Detection of genetically modified corn (Bt176) in spiked cow blood samples by polymerase chain reaction and immunoassay methods

The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) and the 5'-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (alpha(s1)-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood.     

OH�� Radical Production Induces Direct Enhancement of the Amylo?d �� Protein/Chondroitin Sulfate Binding: Inhibition by Potentially Radical Scavengers, a Biochromatographic and Thermodynamic Model

??-Amyloid (A??) is a major component of the senile plaques characteristic of Alzheimer disease (AD). Chondroitin sulfate (CS) and glycoaminoglycan (GAG) are also localized throughout the senile plaques in AD. In previous studies, the interaction of the A?? protein with CS immobilized on a chromatographic support and the role of aluminum and copper cations was studied using a molecular chromatographic approach [1, 2]. Here, we demonstrated the direct implication of OH· radical formation on this binding via a novel analytical procedure. The binding of A?? amyloid on CS was accompanied by an OH· radical uptake. The A?¨CCS complex was stabilized by the OH· radical via the creation of about one to two hydrogen bonds. The addition in the medium of a radical scavenger allowed decreasing the A??/CS association and thus confirmed the positive role of these compounds in amyloidosis.     

The Influence of the Calcination and Water Washing Treatments on the Uranium Content in Sedimentary Phosphate Samples Using Solid State Nuclear Track Detectors and Gamma-Ray Spectrometry

The uranium and thorium contents were evaluated in the 100–400 µm granulometric fraction of different sedimentary phosphate samples by using a method based on determining the mean critical angles of etching of the CR-39 and LR-115 type II solid state nuclear track detectors (SSNTD) for detecting -particles emitted by the nuclei of the uranium-238 and thorium-232 series. Data obtained were compared with the results of -ray spectrometry measurements performed on the same samples. The influence of the calcination and water washing treatments as well as the lithology and stratigraphy on the uranium concentration of the phosphate samples was investigated.     

A HTS Assay for the Detection of Organophosphorus Nerve Agent Scavengers

A new pro‐fluorescent probe aimed at a HTS assay of scavengers is able to selectively and efficiently cleave the P? S bond of organophosphorus nerve agents and by this provides non‐toxic phosphonic acid has been designed and synthesised. The previously described pro‐fluorescent probes were based on a conventional activated P? Oaryl bond cleavage, whereas our approach uses a self‐immolative linker strategy that allows the detection of phosphonothioase activity with respect to a non‐activated P? Salkyl bond. Further, we have also developed and optimised a high‐throughput screening assay for the selection of decontaminants (chemical or biochemical scavengers) that could efficiently hydrolyse highly toxic V ‐type nerve agents. A preliminary screening, realised on a small α‐nucleophile library, allowed us to identify some preliminary “hits”, among which pyridinealdoximes, α‐oxo oximes, hydroxamic acids and, less active but more original, amidoximes were the most promising. Their selective phosphonothioase activity has been further confirmed by using PhX as the substrate, and thus they offer new perspectives for the synthesis of more potent V nerve agent scavengers.     

Homoleptic Zincate‐Promoted Room‐Temperature Halogen–Metal Exchange of Bromopyridines

Homoleptic lithium tri‐ and tetraalkyl zincates were reacted with a set of bromopyridines. Efficient and chemoselective bromine–metal exchanges were realized at room temperature with a substoichiometric amount of nBu₄ZnLi₂?TMEDA reagent (1/3 equiv; TMEDA=N,N,N′,N′‐tetramethylethylenediamine). This reactivity contrasted with that of tBu₄ZnLi₂?TMEDA, which was inefficient below one equivalent. DFT calculations allowed us to rationalize the formation of N???Li stabilized polypyridyl zincates in the reaction. The one‐pot difunctionalization of dibromopyridines was also realized using the reagent stoichiometrically. The direct creation of C? Zn bonds in bromopyridines enabled us to perform efficient Negishi‐type cross‐couplings.     

On the use of symmetry in the ab initio quantum mechanical simulation of nanotubes and related materials

