Cathepsin D protects colorectal cancer cells from acetate-induced apoptosis through autophagy-independent degradation of damaged mitochondria

Acetate is a short-chain fatty acid secreted by Propionibacteria from the human intestine, known to induce mitochondrial apoptotic death in colorectal cancer (CRC) cells. We previously established that acetate also induces lysosome membrane permeabilization in CRC cells, associated with release of the lysosomal protease cathepsin D (CatD), which has a well-established role in the mitochondrial apoptotic cascade. Unexpectedly, we showed that CatD has an antiapoptotic role in this process, as pepstatin A (a CatD inhibitor) increased acetate-induced apoptosis. These results mimicked our previous data in the yeast system showing that acetic acid activates a mitochondria-dependent apoptosis process associated with vacuolar membrane permeabilization and release of the vacuolar protease Pep4p, ortholog of mammalian CatD. Indeed, this protease was required for cell survival in a manner dependent on its catalytic activity and for efficient mitochondrial degradation independently of autophagy. In this study, we therefore assessed the role of CatD in acetate-induced mitochondrial alterations. We found that, similar to acetic acid in yeast, acetate-induced apoptosis is not associated with autophagy induction in CRC cells. Moreover, inhibition of CatD with small interfering RNA or pepstatin A enhanced apoptosis associated with higher mitochondrial dysfunction and increased mitochondrial mass. This effect seems to be specific, as inhibition of CatB and CatL with E-64d had no effect, nor were these proteases significantly released to the cytosol during acetate-induced apoptosis. Using yeast cells, we further show that the role of Pep4p in mitochondrial degradation depends on its protease activity and is complemented by CatD, indicating that this mechanism is conserved. In summary, the clues provided by the yeast model unveiled a novel CatD function in the degradation of damaged mitochondria when autophagy is impaired, which protects CRC cells from acetate-induced apoptosis. CatD inhibitors could therefore enhance acetate-mediated cancer cell death, presenting a novel strategy for prevention or therapy of CRC.

Para cá e para lá dos muros: relações familiares na interface entre o interior e o exterior da prisão

Tese de Doutoramento em Sociologia

The role of Pep4p, the vacuolar yeast protease ortholog of human cathepsin D, in mitochondria-dependent apoptosis

Doctoral Thesis (PhD Programm on Molecular and Environmental Biology)

Characterization of phenolic compounds and antioxidant properties of Glycyrrhiza glabra L. rhizomes and roots

The present work aims to characterize and quantify the phenolic composition and to evaluate the antioxidant activity of Glycyrrhiza glabra L. (commonly known as licorice) rhizomes and roots. The antioxidant potential of its methanol/water extract could be related with flavones (mainly apigenin derivatives), flavanones (mainly liquirintin derivatives), a methylated isoflavone and a chalcone, identified in the extract. Lipid peroxidation inhibition was the most pronounced antioxidant effect (EC50=0.24±0.01 µg/mL and 22.74±2.42 µg/mL in TBARS and -carotene/linoleate assays, respectively), followed by free radicals scavenging activity (EC50=111.54±6.04 µg/mL) and, finally, reducing power (EC50=128.63±0.21 µg/mL). In this sense, licorice extract could be used as a source of antioxidants for pharmaceutical, cosmetic and/or food industries.

In vitro anti-Candida activity of Glycyrrhiza glabra L.

The severity and frequency of opportunistic fungal infections still growing, concomitantly to the increasing rates of antimicrobial drugs resistance. Natural matrices have been used over years due to its multitude of health benefits, including antifungal potential. Thus, the present work aims to evaluate the anti-Candida potential of the phenolic extract and individual phenolic compounds of Glycyrrhiza glabra L. (licorice), by disc diffusion assay, followed by determination of the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) for both planktonic cells and biofilms. Licorice extract evidenced inhibitory potential against the nineteen tested Candida strains, but no pronounced effect was observed by testing the most abundant individual phenolic compounds. Candida tropicalis strains were the most sensible, followed by Candida glabrata, Candida parapsilosis and, then, Candida albicans. Lower MIC and MFC values were achieved to C. glabrata and C. tropicalis, which confirms its susceptibility to licorice extract; however, for C. tropicalis strains a higher variability was observed. Anti-biofilm potential was also achieved, being most evident in some C. glabrata and C. tropicalis strains. In general, a twice concentration of the MIC was necessary for planktonic cells to obtain a similar potential to that one observed for biofilms. Thus, an upcoming approach for new antifungal agents, more effective and safer than the current ones, is stablished; notwithstanding, further studies are necessary in order to understand its mechanism of action, as also to assess kinetic parameters.

Production of secondary metabolites in cell suspension cultures of Ocimum sanctum L.

Dissertação de mestrado em Plant Molecular Biology, Biotechnology and Bioentrepreneurship

Chemical characterization and bioactive properties of Prunus avium L.: The widely studied fruits and the unexplored stems

The aim of this study was to characterize sweet cherry regarding nutritional composition of the fruits, and individual phytochemicals and bioactive properties of fruits and stems. The chromatographic profiles in sugars, organic acids, fatty acids, tocopherols and phenolic compounds were established. All the preparations (extracts, infusions and decoctions) obtained using stems revealed higher antioxidant potential than the fruits extract, which is certainly related with its higher phenolic compounds (phenolic acids and flavonoids) concentration. The fruits extract was the only one showing antitumor potential, revealing selectivity against HCT-15 (colon carcinoma) (GI50~74 μg/mL). This could be related with anthocyanins that were only found in fruits and not in stems. None of the preparations have shown hepatotoxicity against normal primary cells. Overall, this study reports innovative results regarding chemical and bioactive properties of sweet cherry stems, and confirmed the nutritional and antioxidant characteristics of their fruits.

Europe du Sud: Italie, Espagne et Portugal: priorité à l'éducation aux médias

(Excerto) Partant d'une perspective comparative de plusieurs recherches realisees en ltalie au cours des demieres annees sur les mineurs et Ia television, Elisa Manna, de Ia Fondation Censis souligne le besoin pour Ia recherche de rompre avec les hyperspecialisations academiques. Elle propose des pistes de comparaisons et de confrontations de methodes, de techniques et de resultats, sans jamais perdre de vue les recherches empiriques sur le terrain. II est impensable aujourd'hui, souligne Elisa Manna, de continuer a separer, d'un cote le monde institutionnel de Ia culture qui, prive des outils de connaissance necessaires, serait oblige d'agir a l'aveuglette, et d'autre part celui de Ia recherche academique et privee, fragmentee, divisee et frustree par l'impossibilite de faire valoir ses efforts d 'un point de vue operationnel.

Development of a poly(L-Lactic acid) based drug release system

Dissertação de mestrado em Biofísica e Bionanossistemas

De l'importance de la psychologie collective dans le sciences sociales: Serge Moscovici et les passions humaines

L'oeuvre de Moscovici se résume à une tentative de rapprochement du psychique et du social, ne serait-ce que parce que la rationalité et l'irrationalité sont transversales auz comportaments individuels et à l'activité sociale. En ce sens, l'individu n'est pas plus du ressort de la seule psychologie que la societé ne l'est de la seule sociologie. Simplifier la realité reviendrait à simplifier la connossaince de celle-ci. La psycho-sociologie de Moscovici nous invite donc à prendre en compte l'homme dans sa complétude, en tant que mixte de logique et de sentiments, de raison et de passions. On comprendra alors l'importance du paradigme écologique dans les sciences sociales comme forme de compréhension de ce qui relie l'homme à une entité plus vaste, le renvoyant aussi bien à la nature et à son ascendance animale qu'à la techinque et à sa destinée post-humaine.