Investigation of Binding‐Site Homology between Mushroom and Bacterial Tyrosinases by Using Aurones as Effectors

Tyrosinase is a copper‐containing enzyme found in plants and bacteria, as well as in humans, where it is involved in the biosynthesis of melanin‐type pigments. Tyrosinase inhibitors have attracted remarkable research interest as whitening agents in cosmetology, antibrowning agents in food chemistry, and as therapeutics. In this context, commercially available tyrosinase from mushroom (TyM) is frequently used for the identification of inhibitors. This and bacterial tyrosinase (TyB) have been the subjects of intense biochemical and structural studies, including X‐ray diffraction analysis, and this has led to the identification of structural homology and divergence among enzymes from different sources. To better understand the behavior of potential inhibitors of TyM and TyB, we selected the aurone family—previously identified as potential inhibitors of melanin biosynthesis in human melanocytes. In this study, a series of 24 aurones with different hydroxylation patterns at the A‐ and B‐rings were evaluated on TyM and TyB. The results show that, depending on the hydroxylation pattern of A‐ and B‐rings, aurones can behave as inhibitors, substrates, and activators of both enzymes. Computational analysis was performed to identify residues surrounding the aurones in the active sites of both enzymes and to rationalize the interactions. Our results highlight similarities and divergence in the behavior of TyM and TyB toward the same set of molecules.     

Partial Characterization of Nine Bacteriocins Produced by Lactic Acid Bacteria Isolated from Cold-Smoked Salmon with Activity against Listeria monocytogenes

Nine LAB bacteriocin-producers, isolated from vacuum-packaged cold-smoked salmon (CSS), were phenotypically and genotypically identified as Lactobacillus curvatus, Lactobacillus delbrueckii, Lactobacillus fermentum, Enterococcus faecium, and Pediococcus acidilactici. Their bacteriocins were partially characterized. The antimicrobial spectrum was determined against Listeria monocytogenes, E. faecalis, E. faecium, and Staphylococcus aureus. The molecular size of bacteriocins ranged from 2.8 to 4.5 kDa. They were inactivated by treatment with proteolytic enzymes but not by lipolytic or glycolytic enzymes. Maximal activity against L. monocytogenes ranged between 800 and 10000 AU/mL at pH 6.5. Most of the bacteriocins maintained full activity in a pH range of 2.0 to 8.0 but were partially or completely inactivated at pH 10.0. After heating at 60°C and 100°C, only two bacteriocins from Lb. curvatus strains partially lost activity. All bacteriocins showed a narrow spectrum of activity and a high anti-listerial activity, which is characteristic of the class IIa bacteriocins. Isolated bacteriocin-producing LAB could be used successfully in the bio-preservation of CSS and development of new potential bio-preservatives for CSS active against L. monocytogenes.     

Synthesis, Characterization and Transesterification Activity of Cross-Linked Styrenic Resins Containing the Triphenyltincarboxylate Moiety Spaced by a Dimethylene from the Aromatic Ring

An organotin monomer, triphenyltin 3-(4-styryl)-propionate (TPTSPr) has been synthesized and copolymerized in different ratios with styrene and 1,4-divinylbenzene in order to obtain resins with catalytic activity in transesterification reactions. The resins and a low molecular weight model compound, triphenyltin 3-(4-ethylphenyl)-propionate (TPT-C2-Pr), mimicking the catalytic co-unit, have been characterized by FT-IR and NMR spectroscopy, with particular attention paid to the coordination at tin and how it correlates to the catalytic activity. The activity of both the resins and of the model compound have been tested in a transesterification model reaction between ethyl acetate and primary alcohol. All the resins show catalytic activity that decreases with increasing content of the active co-unit in the resins, owing to the interaction of the active sites among themselves.     

