Enhanced stability of carboxypeptidase from Sulfolobus solfataricus at high pressure

Carboxypeptidase from the archaebacterium Sulfolobus solfataricus is heat stable with an optimal enzyme activity at 85 °C (Colombo et al., 1992). However in the absence of glycerol and beta-mercaptoethanol, at 50 °C, the enzyme undergoes a slow thermal inactivation upon dilution in an aqueous buffer at pH 6.5. This loss of activity can be inhibited when the enzyme is maintained at high pressure. At higher temperatures, higher pressures (up to 400 MPa) are required to maintain the enzyme in its active state.Dedicated to Dr. Andrea Villa, who deceased accidentally in the course of this work.     

Furoquinoline alkaloid accumulation in Fagara zanthoxyloides cell cultures is highly dependent on the presence of exogenous benzylaminopurine

Culturing a non-habituated cell line of Fagara zanthoxyloides Lam. (Rutaceae) in either auxin-free medium or cytokinin-free medium led to opposite effects on furoquinoline accumulation. It appeared that production of skimmianine and -fagarine in the cells was strongly correlated with the presence of exogenous BAP: the levels of both alkaloids were 9 times lower when cells were cultured without cytokinin than in the control culture. NAA removal induced a slight stimulation of skimmianine and -fagarine accumulations, 1.2 and 1.9 times respectively. Culturing the cells in a PGR-free medium generated skimmianine and -fagarine levels that were 3.5 and 2.1 times lower, respectively, confirming the opposite effects of BAP and NAA on furoquinoline accumulation. Growth was only slightly inhibited when cells were cultured for one passage in the PGR-modified media.Abbreviations BAP 6-benzylaminopurine - MS Murashige-Skoog - NAA -naphtaleneacetic acid - PGR plant growth regulator     

Synthesis and conformational analysis of AzAsx-containing oligopeptides

Summary For the sake of improving synthetic methods and evaluating the conformational perturbation induced by the substitution of AzAsx for the Asx residue in the cognate dipeptide R-CO-Asx-Pro-NHR, we prepared the AzAsx-dipeptide sequence by making use of the crystalline triphosgene. This reagent allows, in situ and under very mild experimental conditions, both the carbonylation and the activation of the properly substituted and N-protected hydrazine before coupling with the proline partner. With regard to conformational behaviour, the azadipeptide sequence displays a -fold, unlike the cognate dipeptide which adopts an Asx-turn.     

Micronutrient composition of xylem sap and needles as a result of P-fertilization in maritime pine

Xylem sap and foliar compositions (P, Cu, Zn, Mn) were examined over 4 months in control and P-fertilized maritime pine in a 7-year-old plantation. The absorption of copper, zinc, and boron appears to be reduced by phosphate fertilization. Manganese concentrations are positively related to P-supply increase. Foliar analysis and sap provides the same type of information in terms of antagonism or synergism between elements but the response to treatment is more often significant with sap throughout the seasons and is completely non-existent on a single classical autumn diagnosis with foliar concentration (i.e. Zn and Cu). In consequence foliar analysis is not sensitive enough for copper or zinc deficiency diagnoses in young plantations where nutritional growth disorders appear in a short period of spring, and xylem chemistry appears to be a convenient tool in monitoring micronutrient disorders in pine plantations.     

Treatment of PC12 cells by nerve growth factor,dexamethasone, and forskolin

Several lines of anatomical, neurochemical, electrophysiological, and behavioral evidence suggest the existence of physiological interactions between neurotensin (NT) and the brain dopaminergic systems. Thus, NT has been shown to exert a neuroleptic-like action and could be implicated in the pathogenesis and treatment of schizophrenia. It is thus of particular importance to develop in vitro cell culture systems as models to study such interactions. Rat adrenal pheochromocytoma PC12 cells, which expressed high levels of tyrosine hydroxylase, were used in the present study. In contrast to rat brain cells in primary cultures, PC12 cells did not express functional NT receptors. However, they were able to express both NTmRNA and NT in response to NGF, forskolin, and dexamethasone. Those neurochemical modifications furthermore may be related to changes in the morphology of the PC12 cells in response to NGF, forskolin, and dexamethasone alone or in combination. These data suggest that PC12 cells may provide a useful model to study in vitro the regulation of both catecholamine and neurotensin phenotypes.     

