portugal as a Case Study

Introduction: The International Health Regulations (IHR) were developed to prepare countries to deal with public health emergencies. The spread of SARS-CoV-2 underlined the need for international coordination, although few attempts were made to evaluate the integrated implementation of the IHR's core capacities in response to the COVID-19 pandemic. The aim of this study was to evaluate whether IHR shortcomings stem from non-compliance or regulatory issues, using Portugal as a European case study due to its size, organization, and previous discrepancies between self-reporting and peer assessment of the IHR's core capacities. Methods: Fifteen public health medical residents involved in contact tracing in mainland Portugal interpreted the effectiveness of the IHR's core capabilities by reviewing the publicly available evidence and reflecting on their own field experience, then grading each core capability according to the IHR Monitoring Framework. The assessment of IHR enforcement considered efforts made before and after the onset of the pandemic, covering the period up to July 2021. Results: Four out of nine core IHR capacities (surveillance; response; risk communication; and human resource capacity) were classified as level 1, the lowest. Only two were graded level 3 (preparedness; and laboratory), the highest. The remaining three) (national legislation, policy & financing; coordination and national focal point communication; and points of entry) were classified as level 2. Conclusion: Portugal exemplifies the extent to which implementation of the IHR was not fully achieved, which has resulted in the underperformance of several core capacities. There is a need to improve preparedness and international cooperation in order to harmonize and strengthen the global response to public health emergencies, with better political, institutional, and financial support.publishersversionpublishersversionpublishe

Measuring adherence to inhaled control medication in patients with asthma: Comparison among an asthma app, patient self‐report and physician assessment

Background Previous studies have demonstrated the feasibility of using an asthma app to support medication management and adherence but failed to compare with other measures currently used in clinical practice. However, in a clinical setting, any additional adherence measurement must be evaluated in the context of both the patient and physician perspectives so that it can also help improve the process of shared decision making. Thus, we aimed to compare different measures of adherence to asthma control inhalers in clinical practice, namely through an app, patient self-report and physician assessment. Methods This study is a secondary analysis of three prospective multicentre observational studies with patients (≥13 years old) with persistent asthma recruited from 61 primary and secondary care centres in Portugal. Patients were invited to use the InspirerMundi app and register their inhaled medication. Adherence was measured by the app as the number of doses taken divided by the number of doses scheduled each day and two time points were considered for analysis: 1-week and 1-month. At baseline, patients and physicians independently assessed adherence to asthma control inhalers during the previous week using a Visual Analogue Scale (VAS 0–100). Results A total of 193 patients (72% female; median [P25–P75] age 28 [19–41] years old) were included in the analysis. Adherence measured by the app was lower (1 week: 31 [0–71]%; 1 month: 18 [0–48]%) than patient self-report (80 [60–95]) and physician assessment (82 [51–94]) (p 0.05). There was a moderate correlation between patient self-report and physician assessment (ρ = 0.596, p < 0.001). Conclusions Adherence measured by the app was lower than that reported by the patient or the physician. This was expected as objective measurements are commonly lower than subjective evaluations, which tend to overestimate adherence. Nevertheless, the low adherence measured by the app may also be influenced by the use of the app itself and this needs to be considered in future studies.info:eu-repo/semantics/publishedVersio

Determinants of HIV late presentation among men who have sex with men in Portugal (2014–2019): who’s being left behind?

Introduction: HIV late presentation (LP) remains excessive in Europe. We aimed to analyze the factors associated with late presentation in the MSM population newly diagnosed with HIV in Portugal between 2014 and 2019. Methods: We included 391 newly HIV-1 diagnosed Men who have Sex with Men (MSM), from the BESTHOPE project, in 17 countrywide Portuguese hospitals. The data included clinical and socio-behavioral questionnaires and the viral genomic sequence obtained in the drug resistance test before starting antiretrovirals (ARVs). HIV-1 subtypes and epidemiological surveillance mutations were determined using different bioinformatics tools. Logistic regression was used to estimate the association between predictor variables and late presentation (LP). Results: The median age was 31 years, 51% had a current income between 501-1,000 euros, 28% were migrants. 21% had never been tested for HIV before diagnosis, with 42.3% of MSM presenting LP. 60% were infected with subtype B strains. In the multivariate regression, increased age at diagnosis, higher income, lower frequency of screening, STI ever diagnosed and higher viral load were associated with LP. Conclusion: Our study suggests that specific subgroups of the MSM population, such older MSM, with higher income and lower HIV testing frequency, are not being targeted by community and clinical screening services. Overall, targeted public health measures should be strengthened toward these subgroups, through strengthened primary care testing, expanded access to PrEP, information and promotion of HIV self-testing and more inclusive and accessible health services.info:eu-repo/semantics/publishedVersio

