Drosophila melanogaster acetylcholinesterase: Identification and expression of two mutations responsible for cold- and heat-sensitive phenotypes

Ace ^IJ29 and Ac ^IJ40 are cold- and heat-sensitive variants of the gene coding for acetylcholinesterase in Drosophila melanogaster. In the homozygous condition, these mutations are lethal when animals are raised at restrictive temperatures, i.e., below 23° C for Ace ^IJ29 or above 25° C for Ace ^IJ40. The coding regions of the gene in these mutants were sequenced and mutations changing Ser³⁷⁴ to Phe in Ace ^IJ29 and Pro⁷⁵ to Leu in Ace ^IJ40 were found. Acetylcholinesterases bearing these mutations were expressed in Xenopus oocytes and we found that these mutations decrease the secretion rate of the protein most probably by affecting its folding. This phenomenon is exacerbated at restrictive temperatures decreasing the amount of secreted acetylcholinesterase below the lethality threshold. In parallel, the substitution of the conserved Asp²⁴⁸ by an Asn residue completely inhibits the activity of the enzyme and its secretion, preventing the correct folding of the protein in a non-conditional manner.     

Separation of the enantiomers of coumarinic anticoagulant drugs by capillary electrophoresis using maltodextrins as chiral modifiers

The separation and quantitation of coumarinic anticoagulant drug enantiomers were achieved by direct chiral capillary electrophoresis using complex maltooligosaccharide mixtures as stereoselective electrolyte modifiers. Chiral separations were characterized by a high selectivity and efficiency, enabling enantiomeric excess determinations. In addition, preliminary results indicate the applicability of the method for the determination of individual enantiomers in biological samples. So the method can be used to perform stereoselective pharmacokinetic studies. © 1994 Wiley-Liss, Inc.     

Environmental NGOs,dispossession, and the state: The ideology and praxis of African nature and development

Zimbabwe provides a significant context to examine the interplay of the new development rhetoric, the actions of powerful conservation organizations, donor policies, a relatively strong and stable government, and the empowerment of local communities. This interplay exists in diverse rural areas where the Communal Areas Management Program for Indigenous Resources (CAMPFIRE) is in various stages of experimentation and implementation. CAMPFIRE has been described as a philosophy of sustainable rural development that enables rural communities to manage, and benefit directly form indigenous wildlife. It is the best known of African efforts to permit African communities to re- assert their management of selected natural resources. The program has the official support of the Zimbabwean government. Nonetheless, there are many potential areas of serious conflict. Three case studies are utilized to explore how the challenges of repossession of critical environmental resources by marginalized communities in the changing context of state and NGO relationships where international tourism is a revenue generator for both the private sector and government.     

Enhancement of radiation-induced cell killing and DNA double-strand breaks in a human tumor cell line using nanomolar concentrations of aclacinomycin A.

Several agents that induce differentiation have previously been shown to induce the terminal differentiation of leukemic cells and enhance the radiosensitivity of certain solid tumor cell lines in vitro using millimolar concentrations. We now report that aclacinomycin A (ACM), a potent inducer of leukemic cell differentiation in vitro, can significantly enhance the radiosensitivity of a human colon tumor cell line (Clone A) at a concentration of 10 nM. Based on colony-forming efficiency, the maximum increase in radiosensitivity was found using 15 nM ACM for 3 days with a dose enhancement factor of 1.4 at a surviving fraction of 0.10. This treatment increased cell doubling time, but had no effect on cell-cycle phase distribution. To gain insight into the mechanisms responsible for this radiosensitization, gamma-ray-induced DNA single- and double-strand breaks were examined. Aclacinomycin A had no effect on the induction of DNA single-strand breaks but significantly enhanced the formation of gamma-ray-induced DNA double-strand breaks. The rate or extent of repair of the induced double-strand breaks was not influenced by ACM treatment. These data suggest that ACM, at achievable plasma concentrations, can enhance the radiosensitivity of a human tumor cell line by increasing the initial level of radiation-induced DNA double-strand breaks.     

3-Deoxy-D-arabino heptulosonate 7-phosphate synthase from Zea mays: General properties and regulation by tryptophan

In extracts from Zea mays shoots, the presence of thiol compoundsin the extraction buffer was necessary to get an active 3 deoxy-D-arabinoheptulosonic acid 7-phosphate (DAHP) synthase. Its pH optimumfor activity was about 7.5. Of the different cations tested,only Mn⁺⁺ was an activator. Enzyme stability was optimal inTris-HCl buffer, pH 7.5, that contained a reducing agent, Mn⁺⁺and a polyol. Contrary to other reports, phosphoenolpyruvate(PEP) did not stabilize the preparation significantly. The synthaseexhibited high affinities for both erythrose-4-phosphate (Km:0.24 mM) and PEP (Km: 0.31 mM). Its specific activity was highestin young shoots. Corn DAHP synthase was inhibited in vitro by tryptophan. Moreover,the enzyme was retarded on a tryptophan agarose affinity column,but it was removed with the bulk of protein from the same supportwhen eluted with buffer containing tryptophan. Inhibition whichwas easily lost during storage at 4°C was pH dependent andincreased during development. Maximal inhibition, about 60%with 1 mM tryptophan, was observed in extracts from 8 day-oldshoots. Phenylalanine and tyrosine were not inhibitory, andno synergistic effects were observed when the aromatic aminoacids were tested in combination. Isoenzymes could not be demonstrated. (Received April 23, 1980; )     

L'âge du complexe récifal des côtes de meuse entre verdun et commercy et la stratigraphie de l'oxfordien dans l'est du bassin de Paris

The question of the age of «Hauts de Meuse coral rag is studied again using new faunas of brachiopods and ammonites collected above, below and inside the coral rag. Ammonites fauna of the well known «Calcaires de Creuë is revised. Results obtained agree closely.The «Hauts de Meuse coral rag grew up during Middle Oxfordian time; and extend part of the Plicatilis zone (Antecedens Subzone) and the whole Transversarium Zone up to the basis of the Bifurcatus Zone, for the first time identified in Lorraine. So it span a short depositional time, about 1 MY, with a high deposition rate, the thickness of the coral rag being more than 100 meters, reaching 150 meters in Saint-Mihiel area. Towards SE, on the other side of the Marne Gap, the sudden change into basinal facies go with a reducing of thickness and deposition rate.Upper Oxfordian deposits show a relative uniformity of facies and thickness. At the top of Middle Oxfordian limestones, the age of the discontinuity and the Sequanian renewal of deposition is basal Bifurcatus Zone. Erosional renewal and terrigeneous deposits are a wide spread event by the beginning of Upper Oxfordian.In the paleontological part, species upon which are based new ages and interpretations are only figured, except the species new or with new paleontological data, which are described. Type specimen of Perisphinctes mosensisBayle, from the «Calcaires de Creuë, is described and photographically figured for the first time.