Repetitive DNA sequences located in the central region of the human mdr1 (multidrug resistance) gene may account for a gene fusion event during its evolution

The mdr1 gene, first member of the human multidrug-resistance gene family, is a major gene involved in cellular resistance to several drugs used in anticancer chemotherapy. Its product, the drug-excreting P-glycoprotein, shows a bipartite structure formed by two similar adjacent halves. According to one hypothesis, the fusion of two related ancestral genes during evolution could have resulted in this structure. The DNA sequence analysis of the introns located in the region connecting the two halves of the human mdr1 gene revealed a highly conserved poly(CA) · poly (TG) sequence in intron 15 and repeated sequences of the Alu family in introns 14 and 17. These repeated sequences most likely represent molecular fossils of ancient DNA elements which were involved in such a recombination event. Correspondence to: M. Pauly     

gamma-Carboxyglutamic acids 36 and 40 do not contribute to human factor IX function.

The gamma-carboxyglutamic acid (Gla) domains of the vitamin K-dependent blood coagulation proteins contain 10 highly conserved Gla residues within the first 33 residues, but factor IX is unique in possessing 2 additional Gla residues at positions 36 and 40. To determine their importance, factor IX species lacking these Gla residues were isolated from heterologously expressed human factor IX. Using ion-exchange chromatography, peptide mapping, mass spectrometry, and N-terminal sequencing, we have purified and identified two partially carboxylated recombinant factor IX species; factor IX/gamma 40E is uncarboxylated at residue 40 and factor IX/gamma 36,40E is uncarboxylated at both residues 36 and 40. These species were compared with the fully gamma-carboxylated recombinant factor IX, unfractionated recombinant factor IX, and plasma-derived factor IX. As monitored by anti-factor IX:Ca (II)-specific antibodies and by the quenching of intrinsic fluorescence, all these factor IX species underwent the Ca(II)-induced conformational transition required for phospholipid membrane binding and bound equivalently to phospholipid vesicles composed of phosphatidylserine, phosphatidylcholine, and phosphatidylethanolamine. Endothelial cell binding was also similar in all species, with half-maximal inhibition of the binding of 125I-labeled plasma-derived factor IX at concentrations of 2-6 nM. Functionally, factor IX/gamma 36,40E and factor IX/gamma 40E were similar to fully gamma-carboxylated recombinant factor IX and plasma-derived factor IX in their coagulant activity and in their ability to participate in the activation of factor X in the tenase complex both with synthetic phospholipid vesicles and activated platelets. However, Gla 36 and Gla 40 represent part of the epitope targeted by anti-factor IX:Mg(II)-specific antibodies because these antibodies bound factor IX preferentially to factor IX/gamma 36,40E and factor IX/gamma 40E. These results demonstrate that the gamma-carboxylation of glutamic acid residues 36 and 40 in human factor IX is not required for any function of factor IX examined.     

The impact of the Scheldt input on the trace metal distribution in the Belgian coastal area (results of 1981–1983 and 1995–1996)

Offshore fluxes of Cu, Zn, Cd, Pb and Hg were calculated based onresidual flow patterns and salinity gradients along the Belgian coast. Theresidual flow lines along the Belgian coast are more or less parallel to thecoast except in the area where the north-easterly flowing watermass comingfrom the Channel encounters the south-westerly-oriented Scheldt outflow,forming a residual hydrodynamical front. From the steady-state salinitypattern, diffusion coefficients perpendicular to the residual flow werededuced; they ranged from 21 to 108 m² s^-1. Offshore fluxes of dissolved and particulate trace metals based on diffusiveand mixing processes are calculated. The steady state profiles of dissolvedmetals show a dilution effect in the coastal waters, reaching an almostconstant concentration in the marine watermass in the 1981–1983dataset. The ratios of the Scheldt input of trace metals to the totaldissolved offshore flux vary from 38 to 55% (1981–1983),depending on the kind of metal, and from 55 to 91% (1995–1996).The ratio of the Scheldt input to the dissolved metal flow parallel to thecoast, is in both periods (1981–1983 and 1995–1996), smallerthan 1%. The steady-state concentration profiles of particular metalsversus salinity are fairly constant in the coastal-estuarine and marinewatermasses, but decrease very abruptly from the first to the secondwatermass. Assuming a conservative behaviour of the particular metals,offshore fluxes and the resulting concentration increases agree fairly wellwith the observed values. The ratios of the Scheldt input to the particulatetrace metal offshore flux vary between 30 to 46% (1981–1983)and 13 to 37% (1995–1996). The contribution of the Scheldtestuary to the flows parallel to the coast ranges from 1.6 to 2.9%(1981–1983) and from 0.6 to 1.6% (1995). This revised version was published online in July 2006 with corrections to the Cover Date.     

