In vivo quantitative 1H MRS of cerebellum and evaluation of quantitation reproducibility by simulation of different levels of noise and spectral resolution

A quantitative analysis of cerebellar metabolites in normal subjects has been performed by proton MR spectroscopy (MRS) with relaxation time correction. Quantitation was carried out in seven healthy human subjects with the well-established LCModel program. The prior knowledge utilized for quantitation was obtained from solutions containing the major brain metabolites and MRS investigated under the same experimental conditions. The tissue water signal was used as an internal standard for the in vivo studies. Both in vitro (for the prior knowledge template) and in vivo data were acquired separately at 1.5 T by PRESS sequence (TR, 1500 ms; TE, 30 ms). The absolute concentration of main cerebellar metabolites was corrected for relaxation time effects. Different noise and line broadening conditions were considered and simulated in the spectral processing in order to evaluate the effect of spectral quality on the concentration estimates.     

Mn(II) and Zn(II) interactions with peptide fragments from Parkinson's disease genes

Two peptide sequences from PARK9 Parkinson's disease gene, ProAspGluLysHisGluLeu, (P(1)D(2)E(3)K(4)H(5)E(6)L(7)) (1) and PheCysGlyAspGlyAlaAsnAspCysGly (F(1)C(2)G(3)D(4)G(5)A(6)N(7)D(8)C(9)G(10)) (2) were tested for Mn(II), Zn(II) and Ca(II) binding. The fragments are located from residues 1165 to 1171 and 1184 to 1193 in the PARK9 encoded protein. This protein can protect cells from poisoning of manganese, which is an environmental risk factor for a Parkinson's disease-like syndrome. Mono- and bi-dimensional NMR spectroscopy has been used to understand the details of metal binding sites at different pH values and at different ligand to metal molar ratios. Mn(II) and Zn(II) coordination with peptide (1) involves imidazole N(ε) or N(δ) of His(5) and carboxyl γ-O of Asp(2), Glu(3) and Glu(6) residues. Six donor atoms participate in Mn(II) binding resulting in a distorted octahedral geometry, possibly involving bidentate interaction of carboxyl groups; four donor atoms participate in Zn(II) binding resulting in a tetracoordinate geometry. Mn(II) and Zn(II) coordination involves the two cysteine residues with peptide (2); Mn(II) accepts additional ligand bonds from the carboxyl γ-O of Asp(4) and Asp(8) to complete the coordination sphere; the unoccupied sites may contain solvent molecules. The failure of Ca(II) ions to bind to either peptide (1) or (2) appears to result, under our conditions, from the absence of chelating properties in the chosen fragments.     

Microwave-assisted silica-supported aluminum chloride-catalyzed Friedel-Crafts alkylation

Microwave irradiation is a popular method in organic synthesis to achieve high yields in shorter reaction times. This decreases total ‘man-hours’ in a synthetic setting. Another technique used in organic chemistry to decrease manual manipulations, is solid support reagents. The benefits of this approach is that upon completion of a reaction, a simple filtration can be performed which expedites the work-up and also produces less organic waste. Friedel-Crafts alkylation has been explored using microwave chemistry as well as with solid-supported reagents. In comparison with traditional heating, as well as with AlCl₃, superior yields were observed with silica-gel bound aluminum chloride (Si-AlCl_x) when microwave irradiated for only 5 min.     

The study of the mechanical impedance of foods and biomaterials to characterize their linear viscoelastic behavior at high frequencies

A new instrumental setup, which is based on the oscillatory squeezing flow model, was developed to characterize the viscoelastic properties of foods and biopolymers. Analysis of the data was performed by two different approaches. The first approach employed principles of vibration to determine viscous damping and elasticity of the sample harmonically compressed between two plates. The second approach involved the use of additional calculations based on the squeezing flow model which were linked to concepts of vibration analysis, such as mechanical impedance, to determine fundamental rheological parameters like complex viscosity and related (elastic and viscous) moduli. The experimental setup for the method is simple to use and could be attached to existing commercial instruments such as texture analyzers and universal testing machines. The use of the proposed method with this type of instruments would provide them with the additional capability of performing dynamic rheological testing. The dynamic mass of the instrument was significantly low when compared to that of other instruments that use similar principles. This low dynamic mass enabled the use of relatively higher frequencies for the testing of the samples. Comparison of the viscous and elastic moduli obtained with the proposed method and from conventional rheometers for a variety of foods and biopolymers showed good agreements.     

