The adoption of sustainable practices on personal luxury goods : impact on purchase motivations and intention, emotions, and self-identity

With the growth of the luxury industry in recent years, especially the personal goods segment, and with environmental concerns and sustainability increasingly influencing consumers, it is crucial to understand the role of this industry in the spotlight of sustainability. For this reason, this investigation aims to understand what perceptions, feelings, and purchase intentions current and potential consumers will have about these sustainable luxury products. To do this, based on the literature, information was gathered from individuals by (1) developing a questionnaire to assess consumers’ perceptions and attitudes towards different sustainable strategies, and (2) conduction interviews to deepen the knowledge regarding consumer’s emotions regarding sustainable luxury products. This way, it was possible to assess people's perspectives and understanding of the new sustainable production methods that have been implemented. The results of this investigation reveal that, in general, individuals are considerably concerned about the current state of the environment and that despite their positive purchase intentions for this type of sustainable goods, perceptions about them are not unanimous and deserve further look. Thus, this study provides relevant academic and managerial contributions, as it indicates which sustainable strategies are most familiar to individuals, how they judge and evaluate them, and which of them generates higher purchase intentions, granting a deeper knowledge about the potential consumers in this market segment.Com o crescimento da indústria do luxo nos últimos anos, especialmente no segmento de bens pessoais, e com a sustentabilidade e preocupações ambientais a influenciarem cada vez mais os consumidores, torna-se crucial compreender o papel desta indústria num mundo que caminha para um futuro que se pretende cada vez mais sustentável. Por esta razão, esta investigação visa compreender que perceções, sentimentos e intenções de compra terão os consumidores atuais e potenciais sobre estes produtos de luxo sustentáveis. Para tal, com base na literatura, foram recolhidas informações junto de indivíduos por meio de (1) desenvolvimento de um questionário para avaliar as perceções e atitudes dos consumidores em relação a diferentes estratégias sustentáveis e (2) realização de entrevistas para aprofundar as emoções dos mesmos em relação aos produtos de luxo sustentáveis. Desta forma, foi possível avaliar a perspetiva e compreensão das pessoassobre os novos métodos de produção sustentável que têm vindo a ser implementados. Os resultados desta investigação revelam que, de uma forma geral, os indivíduos estão bastante preocupados com o estado atual do ambiente e que apesar das intenções de compra perante este tipo de bens ser positiva, as perceções sobre os mesmos não são unânimes e merecem ser aprofundadas. Assim, este estudo fornece contributos académicos relevantes para a gestão, visto que indica quais estratégias sustentáveis são mais familiares aos indivíduos, como estes as avaliam e quais geram maiores intenções de compra, proporcionando um conhecimento mais profundo sobre os potenciais consumidores deste segmento de mercado

Metabolic cooperation between cancer and non-cancerous stromal cells is pivotal in cancer progression

The way cancer cells adapt to microenvironment is crucial for the success of carcinogenesis, and metabolic fitness is essential for a cancer cell to survive and proliferate in a certain organ/tissue. The metabolic remodeling in a tumor niche is endured not only by cancer cells but also by non-cancerous cells that share the same microenvironment. For this reason, tumor cells and stromal cells constitute a complex network of signal and organic compound transfer that supports cellular viability and proliferation. The intensive dual-address cooperation of all components of a tumor sustains disease progression and metastasis. Herein, we will detail the role of cancer-associated fibroblasts, cancer-associated adipocytes, and inflammatory cells, mainly monocytes/macrophages (tumor-associated macrophages), in the remodeling and metabolic adaptation of tumors.publishersversionpublishe

