Design, Synthesis and Relaxivity Behaviour of mRNA Targeting Contrast Agents

Introduction: Magnetic Resonance Imaging (MRI) is one of the most important diagnostic tools available in medicine. The specificity and sensitivity of MRI can be further enhanced by the introduction of contrast agents (CAs). As many clinically valuable targets reside inside the cell membrane, the development of efficient intracellular targeted MR contrast agent is required. Potential intracellular targets would be DNA, mRNA or protein/enzymes. The prerequisite for the intracellular targeting is not only the efficient delivery of probes inside the cell but also the colocalization with the target. The objective of the present study is to design and synthesize efficient intracellular MR contrast agents [1] which include three functional domains (i) Gd based MR reporter part (ii) antisense PNA to target mRNA (iii) cell penetrating peptide (CPP) or cholesterol as the delivery agent. The antisense PNA can hybridize uniquely to the complementary mRNA and provides cell specific targeting for cells containing the mRNA. Thus, the expression of the corresponding gene can be visualized non invasively by in vivo MR imaging. Methods: The conjugates were synthesized by continuous solid phase synthesis. Cellular uptake was confirmed by fluorescence microscopy, spectroscopy and MRI of labeled cells. Results: CPP conjugated mRNA targeting CAs were taken up efficiently into cells by an exclusively endosomal mechanism. A subtoxic labeling concentration at low micromolar range was sufficient to enhance significant MR imaging contrast. Cell free binding assays proved a specific interaction with a synthetic target. However, because of the vesicular entrapment, it can be expected that there would be a lack of specific interaction between CA and mRNA located in the cytosol. In order to overcome this problem, cholesterol conjugated CAs are designed and synthesized [2]. The idea behind cholesterol coupling was from the already published report [3] where covalent conjugates of cholesterol and siRNAs were facilitating cellular import and were able to silence protein expression effectively. Initial results have shown that these agents were delivered more efficiently than CPP conjugated CAs. Unfortunately, they were also entrapped in vesicles. Both types of CA were able to enhance contrast in labeled target containing as well as non-targeted parent cells. However, there was not efficient colocalization and specific interaction of CA and target mRNA achievable to be exploited for MR imaging purposes. Conclusion: CPP or lipid coupled CAs are internalized efficiently into cells. However, vesicular entrapment prevented sufficient specific interaction between CA and mRNA. Further modifications are required to achieve the release from endosomes or a direct uptake into the cytosol

New results on source and diffusion spectral features of Galactic cosmic rays: I- B/C ratio

In a previous study (Maurin et al., 2001), we explored the set of parameters describing diffusive propagation of cosmic rays (galactic convection, reacceleration, halo thickness, spectral index and normalization of the diffusion coefficient), and we identified those giving a good fit to the measured B/C ratio. This study is now extended to take into account a sixth free parameter, namely the spectral index of sources. We use an updated version of our code where the reacceleration term comes from standard minimal reacceleration models. The goal of this paper is to present a general view of the evolution of the goodness of fit to B/C data with the propagation parameters. In particular, we find that, unlike the well accepted picture, and in accordance with our previous study, a Kolmogorov-like power spectrum for diffusion is strongly disfavored. Rather, the $\chi^2$ analysis points towards $\delta\gtrsim 0.7$ along with source spectra index $\lesssim 2.0$. Two distinct energy dependences are used for the source spectra: the usual power-law in rigidity and a law modified at low energy, the second choice being only slightly preferred. We also show that the results are not much affected by a different choice for the diffusion scheme. Finally, we compare our findings to recent works, using other propagation models. This study will be further refined in a companion paper, focusing on the fluxes of cosmic ray nuclei.Comment: 32 pages, 13 figures, accepted in A&

