Quantitative Proteomic Approach Reveals Altered Metabolic Pathways in Response to the Inhibition of Lysine Deacetylases in A549 Cells under Normoxia and Hypoxia

Growing evidence is showing that acetylation plays an essential role in cancer, but studies on the impact of KDAC inhibition (KDACi) on the metabolic profile are still in their infancy. Here, we analyzed, by using an iTRAQ-based quantitative proteomics approach, the changes in the proteome of KRAS-mutated non-small cell lung cancer (NSCLC) A549 cells in response to trichostatin-A (TSA) and nicotinamide (NAM) under normoxia and hypoxia. Part of this response was further validated by molecular and biochemical analyses and correlated with the proliferation rates, apoptotic cell death, and activation of ROS scavenging mechanisms in opposition to the ROS production. Despite the differences among the KDAC inhibitors, up-regulation of glycolysis, TCA cycle, oxidative phosphorylation and fatty acid synthesis emerged as a common metabolic response underlying KDACi. We also observed that some of the KDACi effects at metabolic levels are enhanced under hypoxia. Furthermore, we used a drug repositioning machine learning approach to list candidate metabolic therapeutic agents for KRAS mutated NSCLC. Together, these results allow us to better understand the metabolic regulations underlying KDACi in NSCLC, taking into account the microenvironment of tumors related to hypoxia, and bring new insights for the future rational design of new therapies.     

Local residence time,residence revolution,and residence volume distributions in twin‐screw extruders

This work was aimed at studying the overall, partial, and local residence time distributions (RTD); overall, partial and local residence revolution distributions (RRD) and overall, partial and local residence volume distributions (RVD) in a co‐rotating twin screw extruder, on the one hand; and establishing the relationships among them, on the other hand. Emphasis was placed on the effects of the type and geometry of mixing elements (a gear block and various types of kneading elements differing in staggering angle) and process parameters on the RTD, RRD and RVD. The overall and partial RTD were directly measured in‐line during the extrusion process and the local ones were calculated by deconvolution based on a statistical theory. The local RTD allowed comparing the mixing performance of mixing elements. Also it was confirmed both experimentally and theoretically that specific throughput, defined as a ratio of throughput (Q) over screw speed (N), controlled all the above three types of residence distributions, be they local, partial or overall. The RRD and RVD do not provide more information on an extrusion process than the corresponding RTD. Rather they are different ways of representing the same phenomena. POLYM. ENG. SCI., 48:19–28, 2008. © 2007 Society of Plastics Engineers     

JaTS: a fully portable seismic tomography software based on Fresnel wavepaths and a probabilistic reconstruction approach

JaTS, a Java 2D seismic tomography software, is presented. It implements original algorithms achieving optimal accuracy with reasonable computing costs. A second-order Fast Marching Method (FMM) is used for solving the eikonal equation, therefore enabling a fast and robust computation of seismic traveltimes between sources and receivers. The wavepaths are materialized by Fresnel volumes rather than by conventional rays. This approach accounts for complex velocity models and has the advantage of considering the effects of the wave frequency in the velocity model resolution. The model is computed by a Simultaneous Iterative Reconstruction Technique (SIRT) which has been reformulated to integrate Fresnel wavepaths by using a probabilistic approach. In addition, various utilities are implemented, such as a tapering filter, used to decrease artifact effects occurring in the vicinity of the sources and receivers. The software also offers the possibility of reconstructing the velocity field on a grid larger than the one used for the wave propagation computation. This contributes to stabilize the estimated values. All of the seismic processing tools have been integrated with a user-friendly graphical interface. JaTS represents a tightly integrated tool suite that supports the entire process of importing the SG2 field records, first-break picking, forward modeling and velocity-field computing across multiple platforms.     

Li-intercalation behaviour of vanadium oxide thin film prepared by thermal oxidation of vanadium metal

In order to produce thin films of crystalline V₂O₅, vanadium metal was thermally oxidised at 500 °C under oxygen pressures between 250 and 1000 mbar for 1-5 min. The oxide films were characterised by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), X-ray diffraction (XRD) and Rutherford backscattering spectrometry (RBS). The lithium intercalation performance of the oxide films was investigated by cyclic voltammetry (CV), chronopotentiometry and electrochemical impedance spectroscopy (EIS). It was shown that the composition, the crystallinity and the related lithium intercalation properties of the thin oxide films were critically dependent on the oxidation conditions. The formation of crystalline V₂O₅ films was stimulated by higher oxygen pressure and longer oxidation time. Exposure for 5 min at 750 mbar O₂ at 500 °C resulted in a surface oxide film composed of V₂O_5, and consisting of crystallites up to 200 nm in lateral size. The thickness of the layer was about 100 nm. This V₂O₅ oxide film was found to have good cycling performance in a potential window between 3.8 and 2.8 V, with a stable capacity of 117 ± 10 mAh/g at an applied current density of 3.4 μA/cm². The diffusion coefficients corresponding to the two plateaus at 3.4 and 3.2 V were determined from the impedance measurements to (5.2 and 3.0) × 10⁻¹³ cm² s⁻¹, respectively. Beneath the V₂O₅ layer, lower oxides (mainly VO₂) were found close to the metal. At lower oxygen pressure and shorter exposure times, the oxide films were less crystalline and the amount of V⁴⁺ increased in the surface oxide film, as revealed by XPS. At intermediate oxygen pressures and exposure times a mixture of crystalline V₂O₅ and V₆O₁₃ was found in the oxide film.     

