A phosphorothioate oligonucleotide blocks reverse transcription via an antisense mechanism

AbstractWe have studied the inhibition by a phosphorothioate oligodeoxynucleotide (17PScap) of cDNA synthesis performed by either avian or murine reverse transcriptase. Three different mechanisms of inhibition were identified: at low concentrations (⊃100 nM), the cleavage of the RNA template by the retroviral RNase H at the level of the RNA/17PScap duplex accounted for most of the effect, whereas hybrid-arrested cDNA synthesis by an RNase H-independent mechanism marginally contributed to the inhibition. Both mechanisms were sequence-specific. Above 100 nM, the overall cDNA synthesis was reduced in a non-specific manner

Influence of microbial community composition and metabolism on air.sea ΔpCO 2 variation off the western Antarctic Peninsula

We studied CO 2 and O 2 dynamics in the western Antarctic Peninsula (WAP) waters in relation to (1) phytoplankton biomass, (2) microbial community primary production and respiration, and (3), for the first time, phytoplankton composition, during summer and fall in 3 consecutive years (2002, 2003 and 2004). The areal average of ΔpCO 2 (the difference between surface seawater and atmospheric partial pressure of CO 2) for the 3 yr was significantly negative (.20.04 ± 44.3 μatm, p < 0.01) during the summer to fall period in the region, possibly indicating a CO 2 sink. In the southern WAP (i.e. south of Anvers Island), ΔpCO 2 was significantly negative (.43.60 ± 39.06 μatm) during fall. In the northern WAP (north of Anvers Island), ΔpCO 2 values showed a more complex distribution during summer and fall (.4.96 ± 37.6 and 21.71 ± 22.39 μatm, respectively). Chlorophyll a (chl a) concentration averaged 1.03 ± 0.25 μg l.1 and was higher in the south of the peninsula. Phytoplankton composition influenced chl a concentration with higher and lower values for diatom-and phytoflagellate-dominated communities, respectively. A significant negative correlation existed between chl a and ΔpCO 2. From incubation experiments performed in the northern WAP, respiration was low (averaging 5.1 mmol O 2 m.3 d.1), and the net community production (NCP) correlated negatively with ΔpCO 2 and positively with %O 2 saturation. However, despite the high NCP values measured, ΔpCO 2 was significantly positive in the northern WAP during the summer to fall period. Strong mixing and lower chl a concentration may explain this result. In contrast, ΔpCO 2 was significantly negative in the southern WAP, possibly because of high surface water chl a concentration.Facultad de Ciencias Naturales y Muse

Zip Nucleic Acids: new high affinity oligonucleotides as potent primers for PCR and reverse transcription

Most nucleic acid-based technologies rely upon sequence recognition between an oligonucleotide and its nucleic acid target. With the aim of improving hybridization by decreasing electrostatic repulsions between the negatively charged strands, novel modified oligonucleotides named Zip nucleic acids (ZNAs) were recently developed. ZNAs are oligonucleotide–oligocation conjugates whose global charge is modulated by the number of cationic spermine moieties grafted on the oligonucleotide. It was demonstrated that the melting temperature of a hybridized ZNA is easily predictable and increases linearly with the length of the oligocation. Furthermore, ZNAs retain the ability to discriminate between a perfect match and a single base-pair-mismatched complementary sequence. Using quantitative PCR, we show here that ZNAs are specific and efficient primers displaying an outstanding affinity toward their genomic target. ZNAs are particularly efficient at low magnesium concentration, low primer concentrations and high annealing temperatures, allowing to improve the amplification in AT-rich sequences and potentially multiplex PCR applications. In reverse transcription experiments, ZNA gene-specific primers improve the yield of cDNA synthesis, thus increasing the accuracy of detection, especially for genes expressed at low levels. Our data suggest that ZNAs exhibit faster binding kinetics than standard and locked nucleic acid-containing primers, which could explain why their target recognition is better for rare targets

CFD for the Study of the Multiphase Flow and Mass Transfer in a Stirred Tank Reactor for the Screening of Shaped Catalysts

International audienc

Enjeux théoriques et pratiques du développement régional : 30 Ans de recherche au GRIR

Le trentième anniversaire du GRIR a été souligné au printemps 2013 par un colloque regroupant plusieurs chercheurs dans le domaine du développement local et régional. Cet ouvrage est le produit de leurs réflexions tant sur le plan théorique que pratique. Ce livre est articulé autour de trois axes : 1) la présentation des divers courants en développement régional, 2) des réflexions sur le passé et le présent des Premières Nations et 3) la présentation de cas concrets sur la prise en main du développement par les différents acteurs du territoire. Cet ouvrage est un outil permettant de constater le travail accompli en développement local et régional et de dégager des pistes de réflexion sur les avenues possibles de développement

Influence of microbial community composition and metabolism on air.sea ΔpCO 2 variation off the western Antarctic Peninsula

