Synthesis, Molecular Characterization and Preliminary Antioxidant Activity Evaluation of Quercetin Fatty Esters

Quercetin shows interesting pharmacological effects, but its use in topical applications is limited by its low skin permeability and solubility. In this work, the synthesis of highly lipophilic quercetin esters with oleic, linoleic and linolenic acid useful as topical quercetin prodrugs is reported. Partial OH esterification is advisable to maintain the antioxidant activity of these compounds; tetraesters and triesters can be achieved by modulating the reaction conditions utilized for the total esterification of quercetin. The chemical structures of the esters were proven by spectroscopic techniques; quantum chemical NMR calculation were mandatory to unequivocally assign the free position in triesters. Finally, the antioxidant activity of all the synthesized compounds was determined by the 2,2-diphenyl-1-picryl-hydrazyl method and by 2,2-azinobis(3-ethyl-benzothiazoline-6-sulfonic acid) assay.     

Peptide-Lipid Interactions: Experiments and Applications

The interactions between peptides and lipids are of fundamental importance in the functioning of numerous membrane-mediated cellular processes including antimicrobial peptide action, hormone-receptor interactions, drug bioavailability across the blood-brain barrier and viral fusion processes. Moreover, a major goal of modern biotechnology is obtaining new potent pharmaceutical agents whose biological action is dependent on the binding of peptides to lipid-bilayers. Several issues need to be addressed such as secondary structure, orientation, oligomerization and localization inside the membrane. At the same time, the structural effects which the peptides cause on the lipid bilayer are important for the interactions and need to be elucidated. The structural characterization of membrane active peptides in membranes is a harsh experimental challenge. It is in fact accepted that no single experimental technique can give a complete structural picture of the interaction, but rather a combination of different techniques is necessary.     

Assessment of production efficiency,physicochemical properties and storage stability of spray-dried propolis,a natural food additive,using gum Arabic and OSA starch-based carrier systems

The aim of this work was to obtain propolis in a powder, alcohol-free, water-dispersed and shelf-stable form. Propolis extract was spray-dried using gum Arabic and octenyl succinic anhydride (OSA) starch as carriers in two different weight ratios (1:4 and 1:6). Spray-dried propolis samples were evaluated for morphology, moisture, water activity, water dispersibility, hygroscopicity, particle size, particle distribution, entrapping efficiency, stability, isotherms and antioxidant properties. The spray-drying process produced round particles with sizes ranging from 15 to 24 μm. This process preserved the antioxidant activity of propolis and also allowed propolis to be obtained in a powder form, which was stable during storage at room temperature, had low hygroscopicity and was highly dispersible in cold water. The application of this technology could increase the use of propolis in various industrial applications, such as an antimicrobial and as an antioxidant in food.     

The influence of drying air temperature on the physical properties of dried and rehydrated eggplant

Drying processes generally cause volume and surface change of foodstuffs. Information on the porous structure and the mechanical properties of dried food products is needed for determining food quality, process design and estimating properties such as density and moisture diffusivity.In this work we investigated the structural changes induced in eggplant by convective air drying at four different temperatures (40, 50, 60 and 70 °C) and their effect on the subsequent rehydration process. Drying and rehydration kinetic curves were also measured.The changes in physical properties, such as porosity, pore-size distribution and bulk density were determined by Hg porosimetry, scanning electron microscopy and optical microscopy while their effect on the textural characteristics by dynamometric measurements.As expected, the increase of the drying air temperature causes shorter drying times. The drying temperature influences strongly the microstructure of dried samples: the porosity increases with the air temperature, but the structure is better preserved at intermediate temperature (60 °C) as confirmed by the lower firmness values with respect to the other dehydrated samples (40, 50 and 70 °C). In these latter, the longer drying time and the higher temperature, respectively, causes the development of a wrinkled structure. In particular, at 70 °C the structure of dehydrated samples appears totally broken with a consequent faster water uptake during rehydration.     

Senescent CD4+CD28− T Lymphocytes as a Potential Driver of Th17/Treg Imbalance and Alveolar Bone Resorption during Periodontitis

