Spectroscopic properties of the photoproducts of pyridoxal-5′-P irradiation: Catalytic site recognition of ribonuclease A

Photoproducts of pyridoxal-5-P, i.e., 4-pyridoxic-5-P and bis-pyridoxal-5-P, have been studied by spectroscopic methods. The spectroscopic properties of bis-pyridoxal-5-P (bis-PLP) resemble those of pyridoxal-5-P (PLP) under similar experimental conditions. The coupling of methylen hydrogens to the phosphorus atom has been shown by NMR spectroscopy. The singlet in the³¹P-NMR spectra and the triplet in¹H-undecoupled experiments confirm the presence of the phosphate group in the 5 position of the structure of the vitamin. The effect of pH and solvent composition on the relative distribution of species of bis-pyridoxine-5-P (bis-PNP) has been investigated by absorption and fluorescence spectroscopy. The acid-base dissociation of the phosphate group is easily detected by emission spectroscopy. Bis-PNP and bis-PLP bind to the enzyme RNase A and they behave as competitive inhibitors with respect to the substrate cytidine-2-3-cyclic phosphate. The natural forms of vitamin B₆, pyridoxine, and pyridoxine-5-P have no effect on the catalytic activity of the protein. Experimental evidence derived from fluorescence and inhibition experiments is consistent with the hypothesis that bis-PNP recognizes the catalytic site of RNase A.     

Laurdan in Fluid Bilayers: Position and Structural Sensitivity

Laurdan (2-dimethylamino-6-lauroylnaphthalene) is a hydrophobic fluorescent probe widely used in lipid systems. This probe was shown to be highly sensitive to lipid phases, and this sensitivity related to the probe microenvironment polarity and viscosity. In the present study, Laurdan was incorporated in 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DPPG), which has a phase transition around 41°C, and DLPC (1,2-dilauroyl-sn-glycero-3-phosphocholine), which is in the fluid phase at all temperatures studied. The temperature dependence of Laurdan fluorescent emission was analyzed via the decomposition into two gaussian bands, a short- and a long-wavelength band, corresponding to a non-relaxed and a water-relaxed excited state, respectively. As expected, Laurdan fluorescence is highly sensitive to DPPG gel–fluid transition. However, it is shown that Laurdan fluorescence, in DLPC, is also dependent on the temperature, though the bilayer phase does not change. This is in contrast to the rather similar fluorescent emission obtained for the analogous hydrophilic probe, Prodan (2-dimethylamino-6-propionylnaphthalene), when free in aqueous solution, over the same range of temperature. Therefore, Laurdan fluorescence seems to be highly dependent on the lipid bilayer packing, even for fluid membranes. This is supported by Laurdan fluorescence anisotropy and spin labels incorporated at different positions in the fluid lipid bilayer of DLPC. The latter were used both as structural probes for bilayer packing, and as Laurdan fluorescence quenchers. The results confirm the high sensitivity of Laurdan fluorescence emission to membrane packing, and indicate a rather shallow position for Laurdan in the membrane.     

Antioxidant Activities of Some New Chromonyl-2,4-Thiazolidinediones and Chromonyl-2,4-Imidazolidinediones Having Chromone Cores

The antioxidant properties of 11 new synthesized chromonyl-2,4-thiazolidinediones and chromonyl-2,4-imidazolidinediones (CBs) were investigated. The antioxidant activities and mechanisms of the CBs interaction with reactive oxygen species (ROS) were clarified using various in vitro antioxidant assay methods including superoxide anion radical ( $ \mathrm{O}\overline{{}_2^{\bullet }} $ ), hydroxyl radical (HO^?), 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH^?) scavenging activity and the iron (II)-ferrozine complex formation. The potassium superoxide/18-crown-6 ether dissolved in dimethylsulfoxide (DMSO) was applied as a source of superoxide anion radical. Hydroxyl radicals were produced in the Fenton-like reaction Fe(II)+H₂O₂. Chemiluminescence, spectrophotometry, and electron paramagnetic resonance (EPR) spectroscopy using 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) as spin trap were applied as the measurement techniques. The CBs examined that exhibited good free radical scavenging activity also showed strong total antioxidant power capacity. Possible mechanisms of antioxidation are proposed to explain the differences in the experimental results between the chromone derivatives with imidazolidine-2,4-dione ring and those with thiazolidine-2,4-dione ring. In conclusion, some of the new CBs are promising to be applied as inhibitors of free radicals.     

Sound absorption and transmission through flexible micro-perforated panels backed by an air layer and a thin plate

This paper describes theoretical and experimental investigations into the sound absorption and transmission properties of micro-perforated panels (MPP) backed by an air cavity and a thin plate. A fully coupled modal approach is proposed to calculate the absorption coefficient and the transmission loss of finite-sized micro-perforated panels-cavity-panel (MPPCP) partitions with conservative boundary conditions. It is validated against infinite partition models and experimental data. A practical methodology is proposed using collocated pressure-velocity sensors to evaluate in an anechoic environment the transmission and absorption properties of conventional MPPCPs. Results show under which conditions edge scattering effects should be accounted for at low frequencies. Coupled mode analysis is also performed and analytical approximations are derived from the resonance frequencies and mode shapes of a flexible MPPCP. It is found that the Helmholtz-type resonance frequency is deduced from the one associated to the rigidly backed MPPCP absorber shifted up by the mass-air mass resonance of the flexible non-perforated double-panel. Moreover, it is shown analytically and experimentally that the absorption mechanisms at the resonances are governed by a large air-frame relative velocity over the MPP surface, with either in-phase or out-of-phase relationships, depending on the MPPCP parameters.     

An authentication case study: Antonio Palomino versus Vicente Guillo paintings in the vaulted ceiling of the Sant Joan del Mercat church (Valencia,Spain)

In 1695, the Valencian artist Vicente Guillo was engaged in painting the vault of the Sant Joan del Mercat church in Valencia, Spain. After preliminary work was carried out, his contract was cancelled. In 1697, Antonio Palomino, renowned for the publication of his technical treatise entitled El Museo Pictorio y Escala Optica, was finally selected as the painter in charge of decorating the vaulted ceiling of Sant Joan del Mercat. This paper reports an analytical study focused on the characterisation and discrimination of the palette and painting procedures used by Palomino and Guillo in the frescoes of Sant Joan del Mercat. For this purpose, Raman spectroscopy combined with light microscopy, scanning electron microscopy–X‐ray microanalysis, voltammetry of microparticles, X‐ray diffraction, Fourier transform infrared spectroscopy and gas chromatography–mass spectrometry has been employed. The use of gypsum as stucco material for the ground layers contrasts with the recommendations made by Palomino in his treatise about the convenience of using slaked lime‐sand mortars according to traditional fresco recipes. Although lead‐based pigments were not traditionally recommended for frescoes because of their empirically known alterations when subjected to strong alkaline fresco medium, both Guillo and Palomino used them. Palomino, probably supported by his personal experience as a painter, recommended and used Naples yellow, which has been found in a good state of preservation. In contrast, white lead areas found on Guillo's paintings have transformed into lead oxides. Other pigments found in the vaulted ceiling such as smalt, goethite, haematite, azurite and malachite have also undergone substantial changes because of the extreme temperature conditions to which they were subjected in the church. Copyright © 2012 John Wiley & Sons, Ltd.     

A closed model category for ()-connected spaces

For each integer , we give a distinct closed model category structure to the category of pointed spaces such that the corresponding localized category is equivalent to the standard homotopy category of -connected CW-complexes. The structure of closed model category given by Quillen to is based on maps which induce isomorphisms on all homotopy group functors and for any choice of base point. For each , the closed model category structure given here takes as weak equivalences those maps that for the given base point induce isomorphisms on for .