We studied CO 2 and O 2 dynamics in the western Antarctic Peninsula (WAP) waters in relation to (1) phytoplankton biomass, (2) microbial community primary production and respiration, and (3), for the first time, phytoplankton composition, during summer and fall in 3 consecutive years (2002, 2003 and 2004). The areal average of ΔpCO 2 (the difference between surface seawater and atmospheric partial pressure of CO 2) for the 3 yr was significantly negative (.20.04 ± 44.3 μatm, p < 0.01) during the summer to fall period in the region, possibly indicating a CO 2 sink. In the southern WAP (i.e. south of Anvers Island), ΔpCO 2 was significantly negative (.43.60 ± 39.06 μatm) during fall. In the northern WAP (north of Anvers Island), ΔpCO 2 values showed a more complex distribution during summer and fall (.4.96 ± 37.6 and 21.71 ± 22.39 μatm, respectively). Chlorophyll a (chl a) concentration averaged 1.03 ± 0.25 μg l.1 and was higher in the south of the peninsula. Phytoplankton composition influenced chl a concentration with higher and lower values for diatom-and phytoflagellate-dominated communities, respectively. A significant negative correlation existed between chl a and ΔpCO 2. From incubation experiments performed in the northern WAP, respiration was low (averaging 5.1 mmol O 2 m.3 d.1), and the net community production (NCP) correlated negatively with ΔpCO 2 and positively with %O 2 saturation. However, despite the high NCP values measured, ΔpCO 2 was significantly positive in the northern WAP during the summer to fall period. Strong mixing and lower chl a concentration may explain this result. In contrast, ΔpCO 2 was significantly negative in the southern WAP, possibly because of high surface water chl a concentration.Facultad de Ciencias Naturales y Muse

Natural mutations in the sensor kinase of the PhoPR two-component regulatory system modulate virulence of ancestor-like tuberculosis bacilli

The molecular factors and genetic adaptations that contributed to the emergence of Mycobacterium tuberculosis (MTB) from an environmental Mycobacterium canettii-like ancestor, remain poorly investigated. In MTB, the PhoPR two-component regulatory system controls production and secretion of proteins and lipid virulence effectors. Here, we describe that several mutations, present in phoR of M. canettii relative to MTB, impact the expression of the PhoP regulon and the pathogenicity of the strains. First, we establish a molecular model of PhoR and show that some substitutions found in PhoR of M. canettii are likely to impact the structure and activity of this protein. Second, we show that STB-K, the most attenuated available M. canettii strain, displays lower expression of PhoP-induced genes than MTB. Third, we demonstrate that genetic swapping of the phoPR allele from STB-K with the ortholog from MTB H37Rv enhances expression of PhoP-controlled functions and the capacities of the recombinant strain to colonize human macrophages, the MTB target cells, as well as to cause disease in several mouse infection models. Fourth, we extended these observations to other M. canettii strains and confirm that PhoP-controlled functions are expressed at lower levels in most M. canettii strains than in M. tuberculosis. Our findings suggest that distinct PhoR variants have been selected during the evolution of tuberculosis bacilli, contributing to higher pathogenicity and persistence of MTB in the mammalian host

CFD for the Study of the Multiphase Flow and Mass Transfer in a Stirred Tank Reactor for the Screening of Shaped Catalysts

International audienc

Septins suppress the release of vaccinia virus from infected cells.

Septins are conserved components of the cytoskeleton that play important roles in many fundamental cellular processes including division, migration, and membrane trafficking. Septins can also inhibit bacterial infection by forming cage-like structures around pathogens such as Shigella We found that septins are recruited to vaccinia virus immediately after its fusion with the plasma membrane during viral egress. RNA interference-mediated depletion of septins increases virus release and cell-to-cell spread, as well as actin tail formation. Live cell imaging reveals that septins are displaced from the virus when it induces actin polymerization. Septin loss, however, depends on the recruitment of the SH2/SH3 adaptor Nck, but not the activity of the Arp2/3 complex. Moreover, it is the recruitment of dynamin by the third Nck SH3 domain that displaces septins from the virus in a formin-dependent fashion. Our study demonstrates that septins suppress vaccinia release by "entrapping" the virus at the plasma membrane. This antiviral effect is overcome by dynamin together with formin-mediated actin polymerization

The HIV-1 Integrase α4-Helix Involved in LTR-DNA Recognition Is also a Highly Antigenic Peptide Element

Monoclonal antibodies (MAbas) constitute remarkable tools to analyze the relationship between the structure and the function of a protein. By immunizing a mouse with a 29mer peptide (K159) formed by residues 147 to 175 of the HIV-1 integrase (IN), we obtained a monoclonal antibody (MAba4) recognizing an epitope lying in the N-terminal portion of K159 (residues 147–166 of IN). The boundaries of the epitope were determined in ELISA assays using peptide truncation and amino acid substitutions. The epitope in K159 or as a free peptide (pep-a4) was mostly a random coil in solution, while in the CCD (catalytic core domain) crystal, the homologous segment displayed an amphipathic helix structure (α4-helix) at the protein surface. Despite this conformational difference, a strong antigenic crossreactivity was observed between pep-a4 and the protein segment, as well as K156, a stabilized analogue of pep-a4 constrained into helix by seven helicogenic mutations, most of them involving hydrophobic residues. We concluded that the epitope is freely accessible to the antibody inside the protein and that its recognition by the antibody is not influenced by the conformation of its backbone and the chemistry of amino acids submitted to helicogenic mutations. In contrast, the AA →Glu mutations of the hydrophilic residues Gln148, Lys156 and Lys159, known for their interactions with LTRs (long terminal repeats) and inhibitors (