AFM imaging of immobilized fibronectin: does the surface conjugation scheme affect the protein orientation/conformation?

Covalent grafting of biomolecules could potentially improve the biocompatibility of materials. However, these molecules have to be grafted in an active conformation to play their biological roles. The present work aims at verifying if the surface conjugation scheme of fibronectin (FN) affects the protein orientation/conformation and activity. FN was grafted onto plasma-treated fused silica using two different crosslinkers, glutaric anhydride (GA) or sulfosuccinimidyl 4-(p-maleimidophenyl)butyrate (SMPB). Fused silica was chosen as a model surface material because it presents a roughness well below the dimensions of FN, therefore allowing AFM analyses with appropriate depth resolution. Cell adhesion assays were performed to evaluate the bioactivity of grafted FN. Cell adhesion was found to be higher on GA-FN than on SMPB-FN. Since FN-radiolabeling assays allowed us to rule out a surface concentration effect (approximately 80 ng/cm2 of FN on both crosslinkers), it was hypothesized that FN adopted a more active conformation when grafted via GA. In this context, the FN conformation on both crosslinkers was investigated through AFM and contact angle analyses. Before FN grafting, GA- and SMPB-modified surfaces had a similar water contact angle, topography, and roughness. However, water contact angles of GA-FN and SMPB-FN surfaces clearly show differences in surface hydrophilicity, therefore indicating a dependence of protein organization toward the conjugation strategy. Furthermore, AFM results demonstrated that surface topography and roughness of both FN-conjugated surfaces were significantly different. Distribution analysis of FN height and diameter confirmed this observation as the protein dimensions were significantly larger on GA than SMPB. This study confirmed that the covalent immobilization scheme of biomolecules influences their conformation and, hence, their activity. Consequently, selecting the appropriate conjugation strategy is of paramount importance in retaining molecule bioactivity.     

Methods to Analyse the Texture of Alginate Aerogel Microspheres

Nitrogen adsorption at 77 K has been applied to the study of the texture of alginate aerogel microspheres obtained by CO₂ supercritical drying of alcogels. The limited volume shrinkage suggests that the aerogels preserve the texture of the hydrogels. Alginate aerogels presents a N₂ adsorption at small pressure higher than reference non-porous silica, to be attributed to the polarity of the surface or to a small microporous volume. The aggregated nanobead strings of the guluronic-rich gels accounts for a significant mesoporosity. The N₂ adsorption results correspond to electron microscopy observations for features smaller than 50 nm.     

Synthesis of Polystyrene‐block‐poly(butyl acrylate) Copolymers Using Nitroxide‐Mediated Living Radical Polymerization in Miniemulsion

Living free‐radical butyl acrylate polymerization in miniemulsion was initiated by polystyrene bearing a nitroxyl end group to yield polystyrene‐block‐poly(butyl acrylate) block copolymers. Polystyrene macroinitiator was obtained using different initiating systems (potassium persulfate or benzoyl peroxide) in the presence of 2,2,6,6‐tetramethylpiperidine‐N‐oxyl (TEMPO) or the more water‐soluble 4‐hydroxy‐2,2,6,6‐tetramethylpiperidin‐N‐oxyl (OH‐TEMPO). The nitroxide water‐solubility has an important influence in determining molecular weight distribution and controlling the growth of the second block.     

DNS and LES of spark ignition with an automotive coil

The cycle to cycle combustion variability which is observed in spark-ignition engines is often caused by fluctuations of the early flame development. LES can be exploited for a better understanding and mastering of their origins. For that purpose appropriate models taking into account energy deposition, mixture ignition and transition to propagation are necessary requirements. This paper presents first DNS and LES of spark ignition with a real automotive coil and simplified pin-pin electrodes. The electrical circuit characteristics are provided by ISSIM while the energy deposition is modelled by Lagrangian particles. The ignition model is first evaluated in terms of initial spark radius on a pin-pin ignition experiment in pure air performed at CORIA and EM2C laboratories, showing that it pilots the radius of the torus formed by the initial shock wave. DNS of a quiescent lean propane/air mixture are then performed with this ignition system and a two-step mechanism. The impact of the modelled transferred energy during glow phase as well as the initial arc radius on the minimum ignition energy (MIE) are examined and compared to experimental values. Replacing the two-step chemistry by an analytically reduced mechanism leads to similar MIE but shows a different ignition kernel shape. Finally, LES of turbulent ignition using a Lagrangian arc model show a realistic prediction of the arc shape and its important role on the energy transfer location and thus on the flame kernel shape.     

