Thermal response of contrast agent microbubbles: preliminary results from physico-chemical and US-imaging characterization

Hyperthermia (HT) is a therapeutic strategy based on the selective damaging of tumoral cells when heated at temperatures in the range 41-45 degrees C. We are currently investigating the feasibility of Ultrasound (US) imaging to perform a non-invasive, efficient and cost effective temperature monitoring of heated tissues. Commercial US contrast agents (Sonovue, Bracco), consisting in microbubbles of SF(6) coated with a phospholipidic shell, greatly improve the US echo signal from tissues. Further investigations have been performed, consisting in physico-chemical and US-imaging characterization. In conclusion, we demonstrate that Sonovue microbubbles reach their maximal diameter at 40 degrees C, and then a sharp decrease is observed, possible due to the occurrence of gel-sol transition of the phospholipidic shell. At the same temperature the maximal backscattering intensity is predicted and actually experimentally observed. Sonovue, as well as other contrast agents based only on phospholipids, are, therefore, not suitable for use as non-invasive temperature monitoring medium since it is sensitive to temperatures below the hyperthermic range. Although microbubbles are in principle thermally effective, other coating materials should be investigated in order to increase their operative thermal range.     

Commissioning of a multi‐beamline femtoslicing facility at SOLEIL

The investigation of ultrafast dynamics, taking place on the few to sub‐picosecond time scale, is today a very active research area pursued in a variety of scientific domains. With the recent advent of X‐ray free‐electron lasers (XFELs), providing very intense X‐ray pulses of duration as short as a few femtoseconds, this research field has gained further momentum. As a consequence, the demand for access strongly exceeds the capacity of the very few XFEL facilities existing worldwide. This situation motivates the development of alternative sub‐picosecond pulsed X‐ray sources among which femtoslicing facilities at synchrotron radiation storage rings are standing out due to their tunability over an extended photon energy range and their high stability. Following the success of the femtoslicing installations at ALS, BESSY‐II, SLS and UVSOR, SOLEIL decided to implement a femtoslicing facility. Several challenges were faced, including operation at the highest electron beam energy ever, and achievement of slice separation exclusively with the natural dispersion function of the storage ring. SOLEIL's setup also enables, for the first time, delivering sub‐picosecond pulses simultaneously to several beamlines. This last feature enlarges the experimental capabilities of the facility, which covers the soft and hard X‐ray photon energy range. In this paper, the commissioning of this original femtoslicing facility is reported. Furthermore, it is shown that the slicing‐induced THz signal can be used to derive a quantitative estimate for the degree of energy exchange between the femtosecond infrared laser pulse and the circulating electron bunch.     

Low temperature Fourier transform infrared spectra and hydrogen bonding in polycrystalline uracil and thymine

The FTIR spectra of pure NH and isotopically diluted (NH/ND and ND/NH) polycrystalline uracil and thymine were measured in the range 4000-400 cm(-1) at temperatures from 300 to 10K. For the first time, the essentially narrow bands corresponding to the uncoupled stretching (nu(1)) and out of plane bending (nu(4)) NH proton modes of uracil and thymine were observed in the solid phase. It was found that in the nu(4) region the spectra reveal more details on the H-bond interactions present in both solids than in the nu(1) range. The frequencies of the various bands observed in both spectral regions were used for estimation of the H-bond energy, using empirical correlations between this property and both the red shift of nu(1) and the blue shift of nu(4) that occur upon crystallization due to the establishment of the H-bonds. The results are compared with known thermodynamic, structural and theoretical data. The IR data also suggest that the H-bond networks of both crystals contain, besides the two NH...O=C bonds revealed by X-ray experiments, additional types of H-bonds, which do not show long range periodicity and, thus, cannot be detected by the conventional structural methods. The assignment of some other bands in the spectra of both substances was also reviewed.     