Role of fibronectin in tumor development and progression

Tese de mestrado. Biologia (Biologia Humana e Ambiente). Universidade de Lisboa, Faculdade de Ciências, 2011A tumorigénese é um processo complexo cujas várias fases reflectem alterações genéticas que resultam na transformação progressiva de células normais em células malignas (Hanahan & Weinberg, 2000; Renan, 1993). As principais características que ditam colectivamente o cancro são: auto-suficiência em relação aos sinais de crescimento, insensibilidade aos sinais de anti-crescimento, evasão à apoptose, elevado potencial replicativo, angiogénese sustentada, evasão ao controlo imunitário, reprogramação metabólica, invasão tecidular e metastização, representando esta última a principal causa de morbidez e morte associadas à doença (Hanahan & Weinberg, 2011). Todos estes atributos resultam de uma interacção dinâmica entre as células tumorais e o seu microambiente – microambiente tumoral –, que inclui células normais, factores mediadores e componentes da matriz extracelular (MEC) (Tlsty & Coussens, 2006). A fibronectina (FN) é uma glicoproteína modular de elevado peso molecular que pode ser classificada, de acordo com a sua solubilidade, em FN plasmática, solúvel, e FN celular, uma forma menos solúvel que representa o maior componente da MEC (Klein et al, 2004; Pankov & Yamada, 2002). Estruturalmente, a FN apresenta-se sob a forma de um dímero composto por duas cadeias polipeptídicas aproximadamente idênticas, ligadas covalentemente por um par de pontes dissulfito próximo da extremidade C-terminal. Cada monómero tem um peso molecular de 230-250 kDa e é composto por três tipos de unidades de repetição ou módulos: o tipo I (FI), o tipo II (FII) e o tipo III (FIII) (Hynes & Yamada, 1982; Pankov & Yamada, 2002; Potts & Campbell, 1996). A FN, que tem como receptores integrinas, está implicada numa grande variedade de funções celulares, sobretudo envolvendo interacções entre as células e a MEC, onde assume grande importância na adesão, morfologia, migração, crescimento e diferenciação celulares, organização do citosqueleto e hemostasia (Hynes & Yamada, 1982; Kornblihtt et al, 1996; Labat-Robert, 2002). As interacções célula-célula e célula-matriz são essenciais para o fornecimento de informações que regulam o normal funcionamento celular, pelo que a degradação ou activação das proteínas de matriz e/ou da superfície celular podem mediar alterações irreversíveis no microambiente celular (Werb, 1997). O crescimento tumoral, angiogénese, invasão e metastização estão fortemente dependentes da natureza permissiva do microambiente. Durante estas fases, a proteólise da MEC representa um passo crucial (Chambers & Matrisian, 1997; Polette et al, 2004), sendo reconhecida à FN uma forte susceptibilidade neste âmbito (Kenny et al, 2008). As metaloproteases de matriz (MPMs) constituem a família de proteases mais importante associada à tumorigénese (Kessenbrock et al, 2010). As MPM-9 e MPM-2 têm sido apontadas como as MPMs mais importantes na metastização (Coussens et al, 2002; Klein et al, 2004; Malemud, 2006; Stetler-Stevenson, 1999). O aumento concomitante de FN e enzimas proteolíticas, bem como de fragmentos de fibronectina tem sido observado em doentes de cancro (Katayama et al, 1993; Kenny et al, 2008; Kenny & Lengyel, 2009; Labat-Robert, 2002; Labat-Robert et al, 1980; Warawdekar et al, 2006), sugerindo uma possível intervenção da FN e seus fragmentos no processo tumoral, embora o eventual mecanismo subjacente ao fenómeno não seja ainda claro. Paralelamente, outros estudos têm atribuído propriedades anti-tumorais a alguns fragmentos derivados da FN (Humphries et al, 1986; Humphries et al, 1988; Kato et al, 2002; Saiki, 1997; Yi & Ruoslahti, 2001). Com base no trabalho científico que vem sendo dedicado a esta temática, foi estabelecido como objectivo deste trabalho o estudo do papel da FN no desenvolvimento e progressão tumorais. Com este intuito, foi desenvolvido um trabalho experimental que partiu do estabelecimento de dois grupos, para diferentes linhas celulares, diferindo entre si nos níveis de FN produzida. Para isso, procedeu-se à construção de um plasmídeo contendo o gene da FN embrionária (full-length), pcDNA3-flFN. Este vector foi transfectado em células das linhas celulares tumorais humanas HCT15 (carcinoma colorrectal) e HeLa (adenocarcinoma do colo do útero), e da linha celular não-tumoral CHO, uma linha celular transformada onde o mecanismo de controlo de transcrição se encontra inactivado, tendo sido, por isso, utilizada como controlo positivo, in vitro. Para cada linha celular foi mantido um grupo de células controlo, não transfectadas. O trabalho experimental consistiu no desenvolvimento de ensaios in vitro e in vivo, após a indução da formação de tumores xenógrafos em modelos ortotópicos de ratinhos BALB/c-SCID. Os resultados foram comparados entre grupos. Após transfecção estável das linhas celulares, a expressão de FN foi determinada, para cada grupo, por PCR quantitativo em tempo-real. A detecção de níveis mais elevados de mRNA nos grupos controlo, para todas as linhas celulares – sobretudo em HCT15 e CHO –, validou o sucesso da transfecção, o que permitiu a progressão do trabalho experimental. Análises de western blotting – foram utilizadas como amostras a FN imunoprecipitada a partir do meio de cultura das células – e imunofluorescência confirmaram a existência de uma correlação entre níveis de mRNA e proteína, evidenciando valores proteicos de FN aumentados nos grupos transfectados, em comparação com os grupos controlo. Os