Senescent cells express a senescence-associated secretory phenotype (SASP) with a pro-inflammatory bias, which contributes to the chronicity of inflammation. During chronic inflammatory diseases, infiltrating CD4⁺ T lymphocytes can undergo cellular senescence and arrest the surface expression of CD28, have a response biased towards T-helper type-17 (Th17) of immunity, and show a remarkable ability to induce osteoclastogenesis. As a cellular counterpart, T regulatory lymphocytes (Tregs) can also undergo cellular senescence, and CD28⁻ Tregs are able to express an SASP secretome, thus severely altering their immunosuppressive capacities. During periodontitis, the persistent microbial challenge and chronic inflammation favor the induction of cellular senescence. Therefore, senescence of Th17 and Treg lymphocytes could contribute to Th17/Treg imbalance and favor the tooth-supporting alveolar bone loss characteristic of the disease. In the present review, we describe the concept of cellular senescence; particularly, the one produced during chronic inflammation and persistent microbial antigen challenge. In addition, we detail the different markers used to identify senescent cells, proposing those specific to senescent T lymphocytes that can be used for periodontal research purposes. Finally, we discuss the existing literature that allows us to suggest the potential pathogenic role of senescent CD4⁺CD28⁻ T lymphocytes in periodontitis.     

Rationally designed inhibitors as tools for comparing the mechanism of squalene-hopene cyclase with oxidosqualene cyclase

The inhibition of squalene-hopene cyclase (SHC) (E.C. 5.4.99.-), an enzyme of bacterial membranes catalyzing the formation of pentacyclic sterol-like triterpenes, was studied by using different classes of compounds originally developed as inhibitors of oxidosqualene cyclase (OCS) (E.C. 5.4.99.7), the enzyme of eukaryotes responsible for the formation of tetracyclic precursors of sterols. The mechanism of cyclization of squalene by SHC, beginning with a protonation of the 2,3 double bond by an acidic residue of the enzyme, followed by a series of electrophilic additions of the carbocationic intermediates to the double bonds, is similar to the mechanism of cyclization of 2,3-oxidosqualene by OSC. The inhibitors studied included: (i) analogs of the carbocationic intermediates formed during cyclization, such as aza-analogs of squalene and 2,3-oxidosqualene; (ii) affinity-labeling inhibitors bearing a methylidene reactive group; and (iii) vinyldioxidosqualenes and vinylsulfide derivatives of the substrates. Comparison of the results obtained with the two enzymes, SHC and OSC, showed that many of the most effective inhibitors of OSC were also able to inhibit SHC, while some derivatives acted as specific inhibitors. Differences could be easily explained on the basis of the different substrate specificity of the two enzymes.     

Antibody library selection by the {beta}-lactamase protein fragment complementation assay

Protein fragment complementation assay (PCA) is based on the interaction between two protein partners (e.g. target antigen and antibody), which are genetically fused to the two halves of a dissected marker protein. Binding of the two partners reassembles the marker protein and hence reconstitutes its activity. In this work we have developed the first application of beta-lactamase-based PCA for the isolation of single chain Fv fragments (scFvs) binding to the human receptor RON from a na?ve library. Specific scFvs with the ability to immunoprecipitate could be isolated after a single round of PCA selection from an scFv repertoire previously pre-selected by phage display. Furthermore, the PCA was used to successfully map the epitopes recognized by the selected scFvs by screening them against a small library of random RON fragments.     

Engineering a pharmacologically superior form of leptin for the treatment of obesity

Leptin plays a central role in the homeostasis of body weight through its regulatory effects on appetite and energy expenditure, yet in trials as a therapeutic agent for the treatment of obesity in humans it has been disappointing. The poor clinical efficacy of leptin results from its short circulating half-life, low potency and poor solubility, necessitating large and frequent doses to obtain even modest clinical benefit. Engineered Fc-leptin immunofusins, consisting of the Fc fragment of an immunoglobulin gamma chain followed by leptin, exhibit improved pharmacological properties with very consistent and potent biological activities. Furthermore, in extending the circulating half-life of the protein in vivo from a few minutes for leptin to many hours for Fc-leptin, these proteins have the potential to reduce drastically the dosage and frequency of administration required to obtain clinical benefit. The results of this study show that the engineered leptin immunofusins described here have significantly enhanced pharmacological properties in comparison with the recombinant leptin that was used in clinical trials. As such, they could represent an important step towards a therapeutically superior form of leptin if the disappointing performance of leptin in early clinical trials was due to its poor pharmacological properties rather than any conceptual weakness in the strategy of using leptin for the treatment of obesity and its related disorders.     

UV‐Cured Polysiloxane Epoxy Coatings Containing Titanium Dioxide as Photosensitive Semiconductor

UV‐cured polysiloxane epoxy coatings containing titanium dioxide were prepared by means of a cationic photopolymerization process. A good distribution of the inorganic filler was achieved within the polymeric network with an average size dimension of around 500 nm. UV‐vis analysis performed on organic dye (methylene blue) stained coatings showed a high efficiency of the titania photocatalytic activity: a complete degradation of the dye on the coating surface is reached after 60 min of UV irradiation without affecting the matrix photo‐degradation.