DFT/TDDFT investigation of the stepwise deprotonation in tetracycline: pK<Subscript>a</Subscript> assignment and UV–vis spectroscopy

Tetracyclines are a class of derivatives of polycyclic naphthacene carboxamide, which have attracted wide interest in the pharmaceutical field for their use as antibiotics. These molecules are characterized by a substantial conformational flexibility and by the presence of different binding sites which endow tetracycline with a noticeable capability in binding biological targets. A salient property of tetracyclines is the presence of multiple acidic groups: four equilibrium constants have been measured for the fully protonated tetracycline (TCH₃ ⁺) but so far no clear information concerning the pK_as of the various sites has been reported. We present here a computational investigation on the correlation between the acid–base and the spectroscopic properties of this important class of compounds. Starting from the TCH₃ ⁺ species, the pK_a of all the possible deprotonation sites has been computed by DFT calculations. The computed pK_as nicely compare with the experimental data, within 1 pK_a unit, allowing us to individuate the products of the first deprotonation. This procedure has been iteratively repeated using as starting species the products singled out from the previous deprotonation, thus individuating the stepwise products of each deprotonation step. Then, the optical absorption spectra have been computed for all the species involved in the protonation/deprotonation equilibria, comparing the results with the experimental data. The good agreement between theory and experiment has allowed us to rationalize the correlation between the solution pH and the absorption spectra.     

Complexes of tetracyclines with divalent metal cations investigated by stationary and femtosecond-pulsed techniques

Spectroscopic techniques both in steady-state (in absorption and emission) and pulsed (absorption of excited states with femtosecond resolution) conditions were used to study the complexation process between six molecules belonging to the tetracycline family and Mg(2+); in the case of TC the study was extended to the metal ions Ca(2+) and Cu(2+). The study was carried out in aqueous solution at various pH values, where one acid-base form of the substrate prevails over the others. The processing of experimental results, performed by means of Singular Value Decomposition and Global Analysis methods, allowed us to evaluate the extent of interaction through the association constants, to identify the number of equilibria present in solution and the stoichiometry (1:1 or 1:2) of the tetracycline:metal ion complex, and to define the spectral and photophysical properties of the latter (in terms of fluorescence quantum yields, lifetimes and rate constants). In fact, the (allowed) radiative decay process is a minor root for the lowest excited state of the complexes which mainly decay to the ground state by internal conversion. Details of the complexation sites are proposed for the various protonated forms of tetracyclines, and for the various cations in the case of TC. In particular, the molecular structure seems to affect significantly the dynamics of interaction when the upper peripheral region of tetracycline is rich in additional hydroxyl groups. Moreover, the state of protonation of the substrate produces changes in the order of the complexation sites, whose affinity for the cation increases significantly when they are negatively charged owing to the loss of protons. Magnesium and calcium (hard cations) give similar interactions, at least in acid solution, while copper(ii) (borderline cation) binds more efficiently on different sites, thus forming complexes with different properties.     

AFM-based force-clamp monitors lipid bilayer failure kinetics

The lipid bilayer rupture phenomenon is here explored by means of atomic force microscopy (AFM)-based force clamp, for the first time to our knowledge, to evaluate how lipid membranes respond when compressed under an external constant force, in the range of nanonewtons. Using this method, we were able to directly quantify the kinetics of the membrane rupture event and the associated energy barriers, for both single supported bilayers and multibilayers, in contradistinction to the classic studies performed at constant velocity. Moreover, the affected area of the membrane during the rupture process was calculated using an elastic deformation model. The elucidated information not only contributes to a better understanding of such relevant process, but also proves the suitability of AFM-based force clamp to study model structures as lipid bilayers. These findings on the kinetics of lipid bilayers rupture could be extended and applied to the study of other molecular thin films. Furthermore, systems of higher complexity such as models mimicking cell membranes could be studied by means of AFM-based force-clamp technique.     

Efficient γ-amino-proline-derived cell penetrating peptide-superparamagnetic iron oxide nanoparticle conjugates via aniline-catalyzed oxime chemistry as bimodal imaging nanoagents

Aniline-catalyzed oxime chemistry was employed to conjugate a γ-amino-proline-derived cell penetrating peptide to superparamagnetic iron oxide nanoparticles (SPIONs). Internalization of the novel nanoconjugate into HeLa cells was found to be remarkably higher compared to the analogous TAT-SPION conjugate.     