Biomimetic poly(glycerol sebacate) (PGS) membranes for cardiac patch application

In this study biomimetic poly(glycerol sebacate) PGS matrix was developed for cardiac patch application. The rationale was that such matrices would provide conducive environment for the seeded cells at the interphase with PGS. From the microstructural standpoint, PGS was fabricated into dense films and porous PGS scaffolds. From the biological aspect, biomimetic PGS membranes were developed via covalently binding peptides Tyr-Ile-Gly-Ser-Arg (YIGSR) and Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), corresponding to the epitope sequences of laminin and fibronectin, respectively onto the surface. To improve and enhance homogenous binding of peptides onto the PGS surface, chemical modification of its surface was carried out. A sequential regime of alkaline hydrolysis with 0.01 M NaOH for 5 min and acidification with 0.01 M HCl for 25 s was optimal. More COOH chemical group was exposed without causing deleterious effect on the bulk properties of the polymer as revealed by the physicochemical analysis carried out. HPLC analysis, chemical imaging and ToF-SIMS were able to establish the successful homogenous functionalization of PGS membranes with the peptides. Finally, the developed biomimetic membranes supported the adhesion and growth of rat and human cardiac progenitor cells.     

Studies on the catalytic dechlorination and abatement of chlorided VOC: the cases of 2-chloropropane, 1,2-dichloropropane and trichloroethylene

The conversion of monochloropropanes and dichloropropanes over acid catalysts has been investigated in the presence of oxygen. In the temperature range of 450–550 K, dehydrochlorination of monochloropropanes to propene and HCl occurs selectively over silica–alumina, while significant formation of chlorinated by-products is observed over ZSM5 zeolite catalyst even at higher temperatures. Dichloropropanes conversion over silica–alumina catalyst gives rise mainly to chloropropenes in the temperature range 500–700 K. CO_x are predominant products only at the highest reaction temperatures (just above 700 K). Water vapor in the feed only slightly affects conversions and selectivities. Deactivation processes occur upon dichloropropane conversion, mainly due to coke deposition.

The conversion of highly chlorinated compounds, such as trichloroethylene (TCE) has been tested over silica–alumina and over HY zeolite in the presence of water vapor in the so-called “steam reforming” conditions (HVOC:water=1:2). With diluted feed (1200 ppm) on HY, reaction occurs above 800 K and formation of chlorinated by-products is minimized, CO_x being the main reaction products. At higher HVOC concentrations conversion is obtained at even lower temperature (600 K), but no more negligible by-products formation has been detected. In our conditions zeolite catalyst is more effective in TCE total conversion than silica–alumina.     

Isostructural organic–inorganic hybrids of P,P-diphenyl-methylenediphosphinate (CH2(P(Ph)O2)2) with divalent transition metals

New organic–inorganic hybrid materials have been synthesized by reaction in water solution of manganese, cobalt or nickel acetates with P,P^′-diphenylmethylenediphosphinic acid and all the two-dimensional structure coordination polymers obtained have been found to be isomorphous.     

Volatile compounds from Chétoui olive oil and variations induced by growing area

Fruits from the same variety of Olea europaea L., grown under different environmental conditions in the north of Tunisia, were harvested at the same ripening degree and immediately processed. The volatile profile of virgin olive oils was established using solid phase, micro-extraction (SPME) and gas chromatography-mass spectrometry (GC-MS). Compounds belonging mainly to the following chemical classes characterised the volatile profiles: esters, aldehydes, ketons, aliphatic alcohols and hydrocarbons. Significant differences in the proportions of volatile constituents from oils of different geographical origins were detected and the major volatile in approximately 50% of the oil samples was the aldehyde (E)-2-hexenal. The results suggest that, beside the genetic factor, environmental conditions influence the volatile formation.     

