Role of apoptotic regulators in the protective effect of acetic acid against ethanol - induced cell death in Saccharomyces cerevisiae

Dissertação de mestrado em Bioquímica AplicadaEthanol is a well-know end product of alcoholic fermentation carried by Saccharomyces cerevisiae. At relatively low concentrations it is an inhibitor of yeast growth, while, high ethanol concentrations reduces cell viability (Birch and Walker, 2000). During fermentation, acetic acid produced by yeast metabolism may accumulate in growth medium and enhance ethanol toxicity (Gibson et al., 2007). By contrast, studies conducted in our laboratory have shown that S. cerevisiae cells exposed simultaneously to toxic concentrations of ethanol and low concentrations of acetic acid displayed a higher survival than cells treated only with ethanol (Vieira et al., unpublished results). Later, Trindade showed that Hog1p and Fps1p, involved in acetic acid resistance, do not have a role in the protective effect of acetic acid seen by Vieira (Trindade, 2009). Subsequently, it was also shown that the osmolyte trehalose, the heat shock protein Hsp12p, the MAP kinase Slt2/MPK1 and functional mitochondria play a role in the protection by the undissociated form of acetic acid against ethanol induced cytotoxicity (Afonso, 2011). With the present master thesis we intended to further understand how acetic acid protects against ethanol-induced cytotoxicity, determining the role of known apoptotic regulators in this protection. We also intended to characterize the death of cells exposed to toxic concentrations of ethanol since this knowledge is important to dissect the role of acetic acid in its inhibition. This study showed that the deletion in CYC3 or ATP2, ATP10 and NUC1 abolish and decrease, respectively, the protection of acetic acid against loss of cell viability, loss of plasma membrane integrity and the accumulation of superoxide anion, induced by ethanol. Death induced by ethanol was associated with typical markers of apoptosis but also of necrosis. In particular caspase activation and exposure of phosphatidylserine was detected mainly in cells with compromised plasma membrane, but also HMGB1/Nhp6A translocates from the nucleus to the cytosol in response to ethanol. Moreover, the absence of known apoptotic regulatory proteins abrogate (atp2Δ, atp10Δ, cyc3Δ, por1Δ, cpr3Δ and yca1Δ mutants), exacerbate (aif1Δ, nuc1Δ, and cyc1Δcyc7Δ mutants) or have no effect (pep4Δ mutant) in ethanol induced cell death. Altogether the results obtained with the mutants phenotypes and the characterization of cell death markers indicate that ethanol triggers a regulated cell death process that shares features typical of both apoptosis and necrosis. Whether the necrotic features reflect a programmed necrosis or a secondary necrosis of apoptotic cells committed to death in response to ethanol, requires further studies.O etanol é um produto final bem conhecido da fermentação alcoólica conduzida por Saccharomyces cerevisiae . Em concentrações relativamente baixas é um inibidor do crescimento da levedura, enquanto em concentrações elevadas induz perda da viabilidade celular (Birch e Walker, 2000). Durante a fermentação, o ácido acético produzido pelo metabolismo da levedura pode acumular-se no meio extracelular e aumentar a toxicidade de etanol (Gibson et al., 2007). Por outro lado, estudos realizados no nosso laboratório mostraram que células de S. cerevisiae simultaneamente expostas a concentrações tóxicas de etanol e baixas concentrações de ácido acético exibiam uma maior sobrevivência do que aquelas tratadas apenas com etanol. (Vieira et. al., resultados não publicados). Mais tarde, Trindade mostrou que as proteínas Hog1p e Fps1p, envolvidas na resistência ao ácido acético, não apresentam um papel no efeito protetor do ácido acético observado por Vieira (Trindade, 2009). Posteriormente, mostrou-se que o osmólito trealose, a proteína de choque térmico Hsp12p, a MAP quinase Slt2/MPK1 e mitocôndrias funcionais têm um papel na proteção da forma não dissociada do ácido acético contra citotoxicidade induzida por etanol (Afonso, 2011). Com a presente dissertação de mestrado pretendeu-se compreender melhor como o ácido acético protege contra a citotoxicidade induzida por etanol, determinando o papel de reguladores conhecidos da apoptose nesta proteção. Também se pretendeu caracterizar a morte de células expostas a concentrações tóxicas de etanol uma vez que este conhecimento é importante para dissecar o papel do ácido acético, na sua inibição. Este estudo mostrou que a ausência dos genes CYC3 ou ATP2, ATP10 e NUC1 elimina e diminui, respetivamente, a proteção do ácido acético contra a perda induzida pelo etanol da viabilidade celular, da integridade da membrana plasmática e da acumulação de anião superóxido. A morte induzida pelo etanol foi acompanhada de marcadores celulares típicos de apoptose, mas também de necrose. Em particular foi detetada ativação de caspases e exposição de fosfatidilserina principalmente em células com a membrana plasmática comprometida, mas também a translocação do HMGB1/Nhp6A do núcleo para o citosol em resposta ao etanol. Adicionalmente, a ausência de reguladores apoptóticos reduziram (nos mutantes atp2Δ, atp10Δ, cyc3Δ, por1Δ, cpr3Δ e yca1Δ), potenciaram (nos mutantes aif1Δ, nuc1Δ, e cyc1Δcyc7Δ ) ou não afetaram a sobrevivência celular (no mutante pep4Δ) em resposta ao etanol. Os resultados respeitantes aos fenótipos dos mutantes estudados e à caracterização dos marcadores de morte celular indicam que o etanol induz uma morte regulada que partilha características típicas de apoptose e necrose. Se as características de necrose refletem uma necrose programada ou uma necrose secundária de células que desencadearam um processo de morte apoptótico, requer mais estudos no futuro.Fundação para a Ciência e a Tecnologia (FCT) - Projeto PEst- C/BIA/UI4050/2011, FCOMP-01-0124-FEDER-007047 e PTDC/AGR- ALI/102608/2008.FEDER através do POFC–COMPET