resultados obtidos por western blotting mostraram, adicionalmente, a presença de fragmentos proteolíticos de FN nas linhas tumorais, não observada no controlo positivo (FN imunoprecipitada a partir de soro de um individuo saudável), o que está de acordo com as observações de vários estudos realizados em doentes de cancro (Katayama et al, 1993; Kenny et al, 2008; Kenny & Lengyel, 2009; Labat-Robert, 2002; Labat-Robert et al, 1980; Warawdekar et al, 2006). Para as linhas celulares CHO e HCT15, foram detectados níveis mais elevados de fragmentos proteolíticos de FN nos grupos transfectados, o que está de acordo com o esperado, dada a maior concentração de FN nestes grupos. Por imunofluorescência, foi observada, globalmente, uma maior expressão de integrinas nas células transfectadas, resultado esperado sendo essas os receptores da FN. Por zimografia, foi avaliada a actividade das MPMs. Foi detectada actividade proteolítica das MPM-9 e MPM-2, consideradas as MPMs mais importantes no processo de metastização (Coussens et al, 2002; Klein et al, 2004; Malemud, 2006; Stetler-Stevenson, 1999). Para a MPM-9 foi observada uma actividade proteolítica aumentada nos grupos transfectados de HCT15 e CHO, assim como para a MPM-2 em HCT15. Estes resultados podem estar relacionados com a maior concentração de FN nos meios de cultura das células transfectadas e revelaram-se também consistentes com o padrão de fragmentos de FN obtido por western blotting. O estudo da migração celular foi abordado por wound healing assay, do qual resultaram taxas de migração direccional mais elevadas para as células CHO e HCT15 transfectadas, facto que poderá estar relacionado com os resultados obtidos na zimografia. Um eventual envolvimento da FN na angiogénese foi avaliado por tube formation assay com HUVEC (células endoteliais de cordão umbilical humano) e pela quantificação da expressão, a nível de mRNA, de três isoformas de VEGF (factor de crescimento do endotélio vascular), receptores do VEGF e angiopoietinas. Os resultados mostraram uma relação inversa entre os níveis de FN e a formação de tubos. Colectivamente, os resultados in vitro sugeriram uma correlação entre níveis mais elevados de FN e um comportamento mais agressivo por parte das células, num contexto neoplásico, sugerindo um particular envolvimento da migração das células tumorais. Após aproximadamente dois meses da indução da formação de tumores nos modelos BALB/c-SCID, os animais foram sacrificados e procedeu-se à colheita dos principais órgão-alvo de metastização. A análise histológica dos tumores e órgãos recolhidos revelou mais invasão e metastização nos grupos inoculados com células controlo. Estas observações foram confirmadas para o grupo inoculado com HCT15, pela detecção de mais mastócitos nos tumores, os quais estão implicados no crescimento tumoral e invasão (Strouch et al, 2010), e pela quantificação da expressão de SCF (factor de células estaminais) – citocina envolvida na promoção da sobrevivência, proliferação e migração de mastócitos e células progenitoras (Broudy, 1997) – e SDF-1 (factor derivado do estroma da medula óssea) – citocina associada ao crescimento tumoral, angiogénese, metastização e recrutamento de células derivadas da medula óssea (Kryczek et al, 2007) –, que se revelou menor nas células transfectadas. Os resultados in vivo evidenciam, deste modo, um processo tumorigénico menos avançado nos animais inoculados com células transfectadas, contrariando as observações in vitro. Com efeito, uma taxa migratória mais elevada não implica necessariamente uma maior actividade invasiva por parte das céluals tumorais, a principal responsável pela disseminação do cancro. Por outro lado, as condições in vitro não incluem muitas variáveis existentes in vivo. Recomenda-se, no entanto, a validação dos modelos in vivo sob condições técnicas mais apropriadas. Não obstante, parece relevante a sequenciação de um fragmento detectado, para posterior investigação.Reciprocal interactions among normal cells, their mediators, components of the extracellular matrix (ECM) and genetically altered cells regulate all aspects of tumorigenicity. Fibronectin (FN), a multidomain glycoprotein, represents the major component of ECM and is implicated in a variety of cell functions, particularly those involving interactions between cells and ECM. ECM proteolysis is a crucial step during cancer stages and FN strong susceptibility to proteolytic degradation is well documented. Indeed, several studies have related FN levels to tumor progression in cancer patients, observing an increase of both FN and FN fragments levels. In parallel, some research has, on the other hand, provided evidence of protective roles of fragments derived from FN. Based on the literature that has been dedicated to the subject, it was established as objective of this work the study of the role of FN in tumor development and progression. For this purpose, an experimental work was developed arising from the establishment of two groups within each cell line used – tumor cell lines HCT15 and HeLa, and CHO –, differing in the levels of FN produced. In vitro and in vivo assays were performed and the results compared between groups. Our findings suggested a correlation between higher FN levels and a more aggressive behavior of cells in a neoplastic context, in vitro. In vivo, the opposite was observed. It is recommended, however, the validation of the in vivo models under improved technical conditions. Nevertheless, the sequence of a fragment detected appeared to be relevant for further investigation