Photoinduced electron transfer in Os(terpyridine)-biphenylene-(bi)pyridinium assemblies

A series of linearly arranged donor-spacer-acceptor (D-S-A) systems 1-3, has been prepared and characterized. These dyads combine an Os(II)bis(terpyridine) unit as the photoactivable electron donor (D), a biphenylene (2) or phenylene-xylylene (3) fragment as the spacer (S), and a N-aryl-2,6-diphenylpyridinium electrophore (with aryl = 4-pyridyl or 4-pyridylium in 1 or 2/3, respectively) as the acceptor (A). Their absorption spectra, redox behavior, and luminescence properties (both at 77 K in rigid matrix and at 298 K in fluid solution) have been studied. The electronic structure and spectroscopic properties of a representative compound of the series (i.e., 2) have also been investigated at the theoretical level, performing Density Functional Theory (DFT)-based calculations. Time-dependent transient absorption spectra of 1-3 have also been recorded at room temperature. The results indicate that efficient photoinduced oxidative electron transfer takes place in the D-S-A systems at room temperature in fluid solution, for which rate constants (in the range 4 × 10(8)-2 × 10(10) s(-1)) depend on the driving force of the process and the spacer nature. In all the D-S-A systems, charge recombination is faster than photoinduced charge separation, in spite of the relatively large energy of the D(+)-S-A(-) charge-separated states (between 1.47 and 1.78 eV for the various species), which would suggest that the charge recombination occurs in the Marcus inverted region. Considerations based on superexchange mechanism suggest that the reason for the fast charge recombination is the presence of a virtual D-S(+)-A(-) state at low energy--because of the involvement of the easily oxidizable biphenylene spacer--which is beneficial for charge recombination via superexchange but unsuitable for photoinduced charge separation. To further support the above statement, we prepared a fourth D-S-A species, 4, analogous to 2 but with a (hardly oxidizable) single phenylene fragment serving as the spacer. For such a species, charge recombination (about 3 × 10(10) s(-1)) is slower than photoinduced charge separation (about 1 × 10(11) s(-1)), thereby confirming our suggestions.     

Macrophage Migration Inhibitory Factor (MIF) and Its Homologue D-Dopachrome Tautomerase (DDT) Inversely Correlate with Inflammation in Discoid Lupus Erythematosus

Discoid Lupus Erythematosus (DLE) is a chronic cutaneous disease of unknown etiology and of immunoinflammatory origin that is characterized by inflammatory plaques and may lead to disfiguring scarring and skin atrophy. Current treatments are limited, with a large proportion of patients either poorly or not responsive, which makes DLE an unmet medical need. Macrophage migration inhibitory factor (MIF) is the prototype of a pleiotropic family of cytokine that also includes the recently discovered homologue D-dopachrome tautomerase (DDT) or MIF2. MIF and DDT/MIF-2 exert several biological properties, primarily, but not exclusively of a proinflammatory nature. MIF and DDT have been suggested to play a key role in the pathogenesis of several autoimmune diseases, such as multiple sclerosis and type 1 diabetes, as well as in the development and progression of certain forms of cancers. In the present study, we have performed an immunohistochemistry analysis for the evaluation of MIF in DLE lesions and normal skin. We found high levels of MIF in the basal layer of the epidermis as well as in the cutaneous appendage (eccrine glands and sebocytes) of normal skin. In DLE lesions, we observed a significant negative correlation between the expression of MIF and the severity of inflammation. In addition, we performed an analysis of MIF and DDT expression levels in the skin of DLE patients in a publicly available microarray dataset. Interestingly, while these in silico data only evidenced a trend toward reduced levels of MIF, they demonstrated a significant pattern of expression and correlation of DDT with inflammatory infiltrates in DLE skins. Overall, our data support a protective role for endogenous MIF and possibly DDT in the regulation of homeostasis and inflammation in the skin and open up novel avenues for the treatment of DLE.     

Posture as a chaotic system and an application to the Parkinson’s disease

In this work we investigate and compare a number of time series of stabilograms of healthy subjects and Parkinsonians. This is carried out by means of the chaos paradigm through the preliminary computation of the first minimum of the mutual information function and the embedding dimension (using false nearest neighbours) in order to obtain the correlation dimension as well as the largest Lyapunov exponent. We show that the postural act is indeed chaotic and especially that the latter two parameters do not allow to discriminate healthy subjects from parkinsonians. Moreover we report a discrepancy of our values with those found in previous works.