eHealth promoting stoma self-care for people with an elimination ostomy: focus group study

Background: The construction of an elimination stoma has a physical, psychological, and social impact on the person. The development of stoma self-care competence contributes to the adaptation to a new health condition and improvement of quality of life. eHealth refers to everything associated with information and communication technology and health care, including telemedicine, mobile health, and health informatics. The use of eHealth platforms by the person with an ostomy, as a digital application that includes websites and mobile phone apps, can bring scientific knowledge and well-informed practices to individuals, families, and communities. It also allows functionalities that enable the person to describe and identify early signs and symptoms and precursors of complications and to be guided to an adequate health response for their problems. Objective: This study aimed to define the most relevant content and features to promote ostomy self-care integrated into an eHealth platform as a digital app or website to be used by patients for self-management of stoma care. Methods: We developed a descriptive, exploratory study with a qualitative approach using the focus group methodology, which was oriented to reach a consensus of at least 80%. A convenience sample of 7 participants consisting of stomatherapy nurses was used. The focus group discussion was recorded, and field notes were taken. The focus group meeting was fully transcribed, and a qualitative analysis was performed. The research question was: Which content and features for ostomy self-care promotion should be integrated into an eHealth platform as a digital app or website? Results: An eHealth platform, which can be a smartphone app or website, for people with ostomy should provide content aimed at promoting self-care, namely in the field of knowledge and self-monitoring, as well as the possibility of interacting with a stomatherapy care nurse. Conclusions: The stomatherapy nurse has a decisive role in promoting adaptation to life with a stoma, namely through the promotion of stoma self-care. Technological evolution has emerged as a useful tool to enhance nursing interventions and promote self-care competence. The development of an eHealth platform aimed at promoting ostomy self-care should include the capabilities for telehealth and help with decision-making regarding self-monitoring and seeking differentiated care.info:eu-repo/semantics/publishedVersio

