Diphenylene Iodonium Is a Noncovalent MAO Inhibitor: A Biochemical and Structural Analysis

Diphenylene iodonium (DPI) is known for its inhibitory activities against many flavin- and heme-dependent enzymes, and is often used as an NADPH oxidase inhibitor. We probed the efficacy of DPI on two well-known drug targets, the human monoamine oxidases MAO A and B. UV-visible spectrophotometry and steady-state kinetics experiments demonstrate that DPI acts as a competitive and reversible MAO inhibitor with K_i values of 1.7 and 0.3 μM for MAO A and MAO B, respectively. Elucidation of the crystal structure of human MAO B bound to the inhibitor revealed that DPI binds deeply in the active-site cavity to establish multiple hydrophobic interactions with the surrounding side chains and the flavin. These data prove that DPI is a genuine MAO inhibitor and that the inhibition mechanism does not involve a reaction with the reduced flavin. This binding and inhibitory activity against the MAOs, two major reactive oxygen species (ROS)-producing enzymes, will have to be carefully considered when interpreting experiments that rely on DPI for target validation and chemical biology studies on ROS functions.     

Flow cytometric assessment of the antimicrobial activity of essential oils against Listeria monocytogenes

Flow cytometry was applied to assess the antimicrobial activity of oregano, thyme and cinnamon essential oils against Listeria monocytogenes ATCC19114, using combined staining with propidium iodide (PI) for membrane damage evaluation and carboxyfluorescein diacetate (cFDA) for esterase activity detection. The antimicrobial activity of the essential oils was also tested at different NaCl concentrations.Significant differences were observed between plate count results and flow cytometric data, which suggested the presence of a sublethally stressed subpopulation, not able to form colonies on agar plates.Following treatments, flow cytometric assessment clearly discriminated three different subpopulations: viable, dead and injured cells. Cinnamon essential oil exerted a different impact on the cellular subpopulations, with a lower overall activity and a large percentage of cells having minimally damaged membranes. On the contrary, membrane disintegration seemed to be the primary inactivation mechanism of oregano and thyme essential oils.The antimicrobial activity of the essential oils increased with NaCl concentration increase, but higher NaCl concentrations were necessary following treatments with cinnamon essential oil.Our findings suggest differences in the mode of action of cinnamon essential oil against L. monocytogenes, in comparison with thyme and oregano essential oils.     

Effect of tomato by-products in the diet of Comisana sheep on composition and conjugated linoleic acid content of milk fat

To evaluate the effect of supplementing the diet of Comisana sheep with by-products from industrial tomato manufacture on the fat composition and conjugated linoleic acid (CLA) content of milk fat, two groups of 50 ewes each were fed either total mixed ration standard (TMRS) or total mixed ration with added tomato by-products (TMRA). Milk fat composition was determined by high-resolution gas chromatography (HRGC). The milk fat content for the animals fed the TMRA diet increased by 6.41% (P < 0.01) after six weeks, compared with the animals fed the TMRS diet. The CLA content in the milk fat for the group of animals fed the TMRA diet was 19.8% (P < 0.05) higher than for those fed the TMRS diet, and reached 1.33 g 100 g^?1 fat, while the polyunsaturated fatty acid (PUFA) content increased by a 6.43% (P < 0.05) and reached 7.12 g 100 g^?1 fat. The fatty acid composition showed an increase in the amount of polyunsaturated fatty acids. The n ? 3:n ? 6 ratio increased by 13% in the milk from sheep fed with the TMRA diet. These observations were confirmed by triglyceride analysis, which showed a decrease in the amount of short-chain (C₂₈–C₃₂) and medium-chain (C₃₄–C₄₂) triglycerides after six weeks, while the opposite was observed for the long-chain triglycerides (C₄₄–C₅₄).     

Second Harmonic Generation as a Tool for Measuring Mode Coupling,Pulse Length and Mode Size in Short Laser Pulses

Abstract

We report on a detailed analysis of second harmonic generation with a multi-modal beam for a non-collinear geometry. As a result some improvements for a picosecond pulse width analyser are suggested. Experimental results relative to both a free electron laser and to a mode locked Nd: YAG laser are discussed. This modified version of the pulse analyser is able to monitor, in real time, the relative pulse-to-pulse change of mode size, pulse length and mode mixing effect independently of the pulse length from femtoseconds to milliseconds.     