Nanotubes can be characterized by a very high point symmetry, comparable or even larger than the one of the most symmetric crystalline systems (cubic, 48 point symmetry operators). For example, N = 2n rototranslation symmetry operators connect the atoms of the (n,0) nanotubes. This symmetry is fully exploited in the CRYSTAL code. As a result, ab initio quantum mechanical large basis set calculations of carbon nanotubes containing more than 150 atoms in the unit cell become very cheap, because the irreducible part of the unit cell reduces to two atoms only. The nanotube symmetry is exploited at three levels in the present implementation. First, for the automatic generation of the nanotube structure (and then of the input file for the SCF calculation) starting from a two‐dimensional structure (in the specific case, graphene). Second, the nanotube symmetry is used for the calculation of the mono‐ and bi‐electronic integrals that enter into the Fock (Kohn‐Sham) matrix definition. Only the irreducible wedge of the Fock matrix is computed, with a saving factor close to N. Finally, the symmetry is exploited for the diagonalization, where each irreducible representation is separately treated. When M atomic orbitals per carbon atom are used, the diagonalization computing time is close to Nt, where t is the time required for the diagonalization of each 2M × 2M matrix. The efficiency and accuracy of the computational scheme is documented. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010     

Molecular hydrogen "pairing" interaction in a metal organic framework system with unsaturated metal centers (MOF-74)

Infrared (IR) absorption spectroscopy measurements of molecular hydrogen in MOF-74-M (M = metal center) are performed as a function of temperature and pressure [to 45 kTorr (60 bar) at 300 K, and at lower pressures in the 20-200 K range] to investigate the nature of H(2) interactions with the unsaturated metal centers. A small shift (～?-30 cm(-1) with respect to the unperturbed H(2) molecule) is observed for the internal stretch frequency of H(2) molecules adsorbed on the metal site at low loading. This finding is in contrast to much larger shifts (～?-70 cm(-1)) observed in previous studies of MOFs with unsaturated metal centers (including MOF-74) and the general assumption that H(2) stretch shifts depend on adsorption energies (FitzGerald et al., Phys. Rev. B 2010, 81, 104305). We show that larger shifts (～?-70 cm(-1)) do occur, but only when the next available site ("oxygen" site) is occupied. This larger shift originates from H(2)-H(2) interactions on neighboring sites of the same pore, consistent with the short distance between H(2) in these two sites ～2.6 ? derived from an analysis of neutron diffraction experiments of D(2)-D(2) at 4 K (Liu et al., Langmuir 2008, 24, 4772-4777). Our results at 77 K and low loading can be explained by a diffusion barrier against pair disruption, which should be enhanced by this interaction. Calculations indicate that the vibrational shifts do not correlate with binding energies and are instead very sensitive to the environment (interaction potential and H(2)-H(2) interactions), which complicates the use of variable temperature IR methods to calculate adsorption energies of specific adsorption sites.     

Self-induced "electroclick" immobilization of a copper complex onto self-assembled monolayers on a gold electrode

We report the self-induced "electroclick" immobilization of the [Cu(II)(6-ethynyl-TMPA)(H(2)O)](2+) complex, by its simple electro-reduction, onto a mixed azidoundodecane-/decane-thiol modified gold electrode. The redox response of the grafted [Cu(II/I)(TMPA)] at the modified electrode is fully reversible indicating no Cu coordination change and a fast electron transfer.     

Phosphino-(α-sulfinylalkyl)phosphonium ylide complexes (Rh,Pd) with a configurationally stable asymmetric ylidic carbon

The synthesis and structure of Rh(I) and Pd(II) complexes of chiral P,C-chelating phosphino-(α-sulfinylalkyl)phosphonium ylide ligands with a trisubstituted asymmetric ylidic center P⁺–C1R(S1(O)p-Tol)–M (R = alkyl group) have been investigated, and compared to those of the analogous disubstituted ylide complexes (R = H). Reaction of the ethyl onium ylide of o-bis(diphenylphosphino)benzene with (?)-menthyl-(S)-p-tolylsulfinate afforded the corresponding racemic erythro phosphino-(α-sulfinylethyl)phosphonium in 90% de (R = Me). The racemization process is interpreted by a Berry-like pseudorotation mechanism driven by the steric repulsion between the α-methyl substituent and the bulky menthyloxy S-substituent or sulfur lone pair in the intermediate ylide-sulfinyl adduct. The ylide of phosphino-(α-sulfinylethyl)phosphonium reacts with [Rh(cod)₂][PF₆] and PdCl₂(MeCN)₂ to afford the corresponding P,C1-chelated threo-Rh(I) and erythro-Pd(II) mononuclear complexes in 70% yield and total diastereoselectivity. These respective complexes act as efficient catalytic precursors for the hydrogenation of (Z)-α-acetamidocinnamic acid and allylic substitution of 3-acetoxy-1,3-diphenyl-1-propene with sodium dimethyl malonate. The bonding features of the erythro-Pd(II) complex exhibiting a sulfinyl O?Pd interaction are studied theoretically at the DFT level using ELF and MESP analyses. The η²-P,C haptomeric form of the ylide ligand is estimated to compete at 19% with the η¹-C haptomeric form dominating at 81%.