Probing interfacial organization in surface monolayers using tethered pyrene. 1. Structural mediation of electron and proton access to adsorbates

We have synthesized and characterized a family of self-assembled monolayers containing pyrene derivatives on gold and indium-doped tin oxide (ITO) substrates. The covalently bound pyrene functionalities serve as either spectroscopic or electrochemical probes of their immediate environment, and we explore their electrochemical response in this paper. When these compounds are the only constituents bound to the interfaces, the molecules enjoy significant structural freedom. The addition of aliphatic adsorbates to the interfaces serves to place the pyrene derivatives in a more restricted environment. Cyclic voltammetry shows that the organization of a monolayer with pyrene derivatives, and the position of the terminal pyrene within such monolayer, depend sensitively on the length of the pyrene tether and the presence or absence of aliphatic interfacial species, as well as the identity of the substrate.     

In vitro evaluation of potential complexation between bovine insulin and bovine serum albumin

The objective of this study was to examine the possible binding of bovine insulin (BI) with bovine serum albumin (BSA) to form a new potential diabetogenic irreversible complex protein. Several preparations of BSA and BI were prepared. Both capillary electrophoresis and spectrophotometric analysis were undertaken to test the possibility of complexation between BI and BSA. HPLC was used to test whether the potential complex of BI and BSA is reversible or irreversible. The optimum deviation between the real and calculated absorbances was observed at a BI/BSA ratio of 2. Moreover, the migration time of BI decreased substantially with increasing ratio of BI to BSA until it became almost constant at equal molar ratio of BI/BSA. While the majority of the 2:1 BI–BSA sample detached during the HPLC analysis, which confirms the reversible character of BI–BSA binding, the HPLC chromatogram also emphasizes the formation of an irreversible complexation between the two proteins. This study provides evidence of the formation of reversible and irreversible new BI–BSA complexes under physiological conditions. This highlights the importance of examining the possible diabetogenicity of BI–BSA complex in genetically susceptible people. Copyright © 2013 John Wiley & Sons, Ltd.     

Probing interfacial organization in surface monolayers using tethered pyrene. 2. Spectroscopy and motional freedom of the adsorbates

We have studied the steady-state and time-resolved emission spectroscopy of the pyrene-containing monolayers reported in the previous article, where in this work we have bound the monolayers to SiO(x). We find that these monolayer structures are sensitive to the identity of the solvent overlayer, with the solvent playing a significant role in the organization of the surface-bound monolayers. We discuss our findings in the context of the known polarity dependence of the pyrene emission spectrum and find that the motional freedom of the chromophores varies with both the monolayer composition and the identity of the solvent overlayer. Our data point to the importance of neighbor-neighbor interactions within the monolayer structures in mediating the motional freedom of the tethered pyrene chromophores.     

The Importance of Nutraceuticals in COVID-19: What’s the Role of Resveratrol?

Since COVID-19 has affected global public health, there has been an urgency to find a solution to limit both the number of infections, and the aggressiveness of the disease once infected. The main characteristic of this infection is represented by a strong alteration of the immune system which, day by day, increases the risk of mortality, and can lead to a multiorgan dysfunction. Because nutritional profile can influence patient’s immunity, we focus our interest on resveratrol, a polyphenolic compound known for its immunomodulating and anti-inflammatory properties. We reviewed all the information concerning the different roles of resveratrol in COVID-19 pathophysiology using PubMed and Scopus as the main databases. Interestingly, we find out that resveratrol may exert its role through different mechanisms. In fact, it has antiviral activity inhibiting virus entrance in cells and viral replication. Resveratrol also improves autophagy and decreases pro-inflammatory agents expression acting as an anti-inflammatory agent. It regulates immune cell response and pro-inflammatory cytokines and prevents the onset of thrombotic events that usually occur in COVID-19 patients. Since resveratrol acts through different mechanisms, the effect could be enhanced, making a totally natural agent particularly effective as an adjuvant in anti COVID-19 therapy.     

