FM-IB-Artigos em Revistas Internacionais
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- Antimigratory and antiproliferative effects of the brain‐targeted peptide conjugate PepH3‐vCPP2319 on triple negative breast cancer cell culturesPublication . Gonçalves, Catarina; Castanho, Miguel A. R. B.; Cavaco, Marco; Neves, VeraTriple-negative breast cancer (TNBC) is an aggressive breast cancer subtype affecting mostly younger women with a poor 5-year overall survival. It is characterized by a high metastization rate, particularly to the brain, where the blood-brain barrier (BBB) hinders the pharmaceuticals delivery. New anticancer drugs able to inhibit cell migration are required to effectively prevent the development of metastasis. PepH3-vCPP2319 (AGILKRW(Ahx)WRRRYRRWRRRRRQRRRPRR-amide), consisting of the conjugation of the BBB peptide shuttle (BBBpS) PepH3 (AGILKRW-amide) to the anticancer peptide (ACP) vCPP2319 (WRRRYRRWRRRRRQRRRPRR-amide), was reported to have high anticancer activity (IC50 = 5.0 μM) toward highly aggressive TNBC cells (MDA-MB-231) paired with 2-fold increased accumulation in the brain when compared to unconjugated vCPP2319. Herein, we demonstrate that PepH3-vCPP2319 inhibits cell migration and proliferation in wound healing assays, outperforming the gold standard small chemical inhibitor, iCRT-3. The concentration required to inhibit cell migration is 10-fold lower for PepH3-vCPP2319 (0.5 μM) when compared with iCRT-3 (50 μM). Likewise, PepH3-vCPP2319 at 2.5 μM was more efficient in preventing cell proliferation when compared with 50 μM iCRT-3, with 45% reduction in spheroid diameter. This study sheds light on the antimetastatic potential of PepH3-vCPP2319 through abrogation of cell migration to distant sites, including the brain.
- Improved Triamcinolone acetonide-eluting contact lenses based on cyclodextrins and high hydrostatic pressure assisted complexationPublication . Marto-Costa, Carolina; Toffoletto, Nadia; Salema-Oom, Madalena; Antunes, Alexandra M. M.; Pinto, Carlos A.; Saraiva, Jorge A.; Silva-Herdade, Ana S.; Alvarez-Lorenzo, Carmen; Serro, AnaContact lenses (CLs) constitute an advantageous platform for the topical release of corticosteroids due to their prolonged contact with the eye. However, the lipophilic nature of corticosteroids hampers CLs' ability to release therapeutic amounts. Two approaches to improve loading and release of triamcinolone acetonide (TA) from poly(2-hydroxyethyl methacrylate)-based hydrogels were investigated: adding 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) to the monomers solution before polymerization (HEMA/i-CD) and an hydrogels' post-treatment with HP-β-CD (HEMA/p-CD). The effect of HP-β-CD and sterilization by high hydrostatic pressure (HHP) on the hydrogel properties (water content, oxygen and ion permeability, roughness, transmittance, and stiffness) was evaluated. The HEMA/i-CD hydrogels had stronger affinity for TA, sustaining its release for one day. HHP sterilization promoted the formation of cyclodextrin-TA complexes within the hydrogels, improving their drug-loading capacity »60 %. Cytotoxicity and irritability tests confirmed the safety of the therapeutic CLs. TA released from the hydrogels permeated through ocular tissues ex vivo and showed anti-inflammatory activity. Finally, a previously validated mathematical model was used to estimate the ability of the TA-loaded CLs to deliver therapeutic drug concentrations to the posterior part of the eye. Overall, HP-β-CD-containing CLs are promising candidates for the topical ocular application of TA as an alternative delivery system to intraocular injections.
- Therapeutic approach for Covid-19 patientsPublication . Saldanha, CarlotaThe biochemical effects of hydroxychloroquine, ivermectin, azithromycin molecules, and the zinc cation on functional properties of human erythrocyte will be presented. Among a wide range of therapeutically applications attributed to hydroxycloroquine the anti-inflammatory role will be herein highlight as well as for the azithromycin. The intervention of the ivermectin molecules into cell abortion mechanism of the virus replication will be described. The action of hydroxychloroquine, ivermectin, azithromycin molecules, and of zinc cation to prevent the spread and the replication SARS2-CoV2 Virus, as a first therapeutic approach for Covid-19 symptomatic patients, will be the aim of the present opinion.