A Metabolomics-Inspired Strategy for the Identification of Protein Covalent Modifications

This work was supported by Fundação para a Ciência e a Tecnologia (FCT), Portugal, through projects UID/QUI/00100/2019, IF/01091/2013/CP1163/CT0001 and PTDC/QUIQAN/32242/2017 as well as doctoral fellowships SFRH/BD/102846/2014 (to CC) and SFRH/BD/140157/2018 (to JN);joint funding from FCT and the COMPETE Program is also acknowledge through RNEM-LISBOA-01-0145-FEDER-022125-funded postdoctoral fellowship (to JM).Identification of protein covalent modifications (adducts) is a challenging task mainly due to the lack of data processing approaches for adductomics studies. Despite the huge technological advances in mass spectrometry (MS) instrumentation and bioinformatics tools for proteomics studies, these methodologies have very limited success on the identification of low abundant protein adducts. Herein we report a novel strategy inspired on the metabolomics workflows for the identification of covalently-modified peptides that consists on LC-MS data preprocessing followed by statistical analysis. The usefulness of this strategy was evaluated using experimental LC-MS data of histones isolated from HepG2 and THLE2 cells exposed to the chemical carcinogen glycidamide. LC-MS data was preprocessed using the open-source software MZmine and potential adducts were selected based on the m/z increments corresponding to glycidamide incorporation. Then, statistical analysis was applied to reveal the potential adducts as those ions are differently present in cells exposed and not exposed to glycidamide. The results were compared with the ones obtained upon the standard proteomics methodology, which relies on producing comprehensive MS/MS data by data dependent acquisition and analysis with proteomics data search engines. Our novel strategy was able to differentiate HepG2 and THLE2 and to identify adducts that were not detected by the standard methodology of adductomics. Thus, this metabolomics driven approach in adductomics will not only open new opportunities for the identification of protein epigenetic modifications, but also adducts formed by endogenous and exogenous exposure to chemical agents.publishersversionpublishe