HIV-1-Transmitted Drug Resistance and Transmission Clusters in Newly Diagnosed Patients in Portugal Between 2014 and 2019

Objective: To describe and analyze transmitted drug resistance (TDR) between 2014 and 2019 in newly infected patients with HIV-1 in Portugal and to characterize its transmission networks. Methods: Clinical, socioepidemiological, and risk behavior data were collected from 820 newly diagnosed patients in Portugal between September 2014 and December 2019. The sequences obtained from drug resistance testing were used for subtyping, TDR determination, and transmission cluster (TC) analyses. Results: In Portugal, the overall prevalence of TDR between 2014 and 2019 was 11.0%. TDR presented a decreasing trend from 16.7% in 2014 to 9.2% in 2016 (p for-trend = 0.114). Multivariate analysis indicated that TDR was significantly associated with transmission route (MSM presented a lower probability of presenting TDR when compared to heterosexual contact) and with subtype (subtype C presented significantly more TDR when compared to subtype B). TC analysis corroborated that the heterosexual risk group presented a higher proportion of TDR in TCs when compared to MSMs. Among subtype A1, TDR reached 16.6% in heterosexuals, followed by 14.2% in patients infected with subtype B and 9.4% in patients infected with subtype G. Conclusion: Our molecular epidemiology approach indicates that the HIV-1 epidemic in Portugal is changing among risk group populations, with heterosexuals showing increasing levels of HIV-1 transmission and TDR. Prevention measures for this subpopulation should be reinforced.info:eu-repo/semantics/publishedVersio

Cabbage and fermented vegetables : From death rate heterogeneity in countries to candidates for mitigation strategies of severe COVID-19

Large differences in COVID-19 death rates exist between countries and between regions of the same country. Some very low death rate countries such as Eastern Asia, Central Europe, or the Balkans have a common feature of eating large quantities of fermented foods. Although biases exist when examining ecological studies, fermented vegetables or cabbage have been associated with low death rates in European countries. SARS-CoV-2 binds to its receptor, the angiotensin-converting enzyme 2 (ACE2). As a result of SARS-CoV-2 binding, ACE2 downregulation enhances the angiotensin II receptor type 1 (AT(1)R) axis associated with oxidative stress. This leads to insulin resistance as well as lung and endothelial damage, two severe outcomes of COVID-19. The nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is the most potent antioxidant in humans and can block in particular the AT(1)R axis. Cabbage contains precursors of sulforaphane, the most active natural activator of Nrf2. Fermented vegetables contain many lactobacilli, which are also potent Nrf2 activators. Three examples are: kimchi in Korea, westernized foods, and the slum paradox. It is proposed that fermented cabbage is a proof-of-concept of dietary manipulations that may enhance Nrf2-associated antioxidant effects, helpful in mitigating COVID-19 severity.Peer reviewe

Nrf2-interacting nutrients and COVID-19 : time for research to develop adaptation strategies

