Syntheses of poly[N-(2,2 dimethoxyethyl)-N-methyl acrylamide] for the immobilization of oligonucleotides

Radical-initiated polymerization of N-(2,2 dimethoxyethyl)-N-methylacrylamide has been carried out either in chloroform or methanol using 2,2′-azobisisobutyronitrile as an initiator, allowing us to prepare acetal containing water-soluble polymers. A kinetic study in both solvents showed that this monomer fairly homopolymerized (k_p · k_t^−1/2 = 1 mol^−1/2 L^1/2 s^−1/2). Static light scattering was used to characterize the molecular weight of these polymers. In addition, the Mark–Houwink–Sakurada relationship was established based on viscosity measurements performed at 25°C in water. Recovery of the aldehyde moieties on the polymer was achieved under mild conditions using a diluted inorganic solution. The analysis of the formation of aldehyde groups was performed by ¹H- and ¹³C-NMR. The covalent binding of oligodeoxyribonucleotides was carried out in water/acetonitrile mixtures with subsequent NaBH₄ reduction of the imine bonds so as to stabilize the polymer/oligodeoxynucleotide conjugates. © 1996 John Wiley & Sons, Inc.     

Oxidation products of polyunsaturated fatty acids in infant formulas compared to human milk--a preliminary study

Information about lipid oxidation in fresh and stored human milk compared with infant formulas is scarce. We aimed to assess n-6 and n-3 PUFA oxidation in these milks by measuring the 4-hydroxynonenal (4-HNE) and 4-hydroxyhexenal (4-HHE) content. Human milk samples (n = 4), obtained from volunteer mothers, were analyzed fresh and after 1 wk at 4 degrees C or 24 h at 18 degrees C. Vitamin E and malondialdehyde (MDA) were measured by HPLC and fatty acid profile by GC. The 4-HHE and 4-HNE contents were measured by GC-MS. Infant formulas (n = 10) were tested; their fat droplet size was measured by laser light scattering and observed by confocal laser scanning microscopy. Human milk samples contained 31.0 +/- 6.3 g/L of lipids and 1.14 +/- 0.26 mg/L of vitamin E. Fat droplets were smaller in infant formulas than reported in human milk. The (4-HHE/n-3 PUFA) ratio was 0.19 +/- 0.01 microg/g in fresh human milk (unchanged after storage) versus 3.6 +/- 3.1 microg/g in dissolved powder formulas and 4.3 +/- 3.8 microg/g in liquid formula. (4-HNE/n-6 PUFA) was 0.004 +/- 0.000 microg/g in fresh milk (0.03 +/- 0.01 microg/g after storage) versus 1.1 +/- 1.0 microg/g in dissolved powder formulas and 0.2 +/- 0.3 microg/g in liquid formula. Infant formulas also contained more MDA than human milk. n-3 PUFA were more prone to oxidation than n-6 PUFA. Whether threshold levels of 4-HHE and 4-HNE would be of health concern should be elucidated.     

Involvement of GPx-3 in the Reciprocal Control of Redox Metabolism in the Leukemic Niche

The bone marrow (BM) microenvironment plays a crucial role in the development and progression of leukemia (AML). Intracellular reactive oxygen species (ROS) are involved in the regulation of the biology of leukemia-initiating cells, where the antioxidant enzyme GPx-3 could be involved as a determinant of cellular self-renewal. Little is known however about the role of the microenvironment in the control of the oxidative metabolism of AML cells. In the present study, a coculture model of BM mesenchymal stromal cells (MSCs) and AML cells (KG1a cell-line and primary BM blasts) was used to explore this metabolic pathway. MSC-contact, rather than culture with MSC-conditioned medium, decreases ROS levels and inhibits the Nrf-2 pathway through overexpression of GPx3 in AML cells. The decrease of ROS levels also inactivates p38MAPK and reduces the proliferation of AML cells. Conversely, contact with AML cells modifies MSCs in that they display an increased oxidative stress and Nrf-2 activation, together with a concomitant lowered expression of GPx-3. Altogether, these experiments suggest that a reciprocal control of oxidative metabolism is initiated by direct cell–cell contact between MSCs and AML cells. GPx-3 expression appears to play a crucial role in this cross-talk and could be involved in the regulation of leukemogenesis.     

Adhesion of edible oils to food contact surfaces

Adhesion of oils and fatty food products to packages is an important storage problem, because it increases product-package interactions that alter quality. Reducing such adhesion would also allow savings in recycling and cleaning processes. The aim of our work was to test if some thermodynamical adhesion models were correlated to edible oils’ bulk adhesion as measured experimentally. Food-contact surfaces were low-density polyethylene, polyethylene teraphthalate, stainless steel, and glass. The Young-Dupré equation and five models of adhesion from the literature were used to calculate solids’ surface tension and the thermodynamical work of adhesion (Wa). The dispersive, polar, acid-base, and hydrogen surface tension components of oils and solids were calculated. The experimental adhesion, or amount of edible oils remaining on solid surfaces after contact, was found to be correlated to Young-Dupré Wa, involving contact angle measured by specially designed image analysis technique. Two models, involving, respectively, surface tension’s hydrogen component and a linear dependence of Wa^p on the liquid polar surface tension component, fitted best with oil bulk adhesion as measured experimentally. Our theoretical approach to fatty food material adhesion seems, so far, consistent to predict global residues of edible oils on solid surfaces.     