- The role of the erythrocyte on humans with COVID-19Publication . Saldanha, CarlotaCOVID-19 is a new out-breaking human disease characterized by a severe acute respiratory syndrome caused by Coronavirus 2 (SARS-CoV-2). A pandemic state was swiftly declared by the World Health Organization due its fast transmissibility and huge human mortality index. After a brief introduction to the COVID -19 disease such as the ways of virus transportation inside the human body, the aim of this revision is to focus on the SARS-CoV2 infected erythrocyte properties which are considered biomarkers of COVID-19 patients and highlight the erythrocyte as a transporter for therapeutic drugs deliver in infected patients.
- Endothelial cell plasma membrane biomechanics mediates effects of pro-inflammatory factors on endothelial mechanosensors: vicious circle formation in atherogenic inflammationPublication . Barvitenko, Nadezhda; Ashrafuzzaman, Mohammad; Lawen, Alfons; Skverchinskaya, Elisaveta; Saldanha, Carlota; Manca, Alessia; Uras, Giuseppe; Aslam, Muhammad; Pantaleo, AntonellaChronic low-grade vascular inflammation and endothelial dysfunction significantly contribute to the pathogenesis of cardiovascular diseases. In endothelial cells (ECs), anti-inflammatory or pro-inflammatory signaling can be induced by different patterns of the fluid shear stress (SS) exerted by blood flow on ECs. Laminar blood flow with high magnitude is anti-inflammatory, while disturbed flow and laminar flow with low magnitude is pro-inflammatory. Endothelial mechanosensors are the key upstream signaling proteins in SS-induced pro- and anti-inflammatory responses. Being transmembrane proteins, mechanosensors, not only experience fluid SS but also become regulated by the biomechanical properties of the lipid bilayer and the cytoskeleton. We review the apparent effects of pro-inflammatory factors (hypoxia, oxidative stress, hypercholesterolemia, and cytokines) on the biomechanics of the lipid bilayer and the cytoskeleton. An analysis of the available data suggests that the formation of a vicious circle may occur, in which pro-inflammatory cytokines enhance and attenuate SS-induced pro-inflammatory and anti-inflammatory signaling, respectively.
- Contribution of technology to human cell propertiesPublication . Saldanha, CarlotaAfter explaining the meaning of SARS-CoV2, the protection rules for the disease caused by this virus are described in order to eradicate the resulting pandemic. Methods to differentiate asymptomatic from symptomatic patients will be mentioned. Human lungs, heart, kidney, endothelium and erythrocyte have specific binding sites for the SARS-CoV2. The aim of this opinion was to highlight some new disposable technology to identify two cell properties. One of them is the vascular endothelial cell (EC) receptor binding to the SARS-CoV2 and the other is related with red blood cells (RBCs) as SARS-CoV2 carrier.
- A colorimetric process to visualize erythrocyte exovesicles aggregatesPublication . Saldanha, Carlota; Santos, Nuno C.; Martins e Silva, JoãoA biochemistry laboratory class protocol is described in order to create an opportunity for students to apply by doing the theoretical concepts underlying biomolecules and vesicles properties, together with the principles of centrifugation and colorimetric methodologies. Through simple procedures the students will i) observe the segregation of the vesicles suspensions into two separate phases (phtalate esters gradient and vesicle aggregates); ii) visualize the vesicle aggregates by protein, enzyme, and phospholipids coloration processes; and iii) discuss and explain the visualized colors by proper biochemical reactions. The aims, objectives, methodology of teaching/learning, and assessment for this laboratory class are indicated.
