In atherosclerotic lesions, vascular smooth muscle cells (VSMCs) represent half of the foam cell population, characterized by an aberrant accumulation of undigested lipids within lysosomes. Loss of lysosome function impacts VSMCs homeostasis and disease progression. Understanding the molecular mechanisms underlying lysosome dysfunction in these cells is, therefore, crucial. We identify cholesteryl hemiazelate (ChA), a stable oxidation end-product of cholesteryl-polyunsaturated fatty acid esters, as an inducer of lysosome malfunction in VSMCs. ChA-treated VSMCs acquire a foam cell-like phenotype, characterized by enlarged lysosomes full of ChA and neutral lipids. The lysosomes are perinuclear and exhibit degradative capacity and cargo exit defects. Lysosome luminal pH is also altered. Even though, the transcriptional response machinery and autophagy are not activated by ChA, the addition of recombinant lysosomal acid lipase (LAL) is able to rescue lysosome dysfunction. ChA significantly affects VSMCs proliferation and migration impacting atherosclerosis. In sum, this work shows that: 1) ChA is sufficient to induce lysosomal dysfunction in VSMCs; 2) In ChA-treated VSMCs, neither lysosome biogenesis nor autophagy are triggered; and 3) Recombinant LAL can be a therapeutic approach for lysosomal dysfunction.

MV repair can be performed in most patients undergoing aortic valve replacement. It should be the procedure of choice whenever feasible, because it is associated with lower early and late mortality rates and with freedom from reoperation in non-rheumatic patients.

Malaria remains a major world public health problem, contributing to poverty and inequality. It is urgent to find new efficacious tools with few adverse effects. Malaria has selected red blood cell (RBC) alterations linked to resistance against infection, and understanding the protective mechanisms involved may be useful for developing host-directed tools to control Plasmodium infection. Pyruvate kinase deficiency has been associated with resistance to malaria. Pyruvate kinase-deficient RBCs display an increased concentration of 2,3-diphosphoglycerate (2,3-DPG). We recently showed that 2,3-DPG impacts in vitro intraerythrocytic parasite growth, induces a shift of the metabolic profile of infected cells (iRBCs), making it closer to that of noninfected ones (niRBCs), and decreases the number of parasite progenies that invade new RBCs. As an increase of 2,3-DPG content may also have an adverse effect on RBC membrane and, consequently, on the parasite invasion, in this study, we explored modifications of the RBC morphology, biomechanical properties, and RBC membrane on Plasmodium falciparum in vitro cultures treated with 2,3-DPG, using atomic force microscopy (AFM)-based force spectroscopy and other experimental approaches. The presence of infection by P. falciparum significantly increased the rigidity of parasitized cells and influenced the morphology of RBCs, as parasitized cells showed a decrease of the area-to-volume ratio. The extracellular addition of 2,3-DPG also slightly affected the stiffness of niRBCs, making it more similar to that of infected cells. It also changed the niRBC height, making the cells appear more elongated. Moreover, 2,3-DPG treatment influenced the cell surface charge, becoming more negative in treated RBCs than in untreated ones. The results indicate that treatment with 2,3-DPG has only a mild effect on RBCs in comparison with the effect of the presence of the parasite on the host cell. 2,3-DPG is an endogenous host metabolite, which may, in the future, originate a new antimalarial tool with few adverse effects on noninfected cells.

Amyotrophic lateral sclerosis (ALS) is an aggressive neurodegenerative disorder related to neuroinflammation that is associated with increased risk of thrombosis. We aimed to evaluate γ' fibrinogen plasma level (an in vivo variant of fibrinogen) as a biomarker in ALS, and to test its role as a predictor of disease progression and survival. Sixty-seven consecutive patients with ALS were followed and the results were compared with those from 82 healthy blood donors. Patients were clinically evaluated at the time of blood sampling and on follow-up (every 3 months for the beginning of the follow-up until death) by applying the revised ALS Functional Rating Scale. Human plasma γ' fibrinogen concentration was quantified using a specific two-site sandwich kit enzyme-linked immunosorbent assay. We found, for the first time, a positive association between γ' fibrinogen concentration and survival in ALS patients: patients with higher γ' fibrinogen plasma levels survived longer, and this finding was not influenced by confounders such as age, gender, respiratory impairment, or functionality (ALSFRS-R score). Since increased levels have a positive impact on outcome, this novel biomarker should be further investigated in ALS.

Amyotrophic lateral sclerosis (ALS) is a devastating and fatal neurodegenerative disorder.Death typically occurs within 3–5 years after disease onset. The main cause of death in ALS is respiratory failure (RF). No effective treatment is available and no molecular biomarker related to respiratory outcome and to early ventilatory dysfunction was described so far. The club-cell protein (CC-16) is a biomarker associated with respiratory distress and lung inflammation. The aim of this work is to test if CC-16 and IL-6 could be new biomarkers of ALS for early signs of respiratory insufficiency and disease progression. Additionally, we intend to study morphological and viscoelastic changes of the erythrocytes’ membrane associating them with ALS patients’ clinical profile. Eighty-one ALS patients and 30 matched controls were included. Functional capacity and respiratory function (forced vital capacity) were evaluated. CC-16 and IL-6 were quantified by ELISA and multiplex technology, respectively. Morphological and viscoelastic properties of the erythrocytes were analysed by Atomic Force Microscopy (AFM). CC-16 levels were significantly raised in ALS patients. In 17% of them, CC-16 level was above the upper cut-off value. On these patients, the risk of non-invasive ventilation was greater in the following 6 months and they tend to have higher mortality in the following 30 months. IL-6 values were not different in ALS population as compared with controls. ALS patients have higher erythrocyte maximum height, area and volume, decreased erythrocyte membrane roughness and increased membrane stiffness than the control group. These results indicates abnormal erythrocyte structure and possible changes on membrane lipid composition on ALS patients. We propose that increased CC-16 levels could be a marker of lung inflammatory response, associated with ventilatory insufficiency and related to impending respiratory failure, which are not fully predicted by conventional respiratory tests. Moreover, abnormalities in erythrocyte morphology may enhance the risk of tissue hypoxia. Amyotrophic lateral sclerosis (ALS) is a devastating and fatal neurodegenerative disorder. Death typically occurs within 3–5 years after disease onset. Respiratory failure (RF) is the main cause of death. No effective treatment and no molecular biomarker related to respiratory outcome and to early ventilatory dysfunction are available. The club-cell protein (CC-16) is a biomarker associated with respiratory distress and lung inflammation. The aim of this work is to test if CC-16 and IL-6 could be new biomarkers of ALS for early signs of respiratory insufficiency and disease progression. Additionally, we intend to study morphological and viscoelastic changes of the erythrocytes’ membrane associating them with ALS patients’ clinical profile. Functional capacity and respiratory function (forced vital capacity) were evaluated in 81 ALS patients and 30 ma...