The Oxoglutarate Binding Site and Regulatory Mechanism Are Conserved in Ammonium Transporter Inhibitors GlnKs from Methanococcales

Protein inhibition is a natural regulatory process to control cellular metabolic fluxes. P_II-family signal-transducing effectors are in this matter key regulators of the nitrogen metabolism. Their interaction with their various targets is governed by the cellular nitrogen level and the energy charge. Structural studies on GlnK, a P_II-family inhibitor of the ammonium transporters (Amt), showed that the T-loops responsible for channel obstruction are displaced upon the binding of 2-oxoglutarate, magnesium and ATP in a conserved cleft. However, GlnK from Methanocaldococcus jannaschii was shown to bind 2-oxoglutarate on the tip of its T-loop, causing a moderate disruption to GlnK–Amt interaction, raising the question if methanogenic archaea use a singular adaptive strategy. Here we show that membrane fractions of Methanothermococcus thermolithotrophicus released GlnKs only in the presence of Mg-ATP and 2-oxoglutarate. This observation led us to structurally characterize the two GlnK isoforms apo or in complex with ligands. Together, our results show that the 2-oxoglutarate binding interface is conserved in GlnKs from Methanococcales, including Methanocaldococcus jannaschii, emphasizing the importance of a free carboxy-terminal group to facilitate ligand binding and to provoke the shift of the T-loop positions.     

Development and characterization of cellulose/clay nanocomposites

Cotton is the most important textile fiber for apparel use and is preferred to synthetic fibers for reasons such as comfort and feel. A major drawback of cellulosic fibers is flammability. The development of cellulose/clay nanocomposites for use as flame-retardant materials based on cotton is reported in this paper. Novel nanocomposite materials have been produced from cellulose with layered silicate clays used as the nanofiller material. Three different methods were attempted in producing these organic–inorganic hybrids. The nanocomposites show significant improvements in thermal properties when compared with cellulose control sources. The degradation temperature of the nanocomposites increased by 45 °C and the char yields for some compositions doubled those of the controls. The crystalline melt of the materials decreased by 15 °C. Tensile testing revealed an increase of approximately 80% in the ultimate stress of the cellulose/clay nanocomposites.     

Integration of a uniform and rapid heating source into microfluidic systems

The purpose of this study has been to demonstrate the possibilities of uniform heating of a cavity, with great accuracy, by means of an integrated resistor built with the same dimensions as the cavity, i.e., with a high level of integration. For application purposes, a compact resistor allows increasing the number of cavities in which temperature can be independently controlled on the same substrate, which can prove critical for high-throughput screening applications. Potential applications lie in the field of biology or chemistry. In order to achieve the desired result, an optimization procedure was performed on the shape of the resistor. The heater size reduction enables a high level of integration with a reduced heating source surface area. Resistor shape has been optimized to reduce the influence of boundary effects, using improvements introduced in genetic algorithms. An experimental validation of the temperature profile inside the cavity has been carried out using a dye whose fluorescence depends on temperature, i.e., Rhodamine B, it will be shown herein that the optimized resistor allows for temperature cycling, e.g., for PCR applications.     

Phospholipid species and minor sterols in French human milks

The purpose of the study was to characterise the phospholipid and sterol composition in four French breast milk samples, with an aim to highlight molecular lipid species of possible nutritional significance for infants. Main PL species were phosphatidylethanolamine (PE, 21.3 ± 4.7%), phosphatidylcholine (19.0 ± 2.2%) and sphingomyelin (43.3 ± 2.6%). PE contained more arachidonic acid (4.8%) than did other PL species (P < 0.001). PE and PS + PI contained the highest proportion of DHA among PL species (0.94% and 1.13%, respectively, P < 0.05). Several minor bioactive sterols were detected in the polar lipids of human milks, e.g., desmosterol, lathosterol, lanosterol, stigmasterol and β-sitosterol. The metabolic significance and health impact of this lipid consumption by the infant should thus be explored.     

A new method to determine the microbial kinetic parameters in biological air filters

This paper presents a new method to determine kinetic parameters of the biodegradation of various pollutants in a biofilter. Toluene, a readily biodegradable volatile organic compound, and methane, a hydrocarbon and a greenhouse gas, have been chosen as the target pollutants. The new protocol utilized biomass immobilized on bed pellets; these directly sampled from a continuous steady-state biofilter. The comparison of this method with the conventional experimental protocol utilizing micro-organisms suspended in a liquid medium was made using the pollutant toluene. Indeed, with both methods, the kinetic parameters have been evaluated by following the microbial growth in batch, thermostated reactors, using determined amounts of pollutant substrate. This experiment has confirmed the pertinence of the new procedure. The interesting features of the new method are that: (1) it is easy to operate (no preliminary treatment of the bed samples) and (2) it provides reproducible parameters that represent the real biofilter case more adequately than liquid cultures. In addition, modeling of the experimental specific growth rates in the case of toluene has shown that the values obtained with the use of solid extracts can be correlated by a Haldane's formulation, where , , and . The maximum specific growth rate was reached for an initial concentration of toluene near . The determination of the experimental specific growth rates of micro-organisms in the methane biofilter has also been performed. This study allowed highlighting two methane concentrations’ ranges: from 1000 to 14 500 ppmv and from 14 500 to 27 000 ppmv. For the first range, the Monod model proves to be suitable with the kinetic parameters: and . For the second range, neither the Monod nor the Haldane's formulation could directly be used. However, a mathematical adjustment of the Monod model allows to find kinetic parameters and . The biomass yields for the tested methane concentrations have also been determined and showed two different tendencies, depending on the same two ranges. For the first range of methane concentrations, the biomass yield was quite constant with an average value around while for the second range, it could be approached by a polynomial second-order regression. The maximum value of the biomass yield obtained on the second range was at a methane initial concentration of 20 000 ppmv.