- ATP modulates acute inflammation In Vivo through Dual Oxidase 1–derived H2O2 production and NF-κB activationPublication . De Oliveira, Sofia; López-Muñoz, Azucena; Candel, Sergio; Pelegrín, Pablo; Calado, Ângelo; Mulero, VictorianoDual oxidase 1 (Duox1) is the NADPH oxidase responsible for the H2O2 gradient formed in tissues after injury to trigger the early recruitment of leukocytes. Little is known about the signals that modulate H2O2 release from DUOX1 and whether the H2O2 gradient can orchestrate the inflammatory response in vivo. In this study, we report on a dominant-negative form of zebrafish Duox1 that is able to inhibit endogenous Duox1 activity, H2O2 release and leukocyte recruitment after tissue injury, with none of the side effects associated with morpholino-mediated Duox1 knockdown. Using this specific tool, we found that ATP release following tissue injury activates purinergic P2Y receptors, and modulates Duox1 activity through phospholipase C (PLC) and intracellular calcium signaling in vivo. Furthermore, Duox1-derived H2O2 is able to trigger the NF-κB inflammatory signaling pathway. These data reveal that extracellular ATP acting as an early danger signal is responsible for the activation of Duox1 via a P2YR/PLC/Ca(2+) signaling pathway and the production of H2O2, which, in turn, is able to modulate in vivo not only the early recruitment of leukocytes to the wound but also the inflammatory response through activation of the NF-κB signaling pathway.
- Pleiotropic and potentially beneficial effects of reactive oxygen species on the intracellular signaling pathways in endothelial cellsPublication . Barvitenko, Nadezhda; Skverchinskaya, Elisaveta; Lawen, Alfons; Matteucci, Elena; Saldanha, Carlota; Uras, Giuseppe; Manca, Alessia; Aslam, Muhammad; Pantaleo, AntonellaEndothelial cells (ECs) are exposed to molecular dioxygen and its derivative reactive oxygen species (ROS). ROS are now well established as important signaling messengers. Excessive production of ROS, however, results in oxidative stress, a significant contributor to the development of numerous diseases. Here, we analyze the experimental data and theoretical concepts concerning positive pro-survival effects of ROS on signaling pathways in endothelial cells (ECs). Our analysis of the available experimental data suggests possible positive roles of ROS in induction of pro-survival pathways, downstream of the Gi-protein-coupled receptors, which mimics insulin signaling and prevention or improvement of the endothelial dysfunction. It is, however, doubtful, whether ROS can contribute to the stabilization of the endothelial barrier.
- Long-term prognostic value of protein C activity, erythrocyte aggregation and membrane fluidity in transmural myocardial infarctionPublication . Sargento, Luis; Saldanha, Carlota; Monteiro, José; Perdigão, Carlota; Martins e Silva, JoãoThe objective of this study was to evaluate the long-term predictive value of the haemostatic, inflammatory and haemorheologic disturbances in transmural myocardial infarction (MI). Sixty-four (59 male) consecutive survivors of a MI, with a mean age of 58.3 +/- 12.0 years, were followed over a period of 36 months. Eighteen patients had a cardiovascular event defined as the composite of death, non-fatal MI, unstable angina and stroke. The haemostatic (protein C activity-PtC, antithrombin III, plasminogen activator inhibitor-1), haemorheologic (blood fluidity and components, erythrocyte membrane fluidity) and inflammatory (polymorphonuclear elastase, leukocyte count) profiles were determined at hospital discharge, using standard methodology. Our results can be summarized as follow: (i) at hospital discharge, the subgroup of patients with events had higher leukoactivity, leukocyte count, membrane fluidity, prognosis cyte count (7833.0 +/- 1696.0 vs. 10294.0 +/- 3129.0; p = 0.011), lower PtC (100.65 +/- 19.08 vs.81.25 +/- 19.95; p = 0.002), and lower erythrocyte aggregation (14.26 +/- 5.94 vs. 11.47 +/- 3.45; p = 0.031) in relation to the ones without events; (ii) By Cox regression the protein C activity lower tertile (OR 0.169; 0.045-0.628; p = 0.008); erythrocyte membrane outer layer fluidity upper tertile (OR 0.067; 95% CI 0.011 - 0.393; p = 0.003); and erythrocyte aggregation lower tertile (OR 0.182; 0.038 - 0.876; p = 0.034) were independent predictors of the composite endpoint. We can conclude that some haemostatic, haemorheologic and inflammatory disturbances, at hospital discharge, are long-term independent predictors of recurrent cardiovascular events in transmural myocardial infarction survivors.