There are large between- and within-country variations in COVID-19 death rates. Some very low death rate settings such as Eastern Asia, Central Europe, the Balkans and Africa have a common feature of eating large quantities of fermented foods whose intake is associated with the activation of the Nrf2 (Nuclear factor (erythroid-derived 2)-like 2) anti-oxidant transcription factor. There are many Nrf2-interacting nutrients (berberine, curcumin, epigallocatechin gallate, genistein, quercetin, resveratrol, sulforaphane) that all act similarly to reduce insulin resistance, endothelial damage, lung injury and cytokine storm. They also act on the same mechanisms (mTOR: Mammalian target of rapamycin, PPAR gamma:Peroxisome proliferator-activated receptor, NF kappa B: Nuclear factor kappa B, ERK: Extracellular signal-regulated kinases and eIF2 alpha:Elongation initiation factor 2 alpha). They may as a result be important in mitigating the severity of COVID-19, acting through the endoplasmic reticulum stress or ACE-Angiotensin-II-AT(1)R axis (AT(1)R) pathway. Many Nrf2-interacting nutrients are also interacting with TRPA1 and/or TRPV1. Interestingly, geographical areas with very low COVID-19 mortality are those with the lowest prevalence of obesity (Sub-Saharan Africa and Asia). It is tempting to propose that Nrf2-interacting foods and nutrients can re-balance insulin resistance and have a significant effect on COVID-19 severity. It is therefore possible that the intake of these foods may restore an optimal natural balance for the Nrf2 pathway and may be of interest in the mitigation of COVID-19 severity

Worldwide trends in underweight and obesity from 1990 to 2022: a pooled analysis of 3663 population-representative studies with 222 million children, adolescents, and adults

Background Underweight and obesity are associated with adverse health outcomes throughout the life course. We estimated the individual and combined prevalence of underweight or thinness and obesity, and their changes, from 1990 to 2022 for adults and school-aged children and adolescents in 200 countries and territories. Methods We used data from 3663 population-based studies with 222 million participants that measured height and weight in representative samples of the general population. We used a Bayesian hierarchical model to estimate trends in the prevalence of different BMI categories, separately for adults (age ≥20 years) and school-aged children and adolescents (age 5–19 years), from 1990 to 2022 for 200 countries and territories. For adults, we report the individual and combined prevalence of underweight (BMI &lt;18·5 kg/m2) and obesity (BMI ≥30 kg/m2). For school&#x2;aged children and adolescents, we report thinness (BMI &lt;2 SD below the median of the WHO growth reference) and obesity (BMI &gt;2 SD above the median). Findings From 1990 to 2022, the combined prevalence of underweight and obesity in adults decreased in 11 countries (6%) for women and 17 (9%) for men with a posterior probability of at least 0·80 that the observed changes were true decreases. The combined prevalence increased in 162 countries (81%) for women and 140 countries (70%) for men with a posterior probability of at least 0·80. In 2022, the combined prevalence of underweight and obesity was highest in island nations in the Caribbean and Polynesia and Micronesia, and countries in the Middle East and north Africa. Obesity prevalence was higher than underweight with posterior probability of at least 0·80 in 177 countries (89%) for women and 145 (73%) for men in 2022, whereas the converse was true in 16 countries (8%) for women, and 39 (20%) for men. From 1990 to 2022, the combined prevalence of thinness and obesity decreased among girls in five countries (3%) and among boys in 15 countries (8%) with a posterior probability of at least 0·80, and increased among girls in 140 countries (70%) and boys in 137 countries (69%) with a posterior probability of at least 0·80. The countries with highest combined prevalence of thinness and obesity in school-aged children and adolescents in 2022 were in Polynesia and Micronesia and the Caribbean for both sexes, and Chile and Qatar for boys. Combined prevalence was also high in some countries in south Asia, such as India and Pakistan, where thinness remained prevalent despite having declined. In 2022, obesity in school-aged children and adolescents was more prevalent than thinness with a posterior probability of at least 0·80 among girls in 133 countries (67%) and boys in 125 countries (63%), whereas the converse was true in 35 countries (18%) and 42 countries (21%), respectively. In almost all countries for both adults and school-aged children and adolescents, the increases in double burden were driven by increases in obesity, and decreases in double burden by declining underweight or thinness. Interpretation The combined burden of underweight and obesity has increased in most countries, driven by an increase in obesity, while underweight and thinness remain prevalent in south Asia and parts of Africa. A healthy nutrition transition that enhances access to nutritious foods is needed to address the remaining burden of underweight while curbing and reversing the increase in obesit

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements