81 research outputs found

    Investigation of genomic DNA methylation by ultraviolet resonant Raman spectroscopy

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    Cytosine plays a preeminent role in DNA methylation, an epigenetic mechanism that regulates gene expression, the misregulation of which can lead to severe diseases. Several methods are nowadays employed for assessing the global DNA methylation levels, but none of them combines simplicity, high sensitivity, and low operating costs to be translated into clinical applications. Ultraviolet (UV) resonant Raman measurements at excitation wavelengths of 272 nm, 260 nm, 250 nm, and 228 nm have been carried out on isolated deoxynucleoside triphosphates (dNTPs), on a dNTP mixture as well as on genomic DNA (gDNA) samples, commercial from salmon sperm and non-commercial from B16 murine melanoma cell line. The 228 nm excitation wavelength was identified as the most suitable energy for enhancing cytosine signals over the other DNA bases. The UV Raman measurements performed at this excitation wavelength on hyper-methylated and hypo-methylated DNA from Jurkat leukemic T-cell line have revealed significant spectral differences with respect to gDNA isolated from salmon sperm and mouse melanoma B16 cells. This demonstrates how the proper choice of the excitation wavelength, combined with optimized extraction protocols, makes UV Raman spectroscopy a suitable technique for highlighting the chemical modifications undergone by cytosine nucleotides in gDNA upon hyper- and hypo-methylation events

    Short-wavelength four wave mixing experiments using single and two-color schemes at FERMI

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    The development of ultra-bright extreme ultraviolet (EUV) and X-ray free electron laser (FEL) sources has enabled the extension of wave-mixing approaches into the short wavelength regime. Such a class of experiments relies upon nonlinear interactions among multiple light pulses offering a unique tool for exploring the dynamics of ultrafast processes and correlations between selected excitations at relevant length and time scales adding elemental and site selectivity as well. Besides the availability of a suitable photon source, the implementation of wave mixing methodology requires efforts in developing the instrumental set-up. We have realized at the FERMI FEL two dedicated set-ups to handle multiple FEL beams with preselected parameters in a non-collinear fashion and control their interaction sequence at the target. These unique apparatuses, combined with the exceptional characteristics of the seeded FERMI FEL, have allowed us to make the first steps into this field and further advances are foreseen in the near future

    Probing the molecular connectivity of water confined in polymer hydrogels

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    The molecular connectivity and the extent of hydrogen-bond patterns of water molecules confined in the polymer hydrogels, namely cyclodextrin nanosponge hydrogels, are here investigated by using vibrational spectroscopy experiments. The proposed spectroscopic method exploits the combined analysis of the vibrational spectra of polymers hydrated with water and deuterated water that allows us to separate and selectively investigate the temperature-evolution of the HOH bending mode of engaged water molecules and of the vibrational modes assigned to specific chemical groups of the polymer matrix involved in the physical interactions with water. As main results, we find a strong experimental evidence of a liquid-like behaviour of water molecules confined in the nano-cavities of hydrogel and we observe a characteristic destructuring effect on the hydrogen-bonds network of confined water induced by thermal motion. More interestingly, the extent of this temperature-disruptive effect is found to be selectively triggered by the cross-linking degree of the hydrogel matrix. These results give a more clear picture of the molecular mechanism of water confinement in the pores of nanosponge hydrogel and open the possibility to exploit the spectroscopic method here proposed as investigating tools for water-retaining soft materials

    Tunability experiments at the FERMI@Elettra free-electron laser

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    FERMI@Elettra is a free electron-laser (FEL)-based user facility that, after two years of commissioning, started preliminary users' dedicated runs in 2011. At variance with other FEL user facilities, FERMI@Elettra has been designed to deliver improved spectral stability and longitudinal coherence. The adopted scheme, which uses an external laser to initiate the FEL process, has been demonstrated to be capable of generating FEL pulses close to the Fourier transform limit. We report on the first instance of FEL wavelength tuning, both in a narrow and in a large spectral range (fine- and coarse-tuning). We also report on two different experiments that have been performed exploiting such FEL tuning. We used fine-tuning to scan across the 1s–4p resonance in He atoms, at ≈23.74 eV (52.2 nm), detecting both UV–visible fluorescence (4p–2s, 400 nm) and EUV fluorescence (4p–1s, 52.2 nm). We used coarse-tuning to scan the M4,5 absorption edge of Ge (∼29.5 eV) in the wavelength region 30–60 nm, measured in transmission geometry with a thermopile positioned on the rear side of a Ge thin foil

    The anterior tarsal syndrome. Report of four cases.

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    Early somatosensory evoked potentials in supratentorial brain tumors.

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    Early somatosensory evoked potentials were recorded in 33 patients with supratentorial brain tumors (9 benign tumors, 17 glioblastomas and 7 metastases). All the cases were studied with CT scan and all but 2 were surgically explored and histologically classified. Evoked potentials were statistically analyzed regarding the nature and site of the tumors. The temporal malignant tumors showed the most significant alterations in latencies, particularly for waves P25, N34 and P44

    Hydration properties and water structure in aqueous solutions of native and modified cyclodextrins by UV Raman and Brillouin scattering

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    We have studied aqueous solutions of native and chemically modified cyclodextrins (CDs) by means of UV Raman and Brillouin scattering. Analysis of the spectral profile of the OH\u2010stretching Raman signal, which is sensitive to the intermolecular organization of water, reveals a remarkable reduction of the population of ordered tetrahedral water structures inside the hydration shell of substituted CDs. As a remarkable result, this destructuring effect seems to be mainly related to the number of substituted hydroxyl groups in the CD ring rather than to the chemical nature of the substituent group. UV Brillouin scattering experiments confirm the structural picture emerging from the UV Raman study, also providing an estimate of the activation energy associated to the collective H\u2010bond restructuring mechanism in CD solutions. Overall, the results provide a coherent description of the water\u2013solute interactions in aqueous solutions of CDs
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