     

Acoustic voice analysis in patients with essential tremor

To quantify several acoustic features of the voice in patients with essentialtremor (ET), 28 patients and 28 age- and sex-matched controls were studied. ET severity was assessed with the rating scale for tremor of Fahn, Tolosa, and Marín. The Computerized Speech Lab 4300 program (Kay Elemetrics) was used. Two-second samples of a sustained /a/ and a sentence were captured with a microphone and laryngograph equipment. Measures included fundamental frequency (F₀), frequency perturbation (fitter, Koike algorithm), intensity perturbation (shimmer, Horii algorithm), and harmonic-to-noise ratio (H/N, Yumoto algorithm) of the vowel /a/, and the frequency and intensity variability of the sentence, phonational range, and dynamic range at the natural frequency, maximum phonational time, and s/z ratio. All subjects underwent indirect laryngoscopy and/or laryngeal fibroscopy. When compared with controls, ET patients showed higher jitter, lower H/N ratio (the last one only with laryngographic signal), of the vowel /a/, lower frequency variability in the microphonc signal, lower intensity variability in the laryngographic signal of the sentence, and significantly lower dynamic range at natural frequency of phonation. ET patients reported higher frequency of the presence of high voice intensity, tremor, and struggle. Several acoustic parameters were influenced by the severity of the disease, including shimmer, jitter, H/N ratio, frequency variability of the sentence, and s/z ratio, although neither of the acoustic analysis values or the phonetometric measurements were affected by the presence of voice tremor or by a successful pharmacological treatment of ET.     

Measuring technique for thermal ionisation mass spectrometry of human tracer kinetic study with stable cerium isotopes

Thermal ionisation mass spectrometry (TIMS) method has been developed for the simultaneous detection of different cerium isotopes in biological samples (i.e., blood and urine) at very low concentrations. The work has been done in the frame of a biokinetic study, where different stable cerium isotopes have been administered orally and intravenously as tracers to the human body. In order to develop an appropriate detection method for the tracers in the biological samples, an optimum sample preparation technique has been set and adapted to the specific requirements of the analysis technique used, i.e., TIMS. For sample evaporation and ionisation, the double tantalum filament technique showed the best results. The ions produced were simultaneously collected on a secondary electron multiplier so that the isotopic ratios of the cerium isotopes in the biological samples could be measured. The technique has been optimised for the determination of cerium down to 1?ng loaded on the evaporation filament corresponding to cerium concentrations of down to 1?ng?ml(-1) in the blood or urine samples. It has been shown that the technique is reliable in application and enables studies on cerium metabolism and biokinetics in humans without employing radioactive tracers.     

Acoustic Analysis of Pathological Voices Compressedwith MPEG System

The MPEG-1 Layer 3 compression schema of audio signal, commonly known as mp3, has caused a great impact in recent years as it has reached high compression rates while conserving a high sound quality. Music and speech samples compressed at high bitrates are perceptually indistinguishable from the original samples, but very little was known about how compression acoustically affects the voice signal. A previous work with normal voices showed a high fidelity at high-bitrate compressions both in voice parameters and the amplitude-frequency spectrum. In the present work, dysphonic voices were tested through two studies. In the first study, spectrograms, long-term average spectra (LTAS), and fast Fourier transform (FFT) spectra of compressed and original samples of running speech were compared. In the second study, intensities, formant frequencies, formant bandwidths, and a multidimensional set of voice parameters were tested in a set of sustained phonations. Results showed that compression at high bitrates (96 and 128 kbps) preserved the relevant acoustic properties of the pathological voices. With compressions at lower bitrates, fidelity decreases, introducing some important alterations. Results from both works, Gonzalez and Cervera and this paper, open up the possibility of using MPEG-compression at high bitrates to store or transmit high-quality speech recordings, without altering their acoustic properties.