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.     

Stereochemical Outcome of the Biocatalysed Reduction of Activated Tetrasubstituted Olefins by Old Yellow Enzymes 1–3

The ene‐reductase‐mediated reactions of (E)‐ and (Z)‐stereoisomers of tetrasubstituted activated alkenes were performed by means of isolated old yellow enzymes (OYEs) 1–3. The comprehension of the resulting data required a careful analysis of the stereochemical course of this kind of reaction. The investigation of the bioreduction of tetrasubstituted alkenes allowed us to appreciate the contribution of several factors (configuration of the starting alkene, mechanism of hydrogen addition, substrate binding mode) to the overall stereochemistry of the reaction.     

Single electron tunneling in large scale nanojunction arrays with bisferrocene-nanoparticle hybrids

We report on the fabrication and single electron tunneling behaviour of large scale arrays of nanogap electrodes bridged by bisferrocene-gold nanoparticle hybrids (BFc-AuNP). Coulomb staircase was observed in the low temperature current-voltage curves measured on the junctions with asymmetric tunnel barriers. On the other hand, junctions with symmetric tunneling barrier exhibited mere nonlinear current voltage characteristics without discrete staircase. The experimental results agreed well with simulations based on the orthodox theory. The junction resistance showed thermally activated conduction behaviour at higher temperature. The overall voltage and temperature dependent results show that the transport behaviour of the large arrays of single particle devices obtained by a facile optical lithography and chemical etching process corresponds with the behaviour of single particle devices fabricated by other techniques like e-beam lithography and mechanical breaking methods.     

Creep behavior of INCOLOY alloy 617

The microstructural features of INCOLOY alloy 617 in the solution annealed condition and after long-term creep tests at 700 and 800 °C were characterized and correlated with hardness and creep strength. Major precipitates included (Cr,Mo,Fe)₂₃C₆ carbides and the δ-Ni₃Mo phase. M₆C and MC carbides were also detected within the austenitic grains. However, minor precipitates particularly γ′-Ni₃(Al,Ti) was found to play an important role. At different exposure temperatures, the microstructural features of the Ni–22Cr–12Co–9Mo alloy changed compared with the as-received condition. The presence of discontinuously precipitated (Cr,Mo,Fe)₂₃C₆ carbides and their coarsening until the formation of an intergranular film morphology could be responsible both for a reduction in rupture strength and for enhanced intergranular embrittlement. The fraction and morphology of the γ′-phase, precipitated during exposure to high temperature, also changed after 700 or 800 °C exposure. At the latter test temperature, a lower volume fraction of coarsened and more cubic γ′ precipitates were observed. These microstructural modifications, together with the presence of the δ-phase, detected only in specimens exposed to 700 °C, were clearly responsible for the substantially good creep response observed at 700 °C, compared with that found at 800 °C.     

Gas sensing properties of columnar CeO2 nanostructures prepared by chemical vapor deposition

Columnar CeO2 nanostructures are grown on alumina substrates by a template- and catalyst-free Chemical Vapor Deposition (CVD) approach and subsequently tested as resistive gas sensors of CH3COCH3, H2, NO2. The sensor response is stable and reproducible throughout the whole working temperature range (200-500 degrees C) and directly dependent on the analyte gas and the adopted operating conditions. The higher sensitivity with respect to that displayed by continuous CeO2 thin films demonstrates the potential of fabricating nanostructured sensing devices characterized by improved functional performances.     

New biomimetic constructs for improved in vivo circulation of superparamagnetic nanoparticles

Superparamagnetic iron oxide nanoparticles (SPIOs) have been produced and used as a potent and versatile contrast media for magnetic resonance imaging (MRI). Despite a number of efforts to improve their surface chemistry and biocompatibility, the SPIOs half life in blood circulation is very short and they are rapidly taken up by the reticuloendothelial system (RES). In this paper we describe a new method that permits to avoid the rapid clearance of SPIOs. Nanoparticles are made biocompatible by encapsulation into autologous red blood cells. These biomimetic constructs preserve the main properties of the cells that escape RES clearance as well as the properties of the nanoparticles that perform even better than in blood suspension with reduced T2*. These SPIO-loaded RBCs are promising intravascular imaging contrast agents and could also be addressed to selected body compartments by an external magnetic field.     