Polyphenols from Citrus Tacle® Extract Endowed with HMGCR Inhibitory Activity: An Antihypercholesterolemia Natural Remedy

Tacle^® is a citrus fruit obtained from the crossbreeding of Clementine and Tarocco cultivars. This fruit retains a promising nutraceutical potential most likely due to a high content in polyphenols, among which the main constituents are the two glycosides naringin and hesperidin. Herein, we evaluated, through an in vitro assay, the capability of Tacle extracts to inhibit the hydroxymethylglutaryl-CoA reductase enzyme, which plays a key role in cholesterol biosynthesis. The results obtained spurred us to investigate whether the anti-enzymatic activity observed may be due to a direct interaction of aglycones naringenin and hesperetin with the enzyme catalytic site. Molecular docking simulations indicated that these two compounds are able to anchor to the protein with binding modes and affinities similar to those found for statins, which represent mainstream medications against hypercholesterolemia. The overall results showed an interesting nutraceutical potential of Tacle, suggesting that its extract could be used for dietary supplementation in the treatment of moderate hypercholesterolemia.     

Advanced diagnostic plan applied to the Royal Villa in Naples

Abstract

To achieve adequate restoration project, the moment of choice of the type of intervention is a very delicate phase that necessarily involves a thorough understanding of the asset in all its aspects, as the location, the history, the architecture, the state of preservation, the pathologies causing the degradation, the degradation in itself, the analysis of constituent materials. The set of all that can be investigated on a product (cultural), to obtaining important information for subsequent maintenance, conservation and/or restoration actions, is what is called a diagnostic plan. In this case it was applied directly on a monument of national importance: the Royal Villa in Naples (today the Municipal Villa). The analysis were performed in situ and in the laboratory (SEM-EDS) on samples taken directly at the Villa, which allowed the creation of a series of analytical cards, containing digital data, required for the selection of expense future of intervention.     

Zinc Interactions with a Soluble Mutated Rat Amylin to Mimic Whole Human Amylin: An Experimental and Simulation Approach to Understand Stoichiometry,Speciation and Coordination of the Metal Complexes

Islet amyloid polypeptide (IAPP) is a hormone co-secreted with insulin and zinc from pancreatic β-cells. To overcome the low solubility of human IAPP, we characterized zinc complexes species formed with 1) a mutated form of rat-IAPP(1–37; R18 H) able to mimic the human IAPP, 2) the r-IAPP(1–37) and the IAPP(1–8) fragment. Stoichiometry, speciation and coordination features of zinc(II) complexes were unveiled by ESI-MS, potentiometry and NMR measurements combined with DFT and free-energy simulations. Mononuclear species start to form around pH 6; Zn²⁺ binds both His18 and N-amino terminus in rat-IAPP(1–37; R18 H). The in silico study allows us to assess not only a structured turn compact domain in r-IAPP(1–37) and r-IAPP(1–37; R18 H) featured by a different free energy barrier for the transition from the compact to elongated conformation upon the coordination of Zn²⁺, but also to bring into light a coordination shell further stabilized by noncovalent interactions.     

Partial cerebral ischemia assessed by "in vivo" 31P NMR spectroscopy in rats.

31P NMR spectroscopy was used to assess the cerebral ischemia status in rats by measuring the relative levels of phosphate metabolites. Partial cerebral ischemia was induced in 49 rats by reversible occlusion of the carotid arteries. Rats were intubated and mechanically ventilated on a hypoxic gas mixture. Physiological parameters such as temperature and arterial pressure were strictly controlled during the experiments. 31P spectra were acquired at 7 T during basal observation, for 15-20 min after the induction of ischemia, and for 1 hr after reperfusion. Depletion and increase in PCr and Pi levels, respectively, were already observable in the collected spectra within few minutes after the onset of ischemia. No appreciable changes were found in the ATP levels.     

Phenolics from Castanea sativa leaves and their effects on UVB-induced damage

The phytochemical investigation of the methanol extract of the leaves of Castanea sativa Mill., source of the Italian PGI (Protected Geographical Indication) product ‘Marrone di Roccadaspide’ (Campania region) afforded as main compounds crenatin (1), chestanin (2), gallic acid (3), cretanin (4), 5-O-p-coumaroylquinic acid (5), p-methylgallic acid (6) and quercetin-3-O-glucoside (7). To quantify the isolated compounds a LC-ESI(QqQ)MS method working with a very sensitive and selective mass tandem experiment called Multiple Reaction Monitoring (MRM) has been developed. Moreover the antioxidant capacity by TEAC assay and the ability of compounds 1–7 to protect HaCaT human keratinocytes from UVB-induced damage has been investigated.