Indigenous Saccharomyces cerevisiae strains and their influence on the quality of Cataratto, Inzolia and Grillo white wines

The present paper deals with three new strains of Saccharomyces cerevisiae, isolated in old wineries of Sicily, which were microbiologically and molecularly characterized and tested for their ability to produce white wines. Examined in terms of their growth pattern, fermentation vigour, sulphite tolerance, fermentative power, spore formation, and production of acetic acid, hydrogen sulphide and phenolic off-flavours, the strains were utilized as starters in experimental fermentations of musts obtained from the cultivars Inzolia, Grillo and Catarratto. Further, the three musts were also fermented using two commercial S. cerevisiae strains. The quality of the wines produced was confirmed by their principal oenochemical parameters, by sensory analysis and qualitative and quantitative determination of the volatile aroma constituents. All the data were statistically elaborated. Interestingly, the new selected yeasts were able to increase the pear notes (Z)-ethyl-4-decenoate, (E)-ethyl-3-decenoate, and (Z)-ethyl-3-decenoate which are fundamental for the aroma of these Sicilian wines. From our results, the new yeast strains were found to produce white wines of a quality which was not inferior to those obtainable with the best commercial strains selected in other geographical areas, but also with a distinctive aromatic profile.     

Polarized wave coupling and selection rules violation inalkali-halide crystals with FA centers

Self-diffraction of light has been investigated in KCl with F_A centers in order to elucidate the role of F_A-center kinetics in holographic recording under resonant optical excitation. It is shown that diffraction efficiency for holographic grating recorded in the spectral range of the F_A1 transition can be substantially increased by populating the orientations which immediately interact with the polarized light by means of lateral illumination of the crystal with an auxiliary beam. The optimum range of auxiliary beam intensities leading to improvement of grating quality is found. Such a range is directly defined by the parameter χ, characterizing the selection rules violation for F_A center in KCl. It has been found that the experimentally determined value χ=0.04 is consistent with the self-diffraction data obtained in this paper     

Characterization and fractionation of phenolic compounds extracted from olive oil mill wastewaters

Liquid–liquid extraction was used in order to recover phenolic compounds from olive mill wastewater (OMWW), a polluting by-product of olive oil production process, and the extraction parameters have been optimized. HPLC analysis showed that hydroxytyrosol is the most abundant biophenol in ethyl acetate extracts from an acidified OMWW. Many other biophenols (tyrosol, caffeic acid, vanillic acid, verbascoside, luteolin-7-glucoside, dialdehydic form of decarboxymethyl oleuropein aglycon, ligstroside, luteolin) were identified.     

Nidesy,a laboratory prototype System for determining nitrogen concentration in metal bath continuously

A laboratory prototype of nitrogen determination system (Nidesy) for measurements in molten iron is developed. The method is based on the exchange of nitrogen from the molten iron to a gas phase, the transformation of this gaseous nitrogen in nitrogen oxides and the measurement of produced NO₂ by a suitable sensor. The measurement apparatus consists of three mains parts: first, an apparatus that puts the liquid and gaseous phase in contact and conveys the gas bubbled from the bath to a reactor: blower probe; second, a reactor that transforms N₂ of the blown gas into NO₂: reactor; third, an electrochemical sensor for NO₂ determination: NO₂ sensor. The laboratory experimental activity consists of a step of optimisation of the engineering and operative aspects of whole apparatus; as a result, an instrumental configuration with high sensitivity allowing high accuracy and precision has been obtained, as well as creation and use of the calibration curve (ΔEMF vs dissolved N) for measuring in liquid iron at 1650°C. On the basis of the obtained results a Nidesy for both laboratory and industrial use can be envisaged, for example in a kinetic study of nitrogen absorption/desorption, or in a steel plant, such as a tundish or a continuous casting mould     

Optimization of Picrosirius red staining protocol to determine collagen fiber orientations in vaginal and uterine cervical tissues by Mueller polarized microscopy

Polarized microscopy provides unique information on anisotropic samples. In its most complete implementation, namely Mueller microscopy, this technique is well suited for the visualization of fibrillar proteins orientations, with collagen in the first place. However, the intrinsic optical anisotropy of unstained tissues has to be enhanced by Picrosirius Red (PR) staining to enable Mueller measurements. In this work, we compared the orientation mapping provided by Mueller and second harmonic generation (SHG) microscopies on PR stained samples of vaginal and uterine cervix tissues. SHG is a multiphoton technique that is highly specific to fibrillar collagen, and was taken as the “gold standard” for its visualization. We showed that Mueller microscopy can be safely used to determine collagen orientation in PR stained cervical tissue. In contrast, in vaginal samples, Mueller microscopy revealed orientations not only of collagen but also of other anisotropic structures. Thus PR is not fully specific to collagen, which necessitates comparison to SHG microscopy in every type of tissue. In addition to this study of PR specificity, we determined the optimal values of the staining parameters. We found that staining times of 5 min, and sample thicknesses of 5 µm were sufficient in cervical and vaginal tissues. Microsc. Res. Tech. 78:723–730, 2015. © 2015 Wiley Periodicals, Inc.