PHOTOINACTIVATION (365 NM) OF VACCINIA AND HERPES SIMPLEX VIRUSES INDUCED BY A NEW BUILT-IN DNA PHOTOSENSITIZER: 4-THIOTHYMIDINE

The thymidine analogue 4-thiothymidine (s⁴T) strongly absorbs light at wavelengths in the UVA range (Λ_max 335 nm) and we have examined the photoinactivation of vaccinia and herpes simplex viruses grown in the presence of this nucleoside. The cells used in this study (Vero, mouse 1D-TK⁺) were able to grow at the same rate when cultured in the presence of 2 mM s⁴T or 2 mM thymidine, albeit at a slower rate than control cells. Consistent with this finding, viruses grown in the presence of1–4 mM s⁴T were obtained in reduced yield but retained full infectivity. Both viruses were specifically inactivated by irradiation with 365 nm light and their photosensitivity, as measured by the initial slope of the inactivation curve, increased in parallel with the concentration of s⁴T added to the culture medium. More than 90% of vaccinia virus grown in the presence of 4 mM s⁴T was inactivated. Organomercurial agarose chromatography of sheared DNA isolated from vaccinia virus grown in the presence of 2 mM s⁴T showed that approximately 2.5% of DNA fragments were specifically retained, as compared to 0.2% for control DNA. This value corresponds to at least one s4T residue incorporated per 30 000 nucleotides of vaccinia virus DNA. In fact, it is likely that this ratio is actually approximately 10 times higher because of the incomplete retention of control thiolated oligodeoxynucleotides. The incorporation of s⁴T into vaccinia virus DNA was required for photoinactivation as (1) the expression of a viral or cellular thymidine kinase was required to confer photosensitivity, and (2) virus plaque reduction assays revealed that maximal photosensitivity coincided with the first rounds of viral DNA replication. The photo-inactivated virus was unable to induce detectable synthesis of several early proteins after infection of cells. These data show that s⁴T is incorporated into the DNA of vaccinia virus grown in the presence of the analogue and then behaves as a built-in UVA light photosensitizer.     

Interrogating interfacial organization in planar bilayer structures

The field of biomimetic planar lipid membranes is finding increased importance as the need to devise sensing systems for biologically important species increases. We approach this area with an eye toward understanding how to interrogate local organization in these complex media. Our primary tools for this purpose are spectroscopy and electrochemistry, where imbedded reporter molecules serve as the information transducers. We use Langmuir-Blodgett (LB) and Langmuir-Schaefer (LS) methods to construct planar lipid membranes on hydrophilic solid substrates (Au for electrochemistry, SiO x for spectroscopy). Pyrene tethered to the substrate acts as our probe and AC voltammetry was used to evaluate its redox kinetics, showing slow, distance independent electron transfer between the pyrene moieties and the electrode for both monolayer and bilayer systems. Time-resolved fluorescence data indicate that tethered pyrene resides in a highly rigid environment and that the addition of the top lipid leaflet improves the organization of the bottom lipid leaflet. These data point to the cooperative effect of the bilayer leaflets in creating a system that is comparatively rigid on short length scales, and capable of mediating motion and accessibility of imbedded species.     

Strong inclusion of inorganic anions into β-cyclodextrin immobilized to gold electrode

The inclusion of inorganic anions such as SO(4)(2-), NO(3)(-), and HPO(4)(2-) into the cavity of β-cyclodextrin monolayers on Au was examined by X-ray photoelectron spectroscopy (XPS), a quartz crystal microbalance (QCM), and chronocoulometric measurements of the competitive inclusion with ferrocene. The inclusion amounts of ferrocence in 0.2 M Na(2)SO(4), NaNO(3), and Na(2)HPO(4) solutions were less than 6% of the adsorption amount of β-cyclodextrin on Au, resulting in the apparent inhibition of the ferrocene redox reaction. The surface association constants of these anions reached about 10 on a logarithmic scale and were much higher than those for the inclusion of common organic guest compounds. A stronger anion inclusion was also demonstrated by the QCM response corresponding to the replacement of a preincluded organic guest with sulfate upon the injection of the sulfate solution. Quantitative analysis of the XPS data suggested a 1:1 association for each of these anions per surface β-cyclodextrin. There was no detectable inclusion for ClO(4)(-), Cl(-), and Br(-).