Embryonal metabolism

It is very difficult to have a clear and homogeneous idea of the embryo metabolism. In fact it may vary from one species to another and also according to the embryonic stage: i.e. before and after genomic activation. Basic compounds such as glucose may be toxic, but obviously, it is more the problem of the quantity introduced in the culture media and an unsuitable balance between the metabolites which may impair the embryonic development. At low concentration glucose is actively metabolised by embryos. High levels of amino acids are deleterious (due to release of ammonia), but they are necessary at low concentrations. Addition of serum or other biological fluids is generally useless. Further knowledge on embryo metabolism is necessary to avoid culture medium related delay or developmental blocks. Sequential media are at least partly the answer.     

Development of Covalent Chitosan-Polyethylenimine Derivatives as Gene Delivery Vehicle: Synthesis,Characterization, and Evaluation

There is an increasing interest in cationic polymers as important constituents of non-viral gene delivery vectors. In the present study, we developed a versatile synthetic route for the production of covalent polymeric conjugates consisting of water-soluble depolymerized chitosan (dCS; M_W 6–9 kDa) and low molecular weight polyethylenimine (PEI; 2.5 kDa linear, 1.8 kDa branched). dCS-PEI derivatives were evaluated based on their physicochemical properties, including purity, covalent bonding, solubility in aqueous media, ability for DNA condensation, and colloidal stability of the resulting polyplexes. They were complexed with non-integrating DNA vectors coding for reporter genes by simple admixing and assessed in vitro using liver-derived HuH-7 cells for their transfection efficiency and cytotoxicity. Using a rational screening cascade, a lead compound was selected (dCS-Suc-LPEI-14) displaying the best balance of biocompatibility, cytotoxicity, and transfection efficiency. Scale-up and in vivo evaluation in wild-type mice allowed for a direct comparison with a commercially available non-viral delivery vector (in vivo-jetPEI). Hepatic expression of the reporter gene luciferase resulted in liver-specific bioluminescence, upon intrabiliary infusion of the chitosan-based polyplexes, which exceeded the signal of the in vivo jetPEI reference formulation by a factor of 10. We conclude that the novel chitosan-derivative dCS-Suc-LPEI-14 shows promise and potential as an efficient polymeric conjugate for non-viral in vivo gene therapy.     

Cardiac Oxidative Signaling and Physiological Hypertrophy in the Na/K-ATPase α1s/sα2s/s Mouse Model of High Affinity for Cardiotonic Steroids

The Na/K-ATPase is the specific receptor for cardiotonic steroids (CTS) such as ouabain and digoxin. At pharmacological concentrations used in the treatment of cardiac conditions, CTS inhibit the ion-pumping function of Na/K-ATPase. At much lower concentrations, in the range of those reported for endogenous CTS in the blood, they stimulate hypertrophic growth of cultured cardiac myocytes through initiation of a Na/K-ATPase-mediated and reactive oxygen species (ROS)-dependent signaling. To examine a possible effect of endogenous concentrations of CTS on cardiac structure and function in vivo, we compared mice expressing the naturally resistant Na/K-ATPase α1 and age-matched mice genetically engineered to express a mutated Na/K-ATPase α1 with high affinity for CTS. In this model, total cardiac Na/K-ATPase activity, α1, α2, and β1 protein content remained unchanged, and the cardiac Na/K-ATPase dose–response curve to ouabain shifted to the left as expected. In males aged 3–6 months, increased α1 sensitivity to CTS resulted in a significant increase in cardiac carbonylated protein content, suggesting that ROS production was elevated. A moderate but significant increase of about 15% of the heart-weight-to-tibia-length ratio accompanied by an increase in the myocyte cross-sectional area was detected. Echocardiographic analyses did not reveal any change in cardiac function, and there was no fibrosis or re-expression of the fetal gene program. RNA sequencing analysis indicated that pathways related to energy metabolism were upregulated, while those related to extracellular matrix organization were downregulated. Consistent with a functional role of the latter, an angiotensin-II challenge that triggered fibrosis in the α1^r/rα2^s/s mouse failed to do so in the α1^s/sα2^s/s. Taken together, these results are indicative of a link between circulating CTS, Na/K-ATPase α1, ROS, and physiological cardiac hypertrophy in mice under baseline laboratory conditions.     

Mixing of a self-assembled supramolecular polymer and a covalent polymer in organic solutions

We report on a neutron scattering study of the molecular structure of a self-assembled supramolecular polymer composed of a bicopper complex within a solution of a covalent polymer (atactic polystyrene/trans-decalin). The study is achieved by putting two binary phases in contact and allowing for diffusion of both components in either phase. It is shown that the one-dimensional bicopper filaments are compatible to a high extent with the atactic polymer. These results are discussed in the light of a recently devised encapsulation process of the bicopper filaments within the fibrils of a physical network of isotactic polystyrene.