Genetic analysis of new French X-linked juvenile retinoschisis kindreds using microsatellite markers closely linked to the RS locus: further narrowing of the RS candidate region

The gene involved in juvenile retinoschisis (RS) has previously been localized, by genetic linkage analyses, to Xp22.1-p22.2, between DXS274 and DXS43/ DXS207; it is closely linked to the latter markers. From our recent data, this interval represents a genetic distance of approximately 10 cM. In the present study, we have studied 14 French families with X-linked juvenile RS by using four CA polymorphisms that are closely linked to the RS locus and that have recently been included in an Xp22.1-p22.2 high-resolution map. Complete cosegregation with the disease locus was observed for three of them, DXS207, DXS418, and DXS999, which further confirms the locus homogeneity for RS and the close linkage to this region. One recombinant was found with the most proximal marker, AFM291wf5, thereby defining this marker as the new proximal boundary of the candidate region for RS. Under the assumption that DXS207 and DXS43 constitute the distal boundary, the present study further reduces the region containing the disease gene to a interval of 3–4 cM. The results reported here should facilitate the eventual cloning of the RS gene.     

Enhanced secretion through the Shigella flexneri Mxi-Spa translocon leads to assembly of extracellular proteins into macromolecular structures

Genes required for entry of Shigella flexneri into epithelial cells in vitro are clustered in two adjacent loci, one of which encodes secretory proteins, the IpaA–D proteins, and the other their dedicated secretion apparatus, the Mxi–Spa translocon. Ipa secretion, which is induced upon contact of bacteria with epithelial cells, is prevented during growth in vitro. Here, we show that ipaB and ipaD mutations lead to enhanced secretion of a set of about 15 proteins. These extracellular proteins and some Ipas associate in organized structures consisting of extended sheets. Growth of the wild-type strain in the presence of Congo red is shown to induce protein secretion through the Mxi–Spa translocon. Cultures grown to stationary phase in the presence of Congo red contain extracellular filaments whose composition and morphology are similar to those produced by the hyper-secreting ipaB and ipaD mutants.     

Effect of sulphate deficiency on the plasma membrane polypeptide composition of Brassica napus

Highly purified oilseed rape (Brassica napus) root plasma membranefractions were prepared and their polypeptide patterns analysedby two-dimensional gel electrophoresis. Sulphur starvation enhancesthe sulphate uptake capacity of B. napus roots. The relativeabundance of several polypeptides increased significantly andspecifically after sulphur starvation. Several of them (37,38, 60, and 65 kDa), found in sulphur-starved plants, were moreabundant in a phase-partitioned membrane fraction treated withTriton X-100/KBr, indicating that they are intrinsic polypeptides.One polypeptide (47 kDa) was identified in the in vitro translationproducts of the roots mRNAs as specific for S-starved plants.It was also present among the intrinsic polypeptides specificfor —S plants. These plasma membrane polypeptides mightbe involved in sulphate uptake. Key words: Sulphate, sulphur-starvation, plasma membrane, polypeptides, root, transport     

Origin and development of river zooplankton: example of the Marne

Zooplankton composition and growth in the river Marne (France)were studied on a space scale of 300 km in July 1991. There werethree distinct areas along the river: the immediate reservoir outlet(pK 652), the natural river called middle Marne (from pK 652to pK 799 downstream) and the channeled river (from pK 799 topK 975 downstream). A typical lake community, characterized both byan abundance of microcrustaceans and a high zooplanktonconcentration was found immediately downstream of the reservoir Marne(Der-Chantecocq Lake). Here, large microcrustaceans (copepods,daphnids), and large rotifers (Keratella cochlearisrobusta and Polyarthra dolichoptera-vulgaris) rapidlydisappeared, and small rotifer species (<120 m) dominated theplankton. Their populations (specially Keratella c. cochlearis)proliferated in the middle Marne as far as 100 km downstream(up to 288 ind. l-1) but were considerably reduced (20 to35 ind. l-1) where the river is channeled, algal resourcesdecline and turbidity increases.The dominance of small organisms such as rotifers, in riverplankton is assumed to be the result of fish predation on largezooplankton as well as of a short generation time which allowstheir in situ reproduction, in spite of a short residencetime of the water.