Propranolol therapy for cerebral cavernous malformations

Funding Information: The present study was funded by IPOLFG EPE and by iNOVA4Health (UID/Multi/04462/2019) a program finan‑ cially supported by Fundação para a Ciência e Tecnologia (FCT)/Ministério da Educação e Ciência, through national funds. The PhD fellowship of FLC was funded by FCT (PD/BD/128337/2017).Cerebral cavernous malformations (CCMs) are vascular malformations characterized by the abnormal growth of vascular structures in the central nervous system. However, the precise mechanism(s) responsible for the development of CCM vascular abnormalities remain poorly understood. Although the mechanisms of action of propranolol in CCM have not yet been fully explored it is not commonly prescribed, it has been shown to be effective in children and appears to play a protective role in the prevention of CCM-derived hemorrhage in adults. The present study performed in vitro and ex vivo assays in order to examine the effects of propranolol on endothelial cells (ECs). The percentage of CD14+/CD31+ cells and the levels of VEGF in the peripheral blood (PB) of a child patient with CCM, with recurrent seizures and hemorrhages, who was maintained under propranolol therapy, were also analyzed. In addition to the effects of propranolol on differentiated ECs, and the decrease angiogenic-related features in vitro and ex vivo, it was observed that in the PB of this patient, propranolol administration decreased the percentage of circulating cells sharing monocytic and EC features (CD14+/CD31+ cells), as well as the VEGF levels; this was concomitant with a good prognosis and with the reversion of CCM lesions. A decrease in VEGF levels by propranolol may also be involved in the impairment of the recruitment of CD14+/CD31+ monocytes functioning as endothelial progenitor cells to sustain the vascular lesion. On the whole, the present study demonstrates that propranolol impairs angiogenesis in vitro and may thus be a useful tool for the clinical management of CCM. Moreover, the present study highlights the monitorization of the levels of CD14+/CD31+ monocytes and VEGF levels as a useful tool for predicting the clinical efficacy of propranolol in patients with CCM.publishersversionepub_ahead_of_prin

Monocytes as Endothelial Progenitor Cells (EPCs), Another Brick in the Wall to Disentangle Tumor Angiogenesis

The project was funded by IPOLFG, EPE, by iNOVA4Health (UID/Multi/04462/2019) a program financially supported by Fundação para a Ciência e Tecnologia/Ministério da Educação e Ciência, through national funds and co-funded by FEDER under the PT2020 Partnership Agreement and by Fundação para a Ciência eTecnologia (PhD student fellowship: PD/BD/128337/2017).Bone marrow contains endothelial progenitor cells (EPCs) that, upon pro-angiogenic stimuli, migrate and differentiate into endothelial cells (ECs) and contribute to re-endothelialization and neo-vascularization. There are currently no reliable markers to characterize EPCs, leading to their inaccurate identification. In the past, we showed that, in a panel of tumors, some cells on the vessel wall co-expressed CD14 (monocytic marker) and CD31 (EC marker), indicating a putative differentiation route of monocytes into ECs. Herein, we disclosed monocytes as potential EPCs, using in vitro and in vivo models, and also addressed the cancer context. Monocytes acquired the capacity to express ECs markers and were able to be incorporated into blood vessels, contributing to cancer progression, by being incorporated in tumor neo-vasculature. Reactive oxygen species (ROS) push monocytes to EC differentiation, and this phenotype is reverted by cysteine (a scavenger and precursor of glutathione), which indicates that angiogenesis is controlled by the interplay between the oxidative stress and the scavenging capacity of the tumor microenvironment.publishersversionpublishe

Cysteine allows ovarian cancer cells to adapt to hypoxia and to escape from carboplatin cytotoxicity

