Cigarette smoke (CS) is a risk factor for inflammatory diseases, such as atherosclerosis. CS condensate (CSC) contains lipophilic components that may represent a systemic cardiac risk factor. To better understand CSC effects, we incubated mouse and human aortic smooth muscle cells (SMCs) with CSC. We evaluated specific markers for contractile [i.e., actin, aortic smooth muscle (ACTA2), calponin-1 (CNN1), the Kruppel-like factor 4 (KLF4), and myocardin (MYOCD) genes] and inflammatory [i.e., IL-1β, and IL-6, IL-8, and galectin-3 (LGALS-3) genes] phenotypes. CSC increased the expression of inflammatory markers and reduced the contractile ones in both cell types, with KLF4 modulating the SMC phenotypic switch. Next, we performed a mass spectrometry-based differential proteomic approach on human SMCs and could show 11 proteins were significantly affected by exposition to CSC (FC ≥ 2.7, p ≤ 0.05). These proteins are active in signaling pathways related to expression of pro-inflammatory cytokines and IFN, inflammasome assembly and activation, cytoskeleton regulation and SMC contraction, mitochondrial integrity and cellular response to oxidative stress, proteostasis control via ubiquitination, and cell proliferation and epithelial-to-mesenchymal transition. Through specific bioinformatics resources, we showed their tight functional correlation in a close interaction niche mainly orchestrated by the interferon-induced double-stranded RNA-activated protein kinase (alternative name: protein kinase RNA-activated; PKR) (EIF2AK2/PKR). Finally, by combining gene expression and protein abundance data we obtained a hybrid network showing reciprocal integration of the CSC-deregulated factors and indicating KLF4 and PKR as the most relevant factors.

Epiretinal membranes (ERMs) are sheets of tissue that pathologically develop in the vitreoretinal interface leading to progressive vision loss. They are formed by different cell types and by an exuberant deposition of extracellular matrix proteins. Recently, we reviewed ERMs’ extracellular matrix components to better understand molecular dysfunctions that trigger and fuel the onset and development of this disease. The bioinformatics approach we applied delineated a comprehensive overview on this fibrocellular tissue and on critical proteins that could really impact ERM physiopathology. Our interactomic analysis proposed the hyaluronic-acid-receptor cluster of differentiation 44 (CD44) as a central regulator of ERM aberrant dynamics and progression. Interestingly, the interaction between CD44 and podoplanin (PDPN) was shown to promote directional migration in epithelial cells. PDPN is a glycoprotein overexpressed in various cancers and a growing body of evidence indicates its relevant function in several fibrotic and inflammatory pathologies. The binding of PDPN to partner proteins and/or its ligand results in the modulation of signaling pathways regulating proliferation, contractility, migration, epithelial–mesenchymal transition, and extracellular matrix remodeling, all processes that are vital in ERM formation. In this context, the understanding of the PDPN role can help to modulate signaling during fibrosis, hence opening a new line of therapy.

The production of food with a naturally enriched protein content is a strategic response to the growing global demand for sustainable protein sources. Wood distillate (WD), a by-product of the pyrolysis of woody biomass, has previously been shown to increase the protein concentration and bioavailability in chickpea seeds. Here, we evaluated the effect of 0.5% (v/v) WD soil drenching on chickpea productivity, nutritional profile, and proteomic pattern. WD treatment significantly improved the yield by increasing plant biomass (+144%), number of pods and seeds (+148% and +147%), and seed size (diameter: +6%; weight: +25%). Nutritional analyses revealed elevated levels of soluble proteins (+15%), starch (+11%), fructose (+135%), and polyphenols (+14%) and a greater antioxidant capacity (25%), alongside a reduction in glucose content, albeit not statistically significant, suggesting an unchanged or even lowered glycemic index. Although their concentration decreased, Ca (−31%), K (−12%), P (−5%), and Zn (−14%) in WD-treated plants remained within normal ranges. To preliminary assess the quality and safety of the protein enrichment, a differential proteomic analysis was performed on coarse flours from individual seeds. Despite the higher protein content, the overall protein profiles of the WD-treated seeds showed limited variation, with only a few storage proteins, identified as legumin and vicilin-like isoforms, being differentially abundant. These findings indicate a general protein concentration increase without a major alteration in the proteoform composition or differential protein synthesis. Overall, WD emerged as a promising and sustainable biostimulant for chickpea cultivation, capable of enhancing both yield and nutritional value, while maintaining the proteomic integrity and, bona fide, food safety.