Endothelin Stimulates Phospholipase D in Striatal Astrocytes

Abstract: In primary cultures of mouse striatal astrocytes prelabeled with [³H]myristic acid, endothelin (ET)-1 induced a time-dependent formation of [³H]phosphatidic acid and [³H]diacylglycerol. In the presence of ethanol, a production of [³H]phosphatidylethanol was observed, indicating the activation of a phospholipase D (PLD). ET-1 and ET-3 were equipotent in stimulating PLD activity (EC₅₀ = 2–5 n M ). Pretreatment of the cells with pertussis toxin partially abolished the effect of ET-1, indicating the involvement of a G_i/G_o protein. Inhibition of protein kinase C by Ro 31-8220 or down-regulation of the kinase by a long-time treatment with phorbol 12-myristate 13-acetate (PMA) totally abolished the ET-1-induced stimulation of PLD. In contrast, a cyclic AMP-dependent process is not involved in the activation of PLD, because the ET-1-evoked formation of [³H]phosphatidylethanol was not affected when cells were coincubated with either isoproterenol, 8-bromo-cyclic AMP, or forskolin. Acute treatment with PMA also stimulated PLD through a protein kinase C-dependent process. However, the ET-1 and PMA responses were additive. Furthermore, the ET-1-evoked response, contrary to that of PMA, totally depended on the presence of extracellular calcium. These results suggest that at least two distinct mechanisms are involved in the control of PLD activity in striatal astrocytes. Finally, ET-1, ET-3, and PMA also stimulated PLD in astrocytes from the mesencephalon, the cerebral cortex, and the hippocampus.     

Modeling the population dynamics of annual plants with seed bank and density dependent effects

A model is proposed for the population dynamics of an annual plant (Sesbania vesicaria) with a seed bank (i.e. in which a proportion of seeds remain dormant for at least one year). A simple linear matrix model is deduced from the life cycle graph. The dominant eigenvalue of the projection matrix is estimated from demographic parameters derived from field studies. The estimated values for population growth rate () indicates that the study population should be experiencing a rapid exponential increase, but this was not the case in our population.The addition of density dependent effects on seedling survivorship and adult fecundity, effects for which field studies provide evidence, considerably improves our model. Depending on the demographic parameters, the model leads to stable equilibrium, oscillations, or chaos. Study of the behaviour of this model in the parameter space shows that the existence of a seed bank allows higher among-year variation of adult fecundity, without leaving the region of demographic stability. Field data obtained over 3 years confirm this prediction.     

Polynucleotide sequence relatedness among motileAeromonas species

Relatedness among 55 motileAeromonas strains was assessed by determining the extent of reassociation in heterologous DNA preparations. S1 nuclease and diethylaminoethyl-cellulose filters were used to separate reassociated nucleotide sequences from nonreassociated sequences and to determine the thermal stability of related nucleotide sequences. The motile aeromonads would presently consist of three species:A. hydrophila (type strain ATCC 7966),A. caviae (type strain ATCC 15468), andA. sobria (type strain CIP 7433). These three species are clearly separated on the basis of both biochemical characteristics and similarities in DNA. Each of these three species contains more than one hybridization group: three groups inA. hydrophila; two groups inA. caviae; and at least two groups inA. sobria. DNA hybridization groups are biochemically similar within each species. When additional data is available, these separate DNA hybridization groups may merit designation. Any decision to delineate species in terms of DNA hybridization groups must await a phenotypic basis for their separation.     

Kinetics of induction of error-prone repair of bacteriophage lambda by temperature shift in an Escherichia coli dnaB mutant.

Summary Preincubation at 42^o, before infection at permissive temperature by phage , of an Escherichia coli dnaB mutant, provokes a significant increase in survival and mutagenesis of ultraviolet irradiated phage as well as mutagenesis of untreated phage. Similarly to UV irradiation and many chemical mutagens, the inhibition of DNA synthesis by temperature shift of this dnaB mutant induces SOS repair. This work shows that replication blockage in bacterial DNA is not only mutagenic for bacterial DNA itself (Witkin, 1975) but also for normally replicating DNA, probably due to induction of diffusible products.     

Variations in oxygen consumption of Demidoff's bushbaby (Galago demidovii, lorisidae, primates) in captivity

Summary In captivity,Galago demidovii shows annual variations in oxygen consumption which are independent of daylength cycles. This rhythm is characterized by two periods during which metabolic and sexual activities are increased: a short period in November/December; and a longer period from March to June inclusive. These two periods alternate with periods of decreased metabolic rate, of which the most pronounced extends from July to September. Another point of interest is that the basal metabolism of captive Galagos is 17.5% above the value calculated from its body weight.The physiological cycle observed in captivity is synchronized with climatic variations in Makokou (Gaboon), from where the animals originated: higher metabolic and sexual activities are correlated with rainy seasons. This study suggests that an endogenous rhythm may exist inGalago demidovii.     

Biochemical studies carried out on different groups ofCandida parapsilosis andCandida rhagii strains by comparing some cellular and mitochondrial activities

A comparative biochemical study was performed on some strains ofCandida rhagii and on strains belonging to different subgroups ofCandida parapsilosis. Measurements of alcohol dehydrogenase activity, resistance to drugs and occurrence of an alternative pathway enabled us to confirm the classification between several subgroups within theC. parapsilosis species.