Eu conto! – Experiências e perceções sobre o trabalho e o desemprego na primeira pessoa

[Excerto] No presente texto visa-se explicitar as reflexões desenvolvidas no quadro do grupo de trabalho “Desemprego e desigualdades sociais: desafios para a governação integrada” (DesGov) do Forum Govint. O mercado de trabalho tem vindo a ser objeto de transformações significativas na Europa, em geral, e em Portugal, em particular. Em associação com a mais recente crise económica nos países Ocidentais, que remonta a finais de 2007, manifesta tendências crescentes de rutura e de coexistência de situações heterogéneas para as quais importa estar atento. Estamos perante mercados de trabalho constituídos por múltiplos segmentos que se diferenciam entre si em função de fatores como o nível de escolaridade e de qualificação, o local de residência, a idade, o sexo, a origem social, etc. e que, porque distintos, exigem respostas diferenciadas ao nível do seu enquadramento laboral, quer da parte do Estado, quer do terceiro setor, quer ainda do setor lucrativo. [...

Nurse’s contribution to breastfeeding in basic attention / Contribuição do enfermeiro ao aleitamento materno na atenção básica

Objetivo: Analisar a contribuição do enfermeiro para o aleitamento materno na atenção básica. Métodos: Trata-se de um estudo exploratório e descritivo, com abordagem qualitativa, realizado com 20 usuárias em período de lactação que estão cadastradas em uma Unidade de Saúde da Família do município de João Pessoa, Paraíba, Brasil. A coleta de dados foi realizada no domicílio das mulheres, mediante entrevista, utilizando um instrumento semiestruturado. As falas foram transcritas na íntegra e os dados foram analisados segundo a Técnica de Análise de Conteúdo. O estudo foi aprovado sob CAAE 65820217.7.00005176. Resultados: Emergiram duas categorias temáticas Contribuições do enfermeiro para a promoção do aleitamento materno durante o pré-natal e A visita puerperal como instrumento para a promoção do aleitamento materno. Conclusão: O enfermeiro apresenta um papel fundamental na orientação sobre ao aleitamento materno na atenção básica, desempenhando ações de promoção ainda durante o pré-natal e se estendendo até a visita puerperal

Nurse’s contribution to breastfeeding in basic attention / Contribuição do enfermeiro ao aleitamento materno na atenção básica

Objetivo: Analisar a contribuição do enfermeiro para o aleitamento materno na atenção básica. Métodos: Trata-se de um estudo exploratório e descritivo, com abordagem qualitativa, realizado com 20 usuárias em período de lactação que estão cadastradas em uma Unidade de Saúde da Família do município de João Pessoa, Paraíba, Brasil. A coleta de dados foi realizada no domicílio das mulheres, mediante entrevista, utilizando um instrumento semiestruturado. As falas foram transcritas na íntegra e os dados foram analisados segundo a Técnica de Análise de Conteúdo. O estudo foi aprovado sob CAAE 65820217.7.00005176. Resultados: Emergiram duas categorias temáticas Contribuições do enfermeiro para a promoção do aleitamento materno durante o pré-natal e A visita puerperal como instrumento para a promoção do aleitamento materno. Conclusão: O enfermeiro apresenta um papel fundamental na orientação sobre ao aleitamento materno na atenção básica, desempenhando ações de promoção ainda durante o pré-natal e se estendendo até a visita puerperal

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Minimal Symptom Expression' in Patients With Acetylcholine Receptor Antibody-Positive Refractory Generalized Myasthenia Gravis Treated With Eculizumab

The efficacy and tolerability of eculizumab were assessed in REGAIN, a 26-week, phase 3, randomized, double-blind, placebo-controlled study in anti-acetylcholine receptor antibody-positive (AChR+) refractory generalized myasthenia gravis (gMG), and its open-label extension

Post-intervention Status in Patients With Refractory Myasthenia Gravis Treated With Eculizumab During REGAIN and Its Open-Label Extension