Cell and matrix morpho-functional analysis in chondrocyte micromasses

Micromass cultures represent a convenient means of studying chondrocyte physiology in the context of a tridimensional culture model. In this study, we present the first ultrastructural analysis of the distribution and organization of the extracellular components in micromasses in comparison with their cartilaginous counterparts. Primary chondrocytes obtained from osteoarthritis patients were pelleted in micromasses. Transmission electron microscopy and immunofluorescence were used to evaluate the distribution of major extracellular matrix proteins, i.e., aggrecan, chondroitin-4-sulfate, chondroitin-6-sulfate, and collagen I and II. Both approaches revealed a number of morphological features shared by micromass and cartilage chondrocytes. In particular, in micromasses, chondrocytes are in close contact with an organized extracellular matrix that adequately mimics that of cartilage. Cells were observed to establish specialized junctions for cell-extracellular matrix crosstalk. Noteworthy, cells seem endowed in a chondroitin sulfate-rich microenvironment, and thus possibly ensuring the immobilization of chemokines, a family of molecules emerging in osteoarthritis pathogenesis, in a haptotactic-like gradient to the chondrocytes, which facilitates the binding to their receptors. To determine the suitability of this model to investigate osteoarthritis pathogenesis, a potential apoptotic stimulus (endothelial IL-8) was used, and ultrastructural analysis assessed apoptosis induction. Micromass cultures were proved to be an experimental technique providing a large number of properly differentiated chondrocytes, and thus allowing reliable biochemical and morphological studies. They represent, therefore, a novel approach to osteoarthritis investigation that promises more thorough understanding of chondrocyte physiology in osteoarthritis.     

Mitochondrial alterations and autofluorescent conversion of Candida albicans induced by histatins

The mechanism of the candidacidal activity of histatins 3 and 5 (Hst) is still a matter of debate. Previous studies have indicated that Hst induce cell permeabilization, generation of reactive oxygen species (ROS) by mitochondria, inhibition of the respiratory chain, and energy-dependent cytotoxic release of ATP. On the other hand, the multiplicity of effects and the apparent contrast between experimental data continue to render the mechanism of Hst-induced killing of C. albicans unclear. In this investigation, using fluorescent probes (the potential-sensitive mitochondrial probe tetramethylrhodamine methyl ester perchlorate, TMRM; the ROS-sensitive probe dihydrofluorescein diacetate, DHF; the membrane-impermeant probe, calcein) and autofluorescence data we observed that Hst induce ROS generation by mitochondria undergoing a high energy swelling condition, accompanied by oxidation of cytosolic NAD(P)H and mitochondrial flavoproteins. ROS generation and swelling, attributable to an inhibition of the respiratory chain and to impairment of the K/H-exchanger, were followed by mitochondrial depolarization. Mitochondrial changes were accompanied by massive calcein influx, indicative of cell permeabilization, and prominent alterations of the cell size, shape, and optical density. The loss of proliferative activity was correlated, on a single cell basis, to the acquisition of a lipofuscin-like autofluorescence.     

Thermo‐oxidative stabilization of poly(lactic acid)‐based nanocomposites through the incorporation of clay with in‐built antioxidant activity

In this work, an innovative approach to overcome the issue of the poor thermo‐oxidative stability of polymer/clay nanocomposites is proposed. Specifically, biodegradable poly(lactic acid) (PLA)‐based nanocomposites, containing organo‐modified clay with in‐built antioxidant activity, were prepared. Through a two‐step chemical protocol, a hindered phenol antioxidant was chemically linked to the ammonium quaternary salt which was then intercalated between the clay platelets [(AO)OM‐Mt]. The nanocomposites were characterized and their thermo‐oxidative stability during melt processing and under long‐term thermal test conditions was investigated. PLA nanocomposites containing the (AO)OM‐Mt showed higher oxidative stability, along with better clay dispersion, compared to PLA‐nanocomposites containing commercial clay and a free hindered phenol antioxidant. Obtained results can be explained considering that (AO)OM‐Mt may act locally, at the interface, between the silicate layers and the polymer macromolecules, thus contributing to the observed improved stability of the polymer both during processing and under long‐term thermal‐oxidative conditions. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44974.