The authors would like to acknowledge the Instituto Portugues de Oncologia de Lisboa Francisco Gentil (IPOLFG) for partially funding the project. We would also like to acknowledge Dr Dialina Brilhante and Dr. Teresa Guerreiro (Servico de Imuno-hemoterapia, IPOLFG) for providing blood donors samples; to Dr Humberto Goncalves (Pharmacy, IPOLFG) for paclitaxel and carboplatin preparation, and Marta Teixeira (IBET) for the technical support in 3D models. The study was also funded by Projecto TVI. This research was supported by Fundacao para a Ciencia e Tecnologia (FCT) (PhD ProRegeM program, PD/BD/105893/2014, FCT fellowship, PD/BD/105768/2014). iNOVA4Health - UID/Multi/04462/2013, a program financially supported by Fundacao para a Ciencia e Tecnologia/Ministerio da Educacao e Ciencia, through national funds and co-funded by FEDER under the PT2020 Partnership Agreement is acknowledged.Ovarian cancer is the second most common gynaecologic malignancy and the main cause of death from gynaecologic cancer, due to late diagnosis and chemoresistance. Studies have reported the role of cysteine in cancer, by contributing for hydrogen sulphide (H2S) generation and as a precursor of glutathione (GSH). However, the role of cysteine in the adaptation to hypoxia and therapy response remains unclear. We used several ovarian cancer cell lines, ES2, OVCAR3, OVCAR8, A2780 and A2780cisR, to clarify cysteine relevance in ovarian cancer cells survival upon hypoxia and carboplatin. Results show that ES2 and OVCAR8 cells presented a stronger dependence on cysteine availability upon hypoxia and carboplatin exposure than OVCAR3 cells. Interestingly, the A2780 cisR, but not A2780 parental cells, benefits from cysteine upon carboplatin exposure, showing that cysteine is crucial for chemoresistance. Moreover, GSH degradation and subsequent cysteine recycling pathway is associated with ovarian cancer as seen in peripheral blood serum from patients. Higher levels of total free cysteine (Cys) and homocysteine (HCys) were found in ovarian cancer patients in comparison with benign tumours and lower levels of GSH were found in ovarian neoplasms patients in comparison with healthy individuals. Importantly, the total and S-Homocysteinylated levels distinguished blood donors from patients with neoplasms as well as patients with benign from patients with malignant tumours. The levels of S-cysteinylated proteins distinguish blood donors from patients with neoplasms and the free levels of Cys in serum distinguish blood from patients with benign tumours from patients with malignant tumours. Herein we disclosed that cysteine contributes for a worse disease prognosis, allowing faster adaptation to hypoxia and protecting cells from carboplatin. The measurement of serum cysteine levels can be an effective tool for early diagnosis, for outcome prediction and follow up of disease progression.publishersversionpublishe

EuReCa ONE—27 Nations, ONE Europe, ONE Registry A prospective one month analysis of out-of-hospital cardiac arrest outcomes in 27 countries in Europe

AbstractIntroductionThe aim of the EuReCa ONE study was to determine the incidence, process, and outcome for out of hospital cardiac arrest (OHCA) throughout Europe.MethodsThis was an international, prospective, multi-centre one-month study. Patients who suffered an OHCA during October 2014 who were attended and/or treated by an Emergency Medical Service (EMS) were eligible for inclusion in the study. Data were extracted from national, regional or local registries.ResultsData on 10,682 confirmed OHCAs from 248 regions in 27 countries, covering an estimated population of 174 million. In 7146 (66%) cases, CPR was started by a bystander or by the EMS. The incidence of CPR attempts ranged from 19.0 to 104.0 per 100,000 population per year. 1735 had ROSC on arrival at hospital (25.2%), Overall, 662/6414 (10.3%) in all cases with CPR attempted survived for at least 30 days or to hospital discharge.ConclusionThe results of EuReCa ONE highlight that OHCA is still a major public health problem accounting for a substantial number of deaths in Europe.EuReCa ONE very clearly demonstrates marked differences in the processes for data collection and reported outcomes following OHCA all over Europe. Using these data and analyses, different countries, regions, systems, and concepts can benchmark themselves and may learn from each other to further improve survival following one of our major health care events