Thermogalvanoscope

Pas de Résumé disponibl

Fluorogenic detection of viable

In order to easily assess growth and destruction of Toxoplasma gondii in vitro, this report describes two double staining assays that both visualize live and dead organisms: acridine orange - ethidium bromide (AO-EB) and bisbenzimide (Hoechst 33258) - propidium iodide (B-PI). EB and PI were chosen for dead organisms staining while AO and B stain viable organisms. Thus, both double staining assays seem more informative than Giemsa staining or indirect immunofluorescence. They offer methods to study internal structure of the parasite as well as information on host-parasite relationships. Moreover, detection in culture are sensitive, easier, and less time consuming than previous methods. So, they should to be useful in strains behaviour analysis

Summer Mistral at the Exit of the Rhone Valley

The paper examines the three-dimensional structure and dynamics of the mistral at the Rhône valley exit on 28 June 2001. The mistral refers to a severe wind that develops along the Rhône valley in southern France. This summer mistral event was documented in the framework of the ESCOMPTE field experiment. The dynamical processes driving the circulation of the mistral in the Rhône valley and particularly wake formation and planetary boundary layer (PBL) inhomogeneity at the scale of Rhône valley delta are investigated. Several important data sources are used (airborne Doppler lidar, radiosondes and surface stations) as well as non-hydrostatic mesoscale simulations. This paper analyses experimentally, numerically and theoretically the mechanism of wake formation. It shows that the flow impinging on the Alpine range and the Massif Central becomes supercritical all along the ridge line, including the Rhône valley and continues to accelerate in the lee regions until a hydraulic jump occurs. It leads to the formation of wakes behind and close to the mountain peaks. Compared to the Massif Central wake, the origin of the western Alps wake is rather complicated. In this study, the observations and simulations suggest a combined wall separation/gravity wave breaking mechanism to explain the western Alps wake. Indeed, it is shown that in addition to the flow descending the western Alps slopes and experiencing a strong hydraulic jump, the point where the mistral flow separates from the eastern flank of the Rhône valley located at about 44°N is associated with a 'flank-shock' which is an oblique hydraulic jump (i.e.the downstream Froude number is supercritical). Wake formation in the lee of the Alps and the Massif Central causes large inhomogeneity of the PBL with differences between land and sea. In the Massif Central and western Alps wakes, the continental PBL is deeper (1.8 km) than in the mistral flow (1 km), which is consistent with a subcritical regime associated with enhanced turbulent mixing. The supercritical air flow, descending the Massif Central and Alps slopes and transitioning to subcritical flow, increases the near-surface air temperature due to the föhn effect. Over the Mediterranean, the surface heat fluxes are slightly negative (between–50 and 0 W m –2) and the main source of PBL turbulence is mechanical (wind shear). The PBL depth within the mistral flow does not vary over land (1 km), whereas the absence of convection but also of strong winds prevent PBL development over the sea in the wakes of the Massif Central and the Alps (PBL depth of about 0.5 km)

POLG exon 22 skipping induced by different mechanisms in two unrelated cases of Alpers syndrome.

International audienceThe POLG genes were sequenced in two unrelated patients presenting with Alpers syndrome. The novel c.3626_3629dupGATA and the c.3643+2T>C alleles were associated in trans with p.A467T and p.[W748S;E1143G], respectively. POLG transcripts from skin fibroblasts showed complete exon 22 skipping for patient 2, but surprisingly partial exon 22 skipping from the c.3626_3629dupGATA for patient 1. The creation of a putative exonic splicing silencer could be responsible for the splicing anomaly observed in patient 1. Both c.3643+2T>C and c.3626_3629dupGATA create a premature termination codon and a low polymerase γ activity in skin fibroblasts is responsible for the severe phenotype in these patients

A Major Determinant for Binding and Aminoacylation of tRNAAla in Cytoplasmic Alanyl-tRNA Synthetase Is Mutated in Dominant Axonal Charcot-Marie-Tooth Disease

Charcot-Marie-Tooth disease (CMT) is the most common cause of inherited peripheral neuropathy, with an estimated frequency of 1/2500. We studied a large family with 17 patients affected by the axonal form of CMT (CMT2). Analysis of the 15 genes or loci known to date was negative. Genome-wide genotyping identified a CMT2 locus in 16q21-q23 between D16S3050 and D16S3106. The maximum two-point LOD score was 4.77 at θ = 0 for marker D16S3050. Sequencing of candidate genes identified a unique mutation, c.986G>A (p.Arg329His), affecting a totally conserved amino acid in the helical domain of cytoplasmic alanyl-tRNA synthetase (AlaRS). A second family with the same mutation and a different founder was then identified in a cohort of 91 CMT2 families. Although mislocation of mutant Arg329His-AlaRS in axons remains to be evaluated, experimental data point mostly to a quantitative reduction in tRNAAla aminoacylation. Aminoacylation and editing functions closely cooperate in AlaRS, and Arg329His mutation could also lead to qualitative errors participating in neurodegeneration. Our report documents in 18 patients the deleterious impact of a mutation in human cytoplasmic AlaRS and broadens the spectrum of defects found in tRNA synthetases. Patients present with sensory-motor distal degeneration secondary to predominant axonal neuropathy, slight demyelination, and no atypical or additional CNS features

Kinetic properties of mutant deoxyguanosine kinase in a case of reversible hepatic mtDNA depletion

DGUOK [dG (deoxyguanosine) kinase] is one of the two mitochondrial deoxynucleoside salvage pathway enzymes involved in precursor synthesis for mtDNA (mitochondrial DNA) replication. DGUOK is responsible for the initial rate-limiting phosphorylation of the purine deoxynucleosides, using a nucleoside triphosphate as phosphate donor. Mutations in the DGUOK gene are associated with the hepato-specific and hepatocerebral forms of MDS (mtDNA depletion syndrome). We identified two missense mutations (N46S and L266R) in the DGUOK gene of a previously reported child, now 10 years old, who presented with an unusual revertant phenotype of liver MDS. The kinetic properties of normal and mutant DGUOK were studied in mitochondrial preparations from cultured skin fibroblasts, using an optimized methodology. The N46S/L266R DGUOK showed 14 and 10% residual activity as compared with controls with dG and deoxyadenosine as phosphate acceptors respectively. Similar apparent negative co-operativity in the binding of the phosphate acceptors to the wild-type enzyme was found for the mutant. In contrast, abnormal bimodal kinetics were shown with ATP as the phosphate donor, suggesting an impairment of the ATP binding mode at the phosphate donor site. No kinetic behaviours were found for two other patients with splicing defects or premature stop codon. The present study represents the first characterization of the enzymatic kinetic properties of normal and mutant DGUOK in organello and our optimized protocol allowed us to demonstrate a residual activity in skin fibroblast mitochondria from a patient with a revertant phenotype of MDS. The residual DGUOK activity may play a crucial role in the phenotype reversal