Comptes rendus de l'Academie des sciences. Serie III, Sciences de la vie最新文献

The main results published on the production of reactive oxygen intermediates by hemocytes and digestive glands of marine bivalves such as mussels, oysters or clams have been reviewed and discussed. Mussel and oyster hemocytes respond to appropriate stimuli with a burst of respiratory activity and the generation of reactive oxygen intermediates in a manner resembling the respiratory burst of mammalian phagocytes. However, interspecies differences in hemocytes-mediated antimicrobial defense mechanisms occur since clam hemocytes do not show any increase of reactive oxygen intermediate production upon similar stimulations. Hepatopancreatic gland of bivalves, as mammalian and fish liver produce reactive oxygen species during the one-electron reduction of xenobiotics, and mechanistic differences appear between bivalves and mammals. Thus, it appears that, in spite of some interspecies differences, the generation of cytotoxic reactive oxygen species is a general protective mechanism of most, if not all, animal species.

Patients with human African trypanosomiasis present a major dysruption of the circadian rhythmicity of the sleep-wake cycle, which was also found in rats infected with Trypanosoma brucei brucei (T.b.b.). The alterations in the immune function and nervous system in African trypsanosomiasis led us to investigate the involvement of nitric oxide (NO), a key molecule in immune and neurophysiological mechanisms, in experimental trypanosomiasis. NO was measured in 35 Sprague Dawley rats using differential impulsional voltammetry with a carbon fiber coated with porphyrin-nickel and nafion, ex vivo in the blood and in vivo in the brain. The rats were anaesthetized with sodium chlorate. Infection was performed intraperitoneally (i.p.) with 0.2 ml of a T.b.b. cryostabilate (clone AnTat 1.1E). Blood was collected by an intracardiac puncture with immediate replacement of blood volume (1 ml) in 7 control rats and 8 rats infected since 15 days, before and after i.p. administration of L-ANA (L-arginine-p-nitro-anilide, 100 mg.kg-1, an inhibitor of NO synthase). Brain measures were done in 20 rats (8 controls, and 12 rats infected since 15 or 21 days), in the cortex (H, -0.5 mm; AP, -0.8 mm; L, 1.2 mm) and the lateral ventricle (H,-3.2 mm). In infected rats, blood NO was at 70% of control values (p < 0.001), and L-ANA suppressed the NO signal in all animals (p < 0.0001), demonstrating that the signal originated from NO. Cortical NO was higher than in the ventricle in both control (p < 0.0001) and infected rats (p < 0.001). NO was more elevated in both structures in 15-day-infected rats than in control rats (p < 0.0001), the difference being enhanced in 21-day-infected rats (p < 0.001). L-ANA suppressed the NO signal in 30 to 60 min. These data suggest that NO intervenes in the development of trypanosomiasis in different manners. It is increased in the brain, which remains unexplained, where it may be involved in blood-brain barrier permeation. Conversely, it is decreased in the blood, may be because of macrophage function impairment, which would explain why trypanosomes can multiply in the host.

The acceptance of a pregnant female by the dominant male of a family group of alpine marmots (Marmota marmota) (population of La Grande Sassière, Parc national de la Vanoise, French Alps) was revealed by the combined results from microsatellite polymorphism analysis and behavioural studies. These first results seem to indicate that the mating system of the alpine marmot is more complex than previously thought, that polygyny cannot be excluded, and that adult females can join neighbouring groups. This acceptance would have been interpreted as an extra-pair fertilization if complete field data had not been available.

Pseudomonas aeruginosa is an opportunistic pathogen causing severe infections, especially in lungs of patients with cystic fibrosis. Environmental conditions induce the production by the bacteria of a viscous mucoid exopolysaccharide, called alginate, which is one of the most important factor of virulence of P. aeruginosa. Alginate is a linear polymer of beta-1, 4-linked L-guluronic acid and D-mannuronic acid. The alginate biosynthetic pathway involves genes called alg which are clustered at the 34 min region of chromosomal DNA of P. aeruginosa. The key enzyme of alginate biosynthesis, the GDP-mannose dehydrogenase is encoded by the gene algD. Expression of algD is positively controlled by several proteins, especially AlgU, a putative sigma factor homologous to sigma E of E. coli, AlgR and AlgP, a transactivator and an histone-like respectively. Here, a scheme of alginate biosynthetic pathway and a model for the alg genes regulation are described from results published in literature and from our own interpretation.

Seryl t-RNA synthetase of the bacterium Thermus thermophilus contains a long arm, consisting of an antiparallel coiled coil, that is involved in binding of tRNA. Two crystallographic structures exist for this protein, in which the arm is in different conformations. Here, we use computational methods employing an empirical potential energy function to investigate the flexibility of the long arm. A conformational pathway is calculated between the 2 crystallographic structures using a method based on molecular dynamics simulation. The pathway is analyzed in terms of sequential phi and psi backbone angle changes. Several transient phi and psi displacements are present along the pathway that are not visible in the end states and may be required for transition between them. Energy maps are constructed by rotating the arm around its principal axes of inertia and energy minimizing. The map identifies 2 regions of relatively low energy which might be accessible to the arm.