Synthesis of a Phosphonate‐Linked Aminoglycoside–Coenzyme A Bisubstrate and Use in Mechanistic Studies of an Enzyme Involved in Aminoglycoside Resistance

Just five steps! The synthesis of a phosphonate‐linked aminoglycoside‐coenzyme A derivative (see scheme) that includes a Michael addition in water has been realized in just five steps.

     

Dendron‐Functionalized Core–Shell Superparamagnetic Nanoparticles: Magnetically Recoverable and Reusable Catalysts for Suzuki C?C Cross‐Coupling Reactions

A metallodendron functionalized with dicyclohexyldiphosphino palladium complex was synthesized. The metallodendron was grafted onto core–shell superparamagnetic nanoparticles (γ‐Fe₂O₃/polymer, 200–500 nm) to give optimal catalytic reactivity in cross‐coupling reactions. The grafted nanoparticles were used as recoverable and reusable catalysts for Suzuki C? C cross‐coupling reactions. They showed remarkable reactivity towards iodo‐ and bromoarenes under mild conditions, and unprecedented reactivity towards chloroarenes. On completion of the catalytic reaction, the catalysts were readily recovered by using a simple magnet to attract the superparamagnetic grafted nanoparticles. Catalysts were recovered more than 25 times with almost no discernable loss of reactivity.     

On mathematical problems as historically determined artifacts: Reflections inspired by sources from ancient China

Is a mathematical problem a cultural invariant, which would invariably give rise to the same practices, independent of the social groups considered? This paper discusses evidence found in the oldest Chinese mathematical text handed down by the written tradition, the canonical work The Nine Chapters on Mathematical Procedures and its commentaries, to answer this question in the negative. The Canon and its commentaries bear witness to the fact that, in the tradition for which they provide evidence, mathematical problems not only were questions to be solved, but also played a key part in conducting proofs of the correctness of algorithms.     

Improved method for isotopic and quantitative analysis of dissolved inorganic carbon in natural water samples

We present here an improved and reliable method for measuring the concentration of dissolved inorganic carbon (DIC) and its isotope composition (delta(13)C(DIC)) in natural water samples. Our apparatus, a gas chromatograph coupled to an isotope ratio mass spectrometer (GCIRMS), runs in a quasi-automated mode and is able to analyze about 50 water samples per day. The whole procedure (sample preparation, CO(2(g))-CO(2(aq)) equilibration time and GCIRMS analysis) requires 2 days. It consists of injecting an aliquot of water into a H(3)PO(4)-loaded and He-flushed 12 mL glass tube. The H(3)PO(4) reacts with the water and converts the DIC into aqueous and gaseous CO(2). After a CO(2(g))-CO(2(aq)) equilibration time of between 15 and 24 h, a portion of the headspace gas (mainly CO(2)+He) is introduced into the GCIRMS, to measure the carbon isotope ratio of the released CO(2(g)), from which the delta(13)C(DIC) is determined via a calibration procedure. For standard solutions with DIC concentrations ranging from 1 to 25 mmol . L(-1) and solution volume of 1 mL (high DIC concentration samples) or 5 mL (low DIC concentration samples), delta(13)C(DIC) values are determined with a precision (1sigma) better than 0.1 per thousand. Compared with previously published headspace equilibration methods, the major improvement presented here is the development of a calibration procedure which takes the carbon isotope fractionation associated with the CO(2(g))-CO(2(aq)) partition into account: the set of standard solutions and samples has to be prepared and analyzed with the same 'gas/liquid' and 'H(3)PO(4)/water' volume ratios. A set of natural water samples (lake, river and hydrothermal springs) was analyzed to demonstrate the utility of this new method.     

EXAFS and IR structural study of platinum-based anticancer drugs' degradation by diethyl dithiocarbamate

Platinum compounds constitute a discrete class of DNA-damaging anticancer drug agents, including cisplatin, carboplatin, and oxaliplatin. The toxicity of such drugs raises the problem of waste detoxification. Diethyl dithiocarbamate (DDTC) is recommended by the World Heath Organization (WHO) for the destruction of cisplatin, but the degradation product has not been structurally characterized. This paper deals with the extended X-ray absorption fine structure (EXAFS) and IR structural study of the reaction products of DDTC with cisplatin, carboplatin, and oxaliplatin. Cisplatin and carboplatin give the same reaction product: Pt(DDTC)2. In the case of oxaliplatin, we observed the formation of [(diaminocyclohexane)(DDTC)Pt(II)]. In all cases, the replacement of labile ligands by strong ligands should lead to inactive compounds. Our results suggest that the WHO inactivation protocol might be extended to carboplatin and oxaliplatin. Nevertheless, this should be validated by toxicity tests of the degradation products.