BACKGROUND: Growing evidence correlated changes in bioactive sphingolipids, particularly S1P (sphingosine-1-phosphate) and ceramides, with coronary artery diseases. Furthermore, specific plasma ceramide species can predict major cardiovascular events. Dysfunction of the endothelium lining lesion-prone areas plays a pivotal role in atherosclerosis. Yet, how sphingolipid metabolism and signaling change and contribute to endothelial dysfunction and atherosclerosis remain poorly understood. METHODS: We used an established model of coronary atherosclerosis in mice, combined with sphingolipidomics, RNA-sequencing, flow cytometry, and immunostaining to investigate the contribution of sphingolipid metabolism and signaling to endothelial cell (EC) activation and dysfunction. RESULTS: We demonstrated that hemodynamic stress induced an early metabolic rewiring towards endothelial sphingolipid de novo biosynthesis, favoring S1P signaling over ceramides as a protective response. This finding is a paradigm shift from the current belief that ceramide accrual contributes to endothelial dysfunction. The enzyme SPT (serine palmitoyltransferase) commences de novo biosynthesis of sphingolipids and is inhibited by NOGO-B (reticulon-4B), an ER membrane protein. Here, we showed that NOGO-B is upregulated by hemodynamic stress in myocardial EC of ApoE ^−/− mice and is expressed in the endothelium lining coronary lesions in mice and humans. We demonstrated that mice lacking NOGO-B specifically in EC (Nogo-A/B ^ECKO ApoE ^−/− ) were resistant to coronary atherosclerosis development and progression, and mortality. Fibrous cap thickness was significantly increased in Nogo-A/B ^ECKO ApoE ^−/− mice and correlated with reduced necrotic core and macrophage infiltration. Mechanistically, the deletion of NOGO-B in EC sustained the rewiring of sphingolipid metabolism towards S1P, imparting an atheroprotective endothelial transcriptional signature. CONCLUSIONS: These data demonstrated that hemodynamic stress induced a protective rewiring of sphingolipid metabolism, favoring S1P over ceramide. NOGO-B deletion sustained the rewiring of sphingolipid metabolism toward S1P protecting EC from activation under hemodynamic stress and refraining coronary atherosclerosis. These findings also set forth the foundation for sphingolipid-based therapeutics to limit atheroprogression.

Breast cancer is the most frequent form of cancer occurring in women of any age. Among the different types, the triple-negative breast cancer (TNBC) subtype is recognized as the most severe form, being associated with the highest mortality rate. Currently, there are no effective treatments for TNBC. For this reason, the research of novel therapeutics is urgently needed. Natural products and their analogs have historically made a major contribution to pharmacotherapy and the treatment of various human diseases, including cancer. In this study, we explored the potential anti-cancer effects of erucin, the most abundant H₂S-releasing isothiocyanate present in arugula (Eruca sativa) in MDA-MB-231 cells, a validated in vitro model of TNBC. We found that erucin, in a concentration-dependent manner, significantly inhibited MDA-MB-231 cell proliferation by inducing apoptosis and autophagy. Additionally, erucin prevented intracellular ROS generation promoting the expression of key antioxidant genes and halted MDA-MB-231 cell migration, invasion, and colony formation. In conclusion, using a cellular and molecular biology approach, we show that the consumption of erucin could represent a novel and promising strategy for intervention against TNBC.

Thrombin binding aptamer (TBA) is one of the best-known G-quadruplex (G4)-forming aptamers that efficiently binds to thrombin, resulting in anticoagulant effects. TBA also possesses promising antiproliferative properties. As with most therapeutic oligonucleotides, chemical modifications are critical for therapeutic applications, particularly to improve thermodynamic stability, resistance in biological environment, and target affinity. To evaluate the effects of nucleobase and/or sugar moiety chemical modifications, five TBA analogues have been designed and synthesized considering that the chair-like G4 structure is crucial for biological activity. Their structural and biological properties have been investigated by Circular Dichroism (CD), Nuclear Magnetic Resonance (NMR), native polyacrylamide gel electrophoresis (PAGE) techniques, and PT and MTT assays. The analogue TBAB contains 8-bromo-2′-deoxyguanosine (B) in G-syn glycosidic positions, while TBAL and TBAM contain locked nucleic acid guanosine (L) or 2′-O-methylguanosine (M) in G-anti positions, respectively. Instead, both the two types of modifications have been introduced in TBABL and TBABM with the aim of obtaining synergistic effects. In fact, both derivatives include B in syn positions, exhibiting in turn L and M in the anti ones. The most appealing results have been obtained for TBABM, which revealed an interesting cytotoxic activity against breast and prostate cancer cell lines, while in the case of TBAB, extraordinary thermal stability (T_m approximately 30 °C higher than that of TBA) and an anticoagulant activity higher than original aptamer were observed, as expected. These data indicate TBAB as the best TBA anticoagulant analogue here investigated and TBABM as a promising antiproliferative derivative.

Nitric oxide (NO) is a pivotal mediator of numerous physiological and pathological events, with its reactive derivatives, nitrite and nitrate, reflecting both cellular signaling and stress conditions. Among these, nitrite is widely recognised as a stable surrogate marker of NO production, and its accumulation is associated with inflammation, cardiovascular dysfunction, and tumor progression. Conventional quantification relies on the Griess assay, which, although reliable, requires multiple reagents, extended handling, and spectrophotometric instrumentation. Here, we present a simple and inexpensive electrochemical platform based on screen-printed electrodes on a flexible polyester film for the direct determination of nitrite in the supernatants of lipopolysaccharide (LPS)-stimulated macrophages. The device achieves micromolar sensitivity, with a limit of detection of approximately 1.7 μM, and its results correlate closely with the Griess method (Pearson's r = 0.980, p < 0.001) without any electrode modification. Beyond detection, the sensor was successfully employed to monitor the anti-inflammatory response to three mechanistically distinct drugs (erucin, naproxen, and deflazacort), demonstrating its suitability for probing cellular toxicity in different systems. Combining portability, low cost, and operational simplicity, this platform provides a practical alternative to conventional assays for quickly evaluating the therapeutic efficacy in in-vitro models.

Duchenne muscular dystrophy (DMD) is an X-linked neuromuscular disorder characterized by oxidative stress, chronic inflammation, and skeletal muscle degeneration. Although deflazacort is the standard treatment, its long-term use is associated with significant adverse effects. Since impaired hydrogen sulfide (H2S) signaling contributes to DMD pathogenesis, we designed five novel deflazacort−H2S donor hybrids (I−V) through a molecular hybridization strategy combining anti-inflammatory efficacy with controlled sulfide release. All hybrids showed chemical stability under physiological and acidic conditions, while serum stability varied according to the linker and donor structure. Compound IV exhibited the longest half-life and a slow, l-cysteine-dependent H2S release profile. In LPS + IFN-γ-stimulated C2C12 myotubes, compound IV reduced (i) NOx production, (ii) expression of pro-inflammatory and redox-related genes, and (iii) intracellular ROS levels. In vivo, compound IV, at half of the deflazacort dose, restored locomotor performance, providing superior protection against oxidative stress, highlighting its potential as a dose-sparing therapeutic candidate for DMD.