Modification of buffered peptone water for improved recovery of heat-injured Salmonella Typhimurium

Abstract: Rapid detection of Salmonella in foods is often limited by the high demand for the sensitivity of detection, poor physiological conditions of the target cells, and high concentration of background flora. In this study, the conditions of nonselective enrichment cultivation were modified in order to improve the quantitative detection of heat‐injured Salmonella in minced meat. The effect of the modifications on the recovery was observed by means of RNA‐based sandwich hybridization, which was adjusted for the quantification of Salmonella enterica 23S rRNA in crude cell extracts. The supplementation of buffered peptone water with the enzyme‐controlled substrate delivery system EnBase‐Flo^® and ferrioxamine E was shown to improve the recovery of cells in both single strain cultures and in the presence of minced meat. The presented results can be used for the development of more efficient enrichment cultivation media for faster detection of food borne Salmonella.     

Chemical composition, in vitro anti-yeast activity and fruit juice preservation potential of lemon grass oil

The anti-yeast activity of lemon grass oil was evaluated against several food spoiling yeasts (Saccharomyces cerevisiae, Zygosaccharomyces bailii, Aureobasidium pullulans, Candida diversa, Pichia fermentans, Pichia kluyveri, Pichia anomala and Hansenula polymorpha) through disc diffusion and microbroth dilution method. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) varied from 0.28 to 1.3 mg/ml and 0.56 to 4.5 mg/ml, respectively, where highest MIC (2.25 mg/ml) was shown by A. pullulans and lowest MIC (0.28 mg/ml) was shown by C. diversa and P. anomala. Kill time assay was conducted for selected three yeast strains where S. cerevisiae showed highest reduction (3 log cfu) in viability within 24 h exposure of lemon grass oil at MFC level. Further, the anti-yeast efficacy of lemon grass oil alone and in combination with thermal treatment was evaluated in real food system i.e. mixed fruit juices. Chemical composition analysis of lemon grass oil by gas chromatography–mass spectrometry (GC–MS) revealed that the dominant compounds were geranial (40.5%), neral (30.7%), geranyl acetate (5.1%), caryophyllene (2.5%). Present results established the superior performance of integrated (thermal–lemon grass oil) treatment over the individual exposure (lemon grass oil or thermal treatment alone) for fruit juice preservation.     

Import-export balance of nitrogen and phosphorus in food, fodder and fertilizers in the Baltic Sea drainage area

Nitrogen (N) and phosphorus (P) are essential elements for life, but in excess they contribute to aquatic eutrophication. The Baltic Sea is a brackish semi-enclosed sea that is heavily influenced by anthropogenic loading of nutrients, resulting in a major environmental problem, eutrophication. In this study, the nutrient balance of the food production and consumption system in seven countries in the Baltic Sea drainage area was quantified for the period 2002-2005. The food production and consumption system accumulates nutrients in the Baltic Sea drainage area, due to extensive imports to the system. The average annual net surplus of nutrients was 1 800 000 tons N and 320 000 tons P in 2002-2005, or annually 28 kg N and 5 kg P per capita. The average total annual import was 2 100 000 tons N and 340 000 tons P during 2002-2005. The largest imports to the system were fertilizers, totaling 1 700 000 tons N and 290 000 tons P. Traded nutrients in food and fodder amounted to a net annual surplus of 180 000 tons N and 25 000 tons P. The nutrient load to the Baltic Sea due to the food consumption and production system was 21% N and 6% P of the respective annual net inputs to the region. This study shows that large amounts of nutrients to Baltic Sea drainage area are inputs from outside the region, eventually contributing to eutrophication. To reduce the nutrient imports, fertilizers should be used more efficiently, nutrients should be recycled more efficiently inside the region, and food system should be guided toward low-nutrient intensive diets.     

Reduction of the olfactory cognitive ability in horses during preslaughter: stress-related hormones evaluation

As horses may perceive several odour signals of danger at slaughter, application of mentholated ointment to their nostrils may limit their perception of danger. To assess the effect of the application of a mentholated ointment to horse nostrils on the stress response during pre-slaughter handling, plasma levels were evaluated for cortisol, beta-endorphin, epinephrine and norepinephrine prior to and after stunning. Twenty draught-type horses were divided into control (n = 10) and treated (n = 10) groups and a mentholated ointment applied to the nostrils of the treated horses following blood sampling in lairage 45 min prior to slaughter. Treatment did not affect plasma concentrations of beta-endorphin or cortisol but significantly reduced the concentrations of epinephrine and norepinephrine observed in post-stun plasma. These results indicated that mentholated ointment applied to the nostrils of horses pre-slaughter reduced their adrenergic response to the slaughter environment, implying that the stress response may be reduced with this technology.