The repair of UV-C (254 nm) DNA lesions by nucleotide excision repair (NER) has been studied in the rodent cell line xrs6 belonging to complementation group 5 of ionising radiation sensitive (IRs) mutants. xrs6 cell line shows a defect in the DNA-end binding protein complex Ku which is involved in the repair of double-strand breaks (DSB) due to IR. In agreement with IR sensitivity, a bleomycin sensitive phenotype of xrs6 cell line was found as compared to the parental CHO-K1 line (factor > 8 fold). xrs6 exhibited also a slight (factor 2) but reproducible sensitivity to UV-C-light, while a revertant cell line for Ku DNA-end binding activity, xrs6rev, showed a restoration of both IR and UV-C sensitivities to the parental level. The NER activity of these cell lines was measured in vitro in nuclear protein extracts in the presence of plasmid DNA repair substrate damaged with UV-C lesions repaired by NER: xrs6 cell extracts exhibited only 55% of NER activity as compared to the control CHO-K1 and xrs6rev cell extracts. These results indicate that the Ku DSB repair protein is involved also in the NER process.

The development of "less invasive surgery" using small incisions and videoscopic techniques may change, in the near future, our attitude towards "traditional" surgery. This is because of specific advantages such as skin limited incisions, reduced perioperative disability and lower cost. Up to now, only abdominal, thoracic and coronary artery surgery which do not imply the opening of the heart, or closure of simple atrial septal defects, have benefitted from this new approach. This article reports the first case of open heart surgery for complex lesions of the left heart through a minithoracotomy (5 x 4 cm) with the use of videotransmission and peripheral extracorporeal circulation. The patient, a 30-year-old female, was operated upon for a combined mitral valve stenosis and insufficiency of rheumatic origin unsuccessfully treated by a previous percutaneous valve dilatation. The 2.5 h open heart procedure comprised commissurotomy, repair of torn leaflets, chordal transposition and Carpentier-Edwards prosthetic ring implantation. The patient left the hospital 12 days after the operation. Transesophageal echocardiography at discharge showed normal valve function with no residual stenosis or residual leak.

The Kin28 protein kinase interacts physically and genetically with cyclin Ccl1. Kin28 has been reported recently to be involved in the in vivo phosphorylation of the largest subunit of RNA polymerase II (Rpb1) in Saccharomyces cerevisiae. Now, we show that in a strain harboring a conditional ccl1-ts mutation, the C-terminal domain (CTD) of the Rpb1 subunit is under-phosphorylated at restrictive temperature. The transcription of a set of genes, chosen at random, is severely affected in a kin28-ts mutant shifted at restrictive temperature. Here, we report that the same set of genes requires a functional CCL1 gene product to be transcribed. These findings, added to previously published data, establishes that Kin28p is a cyclin-dependent kinase (CDK) with Ccl1p as a companion, both of them being necessary for general transcription and CTD phosphorylation.

Nitric oxide (NO) is synthesized in the neurons by constitutive NO synthase (NOS). Within given neuronal sets, this enzyme is colocalized with different other neurotransmitters such as, for example, GABA, acethylcholine or serotonin. Our attention has been focused on the fact that serotoninergic neurons, well known for their involvement in sleep triggering and maintenance, synthesize also NO. In order to evaluate the modalities of release of this compound throughout the rat sleep-waking cycle, we prepared a sensor allowing its specific detection in freely moving animals. The active part of this sensor is a carbon fiber (phi = 30 microns) successively coated with porphyrin nickel and nafion. In vitro, together with differential normal pulse voltammetric measurements, it allows the detection of a 650 mV signal varying linearly in NO solutions ranging from 5.10(-7) to 10(-4) M. At physiological concentrations, L-arginine, L-citrulline, nitrites and nitrates do not yield a signal at 650 mV. Similarly, the compounds administered to the animals, hydroxylamine, L-arginine p-nitroanilide (L-ANA) and L-N omega-nitro arginine methyl ester (L-NAME) are not electroactive at 650 mV. L-ANA and L-NAME, also appear to be trapping agents for NO while leaving the electrochemical properties of the sensor untouched. In vivo, in the frontal cortex of the anesthetized rat, a signal is measured at 650 mV. The administration of hydroxylamine (40 mg/kg, i.p.) induces a 100% increase in its height. The administration of L-ANA (100 mg/kg, i.p.) produces its complete disappearance within 50 min. Finally, the administration of L-NAME (100 mg/kg, i.p.) is without effect. This last aspect might be dependent upon the inability of L-NAME to cross the blood brain barrier. On the contrary, the increase in the signal height obtained with hydroxylamine and its disappearance with L-ANA support that it might depend upon NO. In vivo, and in animals also equipped with polygraphic electrodes, the signal measured in the same area of the cortex exhibits the highest height during the waking state and decreases during either slow-wave sleep (-6%) or paradoxical sleep (-9%). These mild variations might represent the mean of several NO sources (cortical GABAergic interneurons, cholinergic and serotoninergic axonal nerve endings), each of them varying differently throughout the sleep-waking cycle.

Isolation of homologous ras-related genes from sugar beet has never been reported. Screening cDNA library from Beta vulgaris L. Hilma resulted in the isolation of 3 ras-homologous clones. Two of these genes (rab1Bv and rab2Bv) belong to the Rab/Ypt group. The deduced polypeptidic sequences from them show strong homologies to Ara3 (93% with Rab1Bv) of Arabidopsis thaliana and Rgp2 (97% with Rab2Bv) of Oryza sativa. The third gene (rho1Bv) belongs to the Rho family. The homology of Rho1Bv protein with Rho1Ps from Pisum is very high (98%). Rho1Bv is the second representative plant Rho protein described in the literature. The homologies of all these 3 small GTP-binding proteins indicate that these proteins are conserved in plant families like Chenopodiaceae, Brassicaceae, Fabaceae and Poaceae and could control important transductional pathways conserved along the processes of evolution.