Proprotein Convertase Subtilisin Kexin Type 9 Inhibitors Reduce Platelet Activation Modulating ox-LDL Pathways

Background: Proprotein convertase subtilisin kexin type 9 inhibitors (PCSK9i) lower LDL-cholesterol and slow atherosclerosis preventing cardiovascular events. While it is known that circulating PCSK9 enhances platelet activation (PA) and that PCSK9i reduce it, the underlying mechanism is not still clarified. Methods: In a multicenter before–after study in 80 heterozygous familial hypercholesterolemia (HeFH) patients on treatment with maximum tolerated statin dose ± ezetimibe, PA, soluble-NOX2-derived peptide (sNOX2-dp), and oxidized-LDL (ox-LDL) were measured before and after six months of PCSK9i treatment. In vitro study investigates the effects of plasma from HeFH patients before and after PCK9i on PA in washed platelets (wPLTs) from healthy subjects. Results: Compared to baseline, PCSK9i reduced the serum levels of LDL-c, ox-LDL, Thromboxane (Tx) B2, sNOX2-dp, and PCSK9 (p < 0.001). The decrease of TxB2 correlates with that of ox-LDL, while ox-LDL reduction correlated with PCSK9 and sNOX2-dp delta. In vitro study demonstrated that wPLTs resuspended in plasma from HeFH after PCSK9i treatment induced lower PA and sNOX2-dp release than those obtained using plasma before PCSK9i treatment. This reduction was vanished by adding ox-LDL. ox-LDL-induced PA was blunted by CD36, LOX1, and NOX2 inhibition. Conclusions: PCSK9i treatment reduces PA modulating NOX2 activity and in turn ox-LDL formation in HeFH patients.     

Diphenyl-Methane Based Thyromimetic Inhibitors for Transthyretin Amyloidosis

Thyromimetics, whose physicochemical characteristics are analog to thyroid hormones (THs) and their derivatives, are promising candidates as novel therapeutics for neurodegenerative and metabolic pathologies. In particular, sobetirome (GC-1), one of the initial halogen-free thyromimetics, and newly synthesized IS25 and TG68, with optimized ADME-Tox profile, have recently attracted attention owing to their superior therapeutic benefits, selectivity, and enhanced permeability. Here, we further explored the functional capabilities of these thyromimetics to inhibit transthyretin (TTR) amyloidosis. TTR is a homotetrameric transporter protein for THs, yet it is also responsible for severe amyloid fibril formation, which is facilitated by tetramer dissociation into non-native monomers. By combining nuclear magnetic resonance (NMR) spectroscopy, computational simulation, and biochemical assays, we found that GC-1 and newly designed diphenyl-methane-based thyromimetics, namely IS25 and TG68, are TTR stabilizers and efficient suppressors of TTR aggregation. Based on these observations, we propose the novel potential of thyromimetics as a multi-functional therapeutic molecule for TTR-related pathologies, including neurodegenerative diseases.     

Chronically Elevated Exogenous Glucose Elicits Antipodal Effects on the Proteome Signature of Differentiating Human iPSC-Derived Pancreatic Progenitors

The past decade revealed that cell identity changes, such as dedifferentiation or transdifferentiation, accompany the insulin-producing β-cell decay in most diabetes conditions. Mapping and controlling the mechanisms governing these processes is, thus, extremely valuable for managing the disease progression. Extracellular glucose is known to influence cell identity by impacting the redox balance. Here, we use global proteomics and pathway analysis to map the response of differentiating human pancreatic progenitors to chronically increased in vitro glucose levels. We show that exogenous high glucose levels impact different protein subsets in a concentration-dependent manner. In contrast, regardless of concentration, glucose elicits an antipodal effect on the proteome landscape, inducing both beneficial and detrimental changes in regard to achieving the desired islet cell fingerprint. Furthermore, we identified that only a subgroup of these effects and pathways are regulated by changes in redox balance. Our study highlights a complex effect of exogenous glucose on differentiating pancreas progenitors characterized by a distinct proteome signature.     

Anti-listerial activity of a polymeric film coated with hybrid coatings doped with Enterocin 416K1 for use as bioactive food packaging

In this study, Enterocin 416K1, a bacteriocin produced by Enterococcus casseliflavus IM 416K1, was entrapped in an organic-inorganic hybrid coating applied to a LDPE (low-density polyethylene) film for its potential use in the active food packaging field. The antibacterial activity of the coated film was evaluated against Listeria monocytogenes NCTC 10888 by qualitative modified agar diffusion assay, quantitative determination in listeria saline solution suspension and direct contact with artificially contaminated food samples (frankfurters and fresh cheeses) stored at room and refrigeration temperatures. All investigations demonstrated that enterocin-activated coatings have a good anti-listeria activity. Qualitative tests showed a clear zone of inhibition in the indicator lawn in contact with and around the coated film. During the quantitative antibacterial evaluation the L. monocytogenes viable counts decreased to 1.5 log units compared to the control. The inhibitory capability was confirmed also in food-contact assays. In all food samples packed with coated films we observed a significant decrease in L. monocytogenes viable counts in the first 24 h compared to the control. This difference was generally maintained up to the seventh day and then decreased, with the exception of the cheese samples stored at refrigeration temperature.     