OBJECTIVE: To evaluate whether eculizumab helps patients with anti-acetylcholine receptor-positive (AChR+) refractory generalized myasthenia gravis (gMG) achieve the Myasthenia Gravis Foundation of America (MGFA) post-intervention status of minimal manifestations (MM), we assessed patients' status throughout REGAIN (Safety and Efficacy of Eculizumab in AChR+ Refractory Generalized Myasthenia Gravis) and its open-label extension. METHODS: Patients who completed the REGAIN randomized controlled trial and continued into the open-label extension were included in this tertiary endpoint analysis. Patients were assessed for the MGFA post-intervention status of improved, unchanged, worse, MM, and pharmacologic remission at defined time points during REGAIN and through week 130 of the open-label study. RESULTS: A total of 117 patients completed REGAIN and continued into the open-label study (eculizumab/eculizumab: 56; placebo/eculizumab: 61). At week 26 of REGAIN, more eculizumab-treated patients than placebo-treated patients achieved a status of improved (60.7% vs 41.7%) or MM (25.0% vs 13.3%; common OR: 2.3; 95% CI: 1.1-4.5). After 130 weeks of eculizumab treatment, 88.0% of patients achieved improved status and 57.3% of patients achieved MM status. The safety profile of eculizumab was consistent with its known profile and no new safety signals were detected. CONCLUSION: Eculizumab led to rapid and sustained achievement of MM in patients with AChR+ refractory gMG. These findings support the use of eculizumab in this previously difficult-to-treat patient population. CLINICALTRIALSGOV IDENTIFIER: REGAIN, NCT01997229; REGAIN open-label extension, NCT02301624. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that, after 26 weeks of eculizumab treatment, 25.0% of adults with AChR+ refractory gMG achieved MM, compared with 13.3% who received placebo

ATLANTIC EPIPHYTES: a data set of vascular and non-vascular epiphyte plants and lichens from the Atlantic Forest

Epiphytes are hyper-diverse and one of the frequently undervalued life forms in plant surveys and biodiversity inventories. Epiphytes of the Atlantic Forest, one of the most endangered ecosystems in the world, have high endemism and radiated recently in the Pliocene. We aimed to (1) compile an extensive Atlantic Forest data set on vascular, non-vascular plants (including hemiepiphytes), and lichen epiphyte species occurrence and abundance; (2) describe the epiphyte distribution in the Atlantic Forest, in order to indicate future sampling efforts. Our work presents the first epiphyte data set with information on abundance and occurrence of epiphyte phorophyte species. All data compiled here come from three main sources provided by the authors: published sources (comprising peer-reviewed articles, books, and theses), unpublished data, and herbarium data. We compiled a data set composed of 2,095 species, from 89,270 holo/hemiepiphyte records, in the Atlantic Forest of Brazil, Argentina, Paraguay, and Uruguay, recorded from 1824 to early 2018. Most of the records were from qualitative data (occurrence only, 88%), well distributed throughout the Atlantic Forest. For quantitative records, the most common sampling method was individual trees (71%), followed by plot sampling (19%), and transect sampling (10%). Angiosperms (81%) were the most frequently registered group, and Bromeliaceae and Orchidaceae were the families with the greatest number of records (27,272 and 21,945, respectively). Ferns and Lycophytes presented fewer records than Angiosperms, and Polypodiaceae were the most recorded family, and more concentrated in the Southern and Southeastern regions. Data on non-vascular plants and lichens were scarce, with a few disjunct records concentrated in the Northeastern region of the Atlantic Forest. For all non-vascular plant records, Lejeuneaceae, a family of liverworts, was the most recorded family. We hope that our effort to organize scattered epiphyte data help advance the knowledge of epiphyte ecology, as well as our understanding of macroecological and biogeographical patterns in the Atlantic Forest. No copyright restrictions are associated with the data set. Please cite this Ecology Data Paper if the data are used in publication and teaching events. © 2019 The Authors. Ecology © 2019 The Ecological Society of Americ