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Increasing the fungicidal action of Amphotericin B by inhibiting the Nitric Oxi...

Vriens,Kim; Kumar, Phalguni Tewari; Struyfs, Caroline; Cools, Tanne L.; Spincemaille, Pieter; Kokalj, Tadej; Marques, Belém Sampaio; Ludovico, Paula

Amphotericin B (AmB) induces oxidative and nitrosative stresses, characterized by production of reactive oxygen and nitrogen species, in fungi. Yet, how these toxic species contribute to AmB-induced fungal cell death is unclear. We investigated the role of superoxide and nitric oxide radicals in AmB's fungicidal activity in Saccharomyces cerevisiae, using a digital microfluidic platform, which enabled monitorin...


Cell sheet engineering using the stromal vascular fraction of adipose tissue as...

Costa, M.; Cerqueira, Mariana Teixeira; Santos, T. C.; Marques, Belém Sampaio; Ludovico, Paula; Marques, A. P.; Pirraco, Rogério P.; Reis, R. L.

Current vascularization strategies for Tissue Engineering constructs, in particular cell sheet-based, are limited by time-consuming and expensive endothelial cell isolation and/or by the complexity of using extrinsic growth factors. Herein, we propose an alternative strategy using angiogenic cell sheets (CS) obtained from the stromal vascular fraction (SVF) of adipose tissue that can be incorporated into more c...


AMPK in Pathogens

Mesquita, Inês Morais; Moreira, Diana; Marques, Belém Sampaio; Laforge, Mireille; Cordeiro-da-Silva, Anabela; Ludovico, Paula; Estaquier, Jérôme

During host–pathogen interactions, a complex web of events is crucial for the outcome of infection. Pathogen recognition triggers powerful cellular signaling events that is translated into the induction and maintenance of innate and adaptive host immunity against infection. In opposition, pathogens employ active mechanisms to manipulate host cell regulatory pathways toward their proliferation and survival. Amon...


Proteolytic systems and AMP-activated protein kinase are critical targets of ac...

Fernandes, Ângela Margarida Alves; Azevedo, Maria M.; Pereira, Olga; Marques, Belém Sampaio; Paiva, Artur; Neves, Margarida Correia; Castro, Isabel

The therapeutic strategies against acute myeloid leukemia (AML) have hardly been modified over four decades. Although resulting in a favorable outcome in young patients, older individuals, the most affected population, do not respond adequately to therapy. Intriguingly, the mechanisms responsible for AML cells chemoresistance/ susceptibility are still elusive. Mounting evidence has shed light on the relevance o...


Sirtuins and proteolytic systems: implications for pathogenesis of synucleinopa...

Marques, Belém Sampaio; Ludovico, Paula

Insoluble and fibrillar forms of a-synuclein are the major components of Lewy bodies, a hallmark of several sporadic and inherited neurodegenerative diseases known as synucleinopathies. a-Synuclein is a natural unfolded and aggregation-prone protein that can be degraded by the ubiquitin-proteasomal system and the lysosomal degradation pathways. a-Synuclein is a target of the main cellular proteolytic systems, b...


Proteomic analysis of the action of the Mycobacterium ulcerans toxin mycolacton...

Gama, José B.; Ohlmeier, S.; Martins, Teresa G.; Fraga, Alexandra G.; Marques, Belém Sampaio; Carvalho, M. Alice; Proença, M. Fernanda R. P.

Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. The tissue damage characteristic of BU lesions is known to be driven by the secretion of the potent lipidic exotoxin mycolactone. However, the molecular action of mycolactone on host cell biology mediating cytopathogenesis is not fully understood. Here we applied two-dimensional electrophoresis (2-DE) to identify the mechanisms ...


Longevity pathways and maintenance of the proteome: the role of autophagy and m...

Marques, Belém Sampaio; Ludovico, Paula; Burhans, William C.

Ageing is a complex and multi-factorial process that results in the progressive accumulation of molecular alterations that disrupt different cellular functions. The budding yeast Saccharomyces cerevisiae is an important model organism that has significantly contributed to the identification of conserved molecular and cellular determinants of ageing. The nutrient-sensing pathways are well-recognized modulators o...


DNA replication stress-induced loss of reproductive capacity in S. cerevisiae a...

Weinberger, Martin; Marques, Belém Sampaio; Ludovico, Paula; Burhans, William C.

In many organisms, attenuation of growth signaling by caloric restriction or mutational inactivation of growth signaling pathways extends lifespan and protects against cancer and other age-related diseases. The focus of many efforts to understand these effects has been on the induction of oxidative stress defenses that inhibit cellular senescence and cell death. Here we show that in the model organism S. cerevi...


Involvement of yeast HSP90 isoforms in response to stress and cell death induce...

Silva, Maria Alexandra; Marques, Belém Sampaio; Fernandes, Ângela Margarida Oliveira; Carreto, Laura; Rodrigues, Fernando José dos Santos

Acetic acid-induced apoptosis in yeast is accompanied by an impairment of the general protein synthesis machinery, yet paradoxically also by the up-regulation of the two isoforms of the heat shock protein 90 (HSP90) chaperone family, Hsc82p and Hsp82p. Herein, we show that impairment of cap-dependent translation initiation induced by acetic acid is caused by the phosphorylation and inactivation of eIF2 alpha by...


The emerging role of autophagy/mitophagy in α-synuclein-induced toxicity: studi...

Marques, Belém Sampaio

α-Synuclein (α-syn) misfolding and aggregation is strongly associated with both idiopathic and familial forms of Parkinson disease (PD). Evidence suggests that α-syn has an impact on cell clearance routes and protein quality control systems, such as the ubiquitin-proteasome system (UPS) and autophagy. Recent advances in the key role of the autosomal recessive PARK2/PARKIN and PINK1 genes in mitophagy, highlight...


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