Influence of oxygen on the sputtering of aluminum oxide for the surface passivation of crystalline silicon

While sputtering has been shown to be capable of depositing aluminum oxide suitable for surface passivation, the mechanisms for this are yet to be firmly established and its potential realized. In this paper, we investigate the relationships between the oxygen in the sputtering process to the resulting composition of the deposited film and the surface passivation obtained. We find that surface passivation is not strongly dependent on the bulk composition of the film. Instead the results indicate that the interfacial silicon oxide layer that forms after annealing between the aluminum oxide film and the silicon is a much more important factor; it is this combined structure of aluminum oxide, silicon oxide and silicon that is crucial for obtaining negative charges and excellent surface passivation.     

Supercritical carbon dioxide-treated blood orange juice as a new product in the fresh fruit juice market

The work described here deals with the effectiveness of using high-pressure carbon dioxide treatment (HPCD) to stabilise freshly squeezed blood orange juice. Technical planning of a continuous high-pressure supercritical carbon dioxide pilot system, suitable for development on an industrial scale, was carried out in our lab. To determine the optimal operating conditions (temperature, pressure, and CO₂/juice ratio), three different experimental trials were carried out. The first trial was conducted at 230 bar, 36 ± 1 °C, 5.08 L/h juice flow rate, and 3.91 L/h CO₂ flow rate, corresponding to a g_CO2/g_juice ratio of 0.770. The second trial utilised the same conditions except that the operative pressure was reduced (130 bar). The third trial was carried out at 130 bar, 36 ± 1 °C, 5.08 L/h juice flow rate, 1.96 L/h CO₂ flow rate, corresponding to a 0.385 g_CO2/g_juice ratio. The effects of processing were evaluated by determining physicochemical, antioxidant, and microbiological parameters of the treated juices. In addition, once the best operative parameters had been determined, physicochemical, antioxidant, microbiological and sensory evaluation of fresh blood orange juice stabilised by HPCD treatment was carried out during refrigerated storage of juices at 4 ± 1 °C for thirty days. The results showed that HPCD treatment cannot be considered as an alternative to traditional thermal methods but as a new mild technology for producing a stabilised blood orange juice with a shelf-life of 20 days.Industrial relevanceBlood oranges are the main cultivated varieties of Citrus sinensis (L.) Osbeck in Italy. Freshly squeezed blood orange juice exert a high antiradical and antioxidant activity, due to its rich phenolic profile, but its preservation is usually assured by thermal treatment which affects its nutritional and sensory value. In this study we proposed a “milder” continuous HPCD process suitable for implementation on an industrial scale. The HPCD stabilised juice retains its physicochemical, antioxidant, and sensory properties and could be placed within a new retail framework, namely, that of fresh juices with a shelf-life of 20 days.     

Storage-time effects on olive oil DNA assessed by Amplified Fragments Length Polymorphisms

In this study Amplified Fragments Length Polymorphisms (AFLPs) analysis was applied on DNA extracted from different monovarietal olive oils. The aim was to study how the length of storage after milling of the oil can affect the use of DNA as an analyte for molecular traceability. Results, all assessed by statistical analyses, showed that the authentication of olive oil with molecular methods should be performed within a month from olive oil production. After this period, a significant decrease of quality of DNA extracted from olive oil was observed, with a consequent loss of information, that can affect the reliability of the results.     

Phenotypic and genotypic characterization of Oenococcus oeni strains isolated from Italian wines

A phenotypic and genotypic characterization of 84 Oenococcus oeni isolates from Italian wines of different oenological areas was carried out. Numerical analysis of fatty acid profiles grouped the isolates into two clusters at low level of similarity (63%), the minor cluster containing seven isolates besides the type and the reference strains. Forthy-eight O. oeni isolates, representative of the two clusters, showed no differences in their metabolic properties (heterolactic fermentation pattern, citrate degradation capability and formation of some secondary metabolites). Moreover, the analysis of species-specific randomly amplified polymorphic DNA and 16S-23S rDNA intergenic spacer region polymorphism as well as the sequence-specific separation of V3 region from 16S rDNA by denaturing gradient gel electrophoresis demonstrated a substantial homogeneity among the isolates. On the basis of ApaI Pulsed Field Gel Electrophoresis (PFGE) restriction patterns, the 84 isolates were grouped into five different clusters at 70% similarity, but no correlation with the phenotypic groups could be demonstrated. However, by combining phenotypic and genotypic data, the 84 O. oeni isolates grouped into eight phenotypic-genotypic combined profiles and a relationship between the origin of the isolates and their combined profile became evident, so that a sort of strain specificity can be envisaged for each wine-producing area.