A preliminary study exploring the mechanical properties of normal and Mgp-deficient mouse femurs during early growth.

Matrix Gla protein (MGP) is mostly known to be a calcification inhibitor, as its absence leads to ectopic calcification of different tissues such as cartilage or arteries. MGP deficiency also leads to low bone mass and delayed bone growth. In the present contribution, we investigate the effect of MGP deficiency on the structural and material mechanical bone properties by focusing on the elastic response of femurs undergoing three-points bending. To this aim, biomechanical tests are performed on femurs issued from Mgp-deficient mice at 14, 21, 28, and 35 days of postnatal life and compared to healthy control femurs. µCT acquisitions enable to reconstruct bone geometries and are used to construct subject-specific finite element models avoiding some of the reported limitations concerning the use of beam-like assumptions for small bone samples. Our results indicate that MGP deficiency may be associated to differences in both structural and material properties of femurs during early stages of development. MGP deficiency appears to be related to a decrease in bone dimensions, compensated by higher material properties resulting in similar structural bone properties at P35. The search for a unique density-elasticity relationship based on calibrated bone mineral density (BMD) indicates that MGP deficiency may affect bone tissue in several ways, that may not be represented uniquely from the quantification of BMD. Despite of its limitation to elastic response, the present preliminary study reports for the very first time the mechanical skeletal properties of Mgp-deficient mice at early stages of development.

Longitudinal relaxation rate measurements for different signals unveiled by nutation spectroscopy: Application to the characterization of two arrangements experienced by water in a clay network.

Nutation consists in monitoring the motion of nuclear magnetization under the application of a radiofrequency (rf) field. With an appropriate amplitude of the rf field, the nutation frequency depends on the NMR relaxation times. This property offers the possibility of differentiating species having the same Larmor frequency but differing by their relaxation times. This may occur for the composite proton NMR signal of water in complex systems. Separate nutation signals are thus observed with the possibility of measuring their longitudinal relaxation times by simply applying a saturation hard pulse, followed by an evolution interval, prior to the nutation sequence. This novel experiment has been used for studying the two sites existing for water in two kaolinite samples (one hydrated after stabilization of several months and the other nonhydrated). It turns out that water in these two sites differs essentially by its transverse relaxation time. Moreover, recovery is surprisingly biexponential for these two signals. A proper analysis of results obtained by this saturation-recovery nutation experiment provides not only the specific longitudinal relaxation rates of water in these two sites but also information about the averaging which occurs at long evolution times. This is discussed with regard of the structure and organization of the clay network. In particular, from relaxation rates at short evolution times, it is shown that this network is mainly constituted of ordered platelets, with a relatively weak proportion of randomly distributed platelets.

Proton nutation spectroscopy. Application to the quantitation of water in a kaolinite sample.

Nutation consists in monitoring the motion of nuclear magnetization under the application of a radio-frequency field. Depending on the amplitude of the rf field, the nutation frequency may be sensitive to the two longitudinal and transverse relaxation rates R1 and R2, hence the possibility of differentiating species having the same resonance frequency in the laboratory frame (the Larmor frequency) but differing by their relaxation rates, as it may occur for the composite proton NMR signal of water in complex systems. Thus, Fourier transform of the nutation curve should provide separate peaks associated with the different species involved in a composite classical NMR signal. As nutation peaks may be close to zero frequency (or even at zero frequency), their full observation requires a complex Fourier transform. This implies a second nutation curve, de-phased by 90° with respect to the first one, achieved here by a second nutation experiment preceded by a 90° hard pulse. Eventually, more accurate parameters are obtained by a non-linear least-squares analysis of the simple nutation experiment. This methodology is applied to water in a natural clay (kaolinite) and reveals the unexpected presence of two peaks which can be characterized by the relaxation rates derived from the line-widths of the nutation signals.

Measurement of short transverse relaxation times by pseudo-echo nutation experiments.