Source spectral index of heavy cosmic ray nuclei

From the energy spectra of the heavy nuclei observed by the French-Danish experiment on HEAO-3, the source spectra of the mostly primary nuclei (C, O, Ne, Mg, Si, Ca and Fe) in the framework of an energy dependent leaky box model (Engelmann, et al., 1985) were derived. The energy dependence of the escape length was derived from the observed B/C and sub-iron/iron ratios and the presently available cross sections for C and Fe on H nuclei (Koch-Miramond, et al., 1983). A good fit to the source energy spectra of all these nuclei was obtained by a power law in momentum with an exponent gamma = -2.4+0.05 for the energy range 1 to 25GeV/n (Engelmann, et al., 1985). Comparison with data obtained at higher energy suggested a progressive flattening of these spectra. More accurate spectral indices are sought by using better values of the escape length based on the latest cross section measurements (Webber 1984, Soutoul, et al., this conference). The aim is also to extend the analysis to lower energies down to 0.4GeV/n (kinetic energy observed near Earth), using data obtained by other groups. The only nuclei for which a good data base is possessed in a broad range of energies are O and Fe, so the present study is restricted to these two elements

Charge and energy dependence of the residence time of cosmic ray nuclei below 15 GeV/nucleon

The relative abundance of nuclear species measured in cosmic rays at Earth has often been interpreted with the simple leaky box model. For this model to be consistent an essential requirement is that the escape length does not depend on the nuclear species. The discrepancy between escape length values derived from iron secondaries and from the B/C ratio was identified by Garcia-Munoz and his co-workers using a large amount of experimental data. Ormes and Protheroe found a similar trend in the HEAO data although they questioned its significance against uncertainties. They also showed that the change in the B/C ratio values implies a decrease of the residence time of cosmic rays at low energies in conflict with the diffusive convective picture. These conclusions crucially depend on the partial cross section values and their uncertainties. Recently new accurate cross sections of key importance for propagation calculations have been measured. Their statistical uncertainties are often better than 4% and their values significantly different from those previously accepted. Here, these new cross sections are used to compare the observed B/C+O and (Sc to Cr)/Fe ratio to those predicted with the simple leaky box model

Continuous monitoring of the boundary-layer top with lidar

International audienceContinuous lidar observations of the top height of the boundary layer (BL top) have been performed at Leipzig (51.3° N, 12.4° E), Germany, since August 2005. The results of measurements taken with a compact, automated Raman lidar over a one-year period (February 2006 to January 2007) are presented. Four different methods for the determination of the BL top are discussed. The most promising technique, the wavelet covariance algorithm, is improved by implementing some modifications so that an automated, robust retrieval of BL depths from lidar data is possible. Three case studies of simultaneous observations with the Raman lidar, a vertical-wind Doppler lidar, and accompanying radiosonde profiling of temperature and humidity are discussed to demonstrate the potential and the limits of the four lidar techniques at different aerosol and meteorological conditions. The lidar-derived BL top heights are compared with respective values derived from predictions of the regional weather forecast model COSMO of the German Meteorological Service. The comparison shows a general underestimation of the BL top by about 20% by the model. The statistical analysis of the one-year data set reveals that the seasonal mean of the daytime maximum BL top is 1400 m in spring, 1800 m in summer, 1200 m in autumn, and 800 m in winter at the continental, central European site. BL top typically increases by 100?300 m per hour in the morning of convective days

Applying logic to pulmonary artery catheter use

Mansour and colleagues recommend not routinely using the pulmonary artery catheter to guide hemodynamic management in the intensive care unit, because the perceived benefits are largely intangible [1]. Pulmonary artery catheter monitoring of the right ventricular ejection fraction (RVef) and of the right ventricular end-diastolic volume (EDV), however, reflects powerful yet underutilized relationships that assess right ventricular performance. Since the cardiac output equals the product of the RVef, the EDV and the heart rate, one can assess the RVef to EDV relations as direct measures of right ventricular performance. A series of RVef, EDV and heart rate combinations can give the same cardiac output (Figure 1); monitoring or targeting cardiac output alone ignores this reality. For example, in hypovolemia the EDV is low and the RVef is increased