Very short NMR transverse relaxation times may be difficult to measure by conventional methods. Nutation experiments constitute an alternative approach. Nutation is, in the rotating frame, the equivalent of precession in the laboratory frame. It consists in monitoring the rotation of magnetization around the radio-frequency (rf) field when on-resonance conditions are fulfilled. Depending on the amplitude of the rf field, nutation may be sensitive to the two relaxation rates R1 and R2. A full theoretical development has been worked out for demonstrating how these two relaxation rates could be deduced from a simple nutation experiment, noticing however that inhomogeneity of the rf field may lead to erroneous results. This has led us to devise new experiments which are the equivalent of echo techniques in the rotating frame (pseudo spin-echo nutation experiment and pseudo gradient-echo experiment). Full equations of motion have been derived. Although complicated, they indicate that the sum of the two relaxation rates can be obtained very accurately and not altered by rf field inhomogeneity. This implies however an appropriate data processing accounting for the oscillations which are superposed to the echo decays and, anyway, theoretically predicted. A series of experiments has been carried out for different values of the rf field amplitude on samples of water doped with a paramagnetic compound at different concentrations. Pragmatically, as R1 can be easily measured by conventional methods, its value is entered in the data processing algorithm which then returns exclusively the value of the transverse relaxation time. Very consistent results are obtained that way.

Low temperature oxidation of benzene and toluene in mixture with n-decane.

The oxidation of two blends, benzene/n-decane and toluene/n-decane, was studied in a jet-stirred reactor with gas chromatography analysis (temperatures from 500 to 1100 K, atmospheric pressure, stoichiometric mixtures). The studied hydrocarbon mixtures contained 75% of aromatics in order to highlight the chemistry of the low-temperature oxidation of these two aromatic compounds which have a very low reactivity compared to large alkanes. The difference of behavior between the two aromatic reactants is highly pronounced concerning the formation of derived aromatic products below 800 K. In the case of benzene, only phenol could be quantified. In the case of toluene, significant amounts of benzaldehyde, benzene, and cresols were also formed, as well as several heavy aromatic products such as bibenzyl, phenylbenzylether, methylphenylbenzylether, and ethylphenylphenol. A comparison with results obtained with neat n-decane showed that the reactivity of the alkane is inhibited by the presence of benzene and, to a larger extent, toluene. An improved model for the oxidation of toluene was developed based on recent theoretical studies of the elementary steps involved in the low-temperature chemistry of this molecule. Simulations using this model were successfully compared with the obtained experimental results.

New perspectives in the PAW/GIPAW approach: J(P-O-Si) coupling constants, antisymmetric parts of shift tensors and NQR predictions.

In 2001, Pickard and Mauri implemented the gauge including projected augmented wave (GIPAW) protocol for first-principles calculations of NMR parameters using periodic boundary conditions (chemical shift anisotropy and electric field gradient tensors). In this paper, three potentially interesting perspectives in connection with PAW/GIPAW in solid-state NMR and pure nuclear quadrupole resonance (NQR) are presented: (i) the calculation of J coupling tensors in inorganic solids; (ii) the calculation of the antisymmetric part of chemical shift tensors and (iii) the prediction of (14)N and (35)Cl pure NQR resonances including dynamics. We believe that these topics should open new insights in the combination of GIPAW, NMR/NQR crystallography, temperature effects and dynamics. Points (i), (ii) and (iii) will be illustrated by selected examples: (i) chemical shift tensors and heteronuclear (2)J(P-O-Si) coupling constants in the case of silicophosphates and calcium phosphates [Si(5)O(PO(4))(6), SiP(2)O(7) polymorphs and α-Ca(PO(3))(2)]; (ii) antisymmetric chemical shift tensors in cyclopropene derivatives, C(3)X(4) (X = H, Cl, F) and (iii) (14)N and (35)Cl NQR predictions in the case of RDX (C(3)H(6)N(6)O(6)), ß-HMX (C(4)H(8)N(8)O(8)), α-NTO (C(2)H(2)N(4)O(3)) and AlOPCl(6). RDX, ß-HMX and α-NTO are explosive compounds.

Nitrogen-14 nuclear quadrupole resonance (NQR): dramatic sensitivity enhancement by large and fast temperature lowering.

We have observed that, when going rapidly from ambient temperature down to liquid nitrogen temperature, the nitrogen-14 NQR signal (for transitions involving the m=0 spin state, nitrogen-14 being a quadrupolar nucleus of spin I=1) is increased by a factor of ca. 10(2). While Boltzmann statistics cannot explain this enhancement, the strong temperature dependence of the quadrupolar interaction is very likely to be at the origin of this phenomenon. Indeed, the quadrupolar Hamiltonian becomes time dependent and is prone to induce transitions toward the spin state associated with m=0. Its binding and slow relaxing properties result in a durable increased population and consequently in an increased intensity of NQR lines originating from the state m=0.