Genetic control of the oil content in Thymus vulgaris L: a case of polymorphism in a biosynthetic chain

Six distinet chemotypes of the mediterranean Thymus vulgaris L. are characterized by differences in the major monoterpenes produced. The nature of the monoterpenes is shown to be controlled by a series of loci. Each locus probably has a dominant and a recessive allele. Epistatic relationships between these loci are interpreted by assuming that terpenes are end products of branches from the same biosynthetic pathway. The most dominant types, which correspond to the monoterpenes produced at the beginning of the chain, show the most important potential genetic diversity. There is good evidence that regulatory genos are involved in this system.     

Expression of the Saccharomyces cerevisiae Kex2p endoprotease in inset cells. Evidence for a carboxy-terminal autoprocessing event.

The pheromone-processing Kex2p endoprotease of Saccharomyces cerevisiae has been difficult to characterize due to its low level of expression in yeast cells. To overcome this problem, we have overexpressed Kex2p using the baculovirus/insect cell expression system. Spodoptera frugiperda Sf9 insect cells infected with a recombinant baculovirus, containing the complete KEX2 gene which encodes the Kex2p protease (814 amino acids), accumulate an 120-kDa functional form of the enzyme. The inhibition profile of the insect-cell-derived endoprotease is similar to that of the yeast enzyme. The recombinant infected insect cells also secrete into the medium about half of the total Kex2p activity produced. Deleting the carboxyl-terminal tail and the transmembrane domain of Kex2p (Kex2 delta p, 666 amino acids) does not measurably interfere with the enzyme characteristics and results in the secretion of up to 90% of the total enzyme activity. The truncated form, Kex2 delta p, of the endoprotease accumulates in the cell supernatant to 6.7 x 10(5) U/l. The molecular mass of the secreted forms for both the wild-type Kex2p and Kex2 delta p is the same (70 kDa) and is 50-kDa lower than the intracellular form. This result implicates a processing event which gives rise to shorter extracellular forms of both the wild-type Kex2p and Kex2 delta p and which trims their carboxy termini upsteam of amino acid 666. This processing event requires the integrity of the Ser385 of the Kex2p active site.     

Climatic and anthropogenic effects on the sedimentation and geochemistry of Lakes Bourget,Annecy and Léman

During the last glaciation, the Rhone glacier extended as far as the region of Lyon and covered Lakes Bourget, Annecy and Léman. Glacial retreat successively freed Lakes Bourget and Annecy, it reached Geneva around 14 000 B.P. and the head of Lake Léman at about 12 000 B.P. Deposits situated between Nyon and the foot of the Jura provide a complete palynological zonation and serve as a type section for the lake core studies. A palynological horizon (about 1 800 A.D.) has been found in Lake Bourget and wood has been dated at 3 230 ± 65 B.P. (¹⁴C) in a core from Lake Léman. Three 6 m cores from each lake were sampled together with many 0.5 m cores from Léman. The short Lake Léman cores have been dated by ¹³⁷Cs and provide a precise indication of changes in the basin during recent decades. Holocene climatic variations are evident in the Léman cores and are reflected by concentrations of carbonate and organic carbon which increase at the end of the Alleröd and reach a maximum in the Atlantic Period (climatic optimum). Human settlement on the shores of Lake Annecy is shown by increases in heavy metals; increased sedimentation rates suggest settlement at the beginning of the Christian era. All the cores show increases in the organic matter and nutrients; from the turn of the century in the Léman, and later for the other lakes where eutrophication started only a few decades ago. NAI-P shows a very recent increase, around 1965 in the Leman. Heavy metals (Hg, Cd and Pb) show increases at about 30 years ago in Annecy, at about the turn of the century in Lake Léman and in the mid-19th century in Lake Bourget.

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Résumé Lors de la dernière glaciation, le glacier du Rhône a atteint la région de Lyon et recouvert les lacs du Bourget, d'Annecy et le Léman. Son retrait a successivement libéré les lacs du Bourget, d'Annecy, puis le Léman de leur carapace de glace. Aux environs de 14 000 ans B.P. son front se trouvait au voisinage de Genéve, puis vers 12 000 B.P. à l'embouchure du Rhône en amont du Léman.La chronostratigraphie des grandes carottes (6 m) prélevées dans les trois lacs a été établie par la palynologie, sur la base des zonations obtenues dans un profil complet provenant d'un marais situé entre Nyon et le pied du Jura. Elles ont pu être contrôlées par un marqueur palynologique dans le lac du Bourget (150 B.P.) et la datation au ¹⁴C d'un fragment de bois dans le Léman (3 230 ± 65 B.P.). Une série de carottes de grand diamètre provenant de ce dernier lac, datés par le ¹³⁷Cs, ont donné une image très précise du l'évolution du milieu durant les dernières décades. D'une facon générale, toutes les carottes montrent une augmentation des teneurs en matière organique, puis en nutrients, vers le fin du 19ème siècle pour le Léman et quelques dizaines d'années plus tard pour les lacs du Bourget et d'Annecy. Le début du l'etrophisation se situe beaucoup plus tard dans le Lémen avec l'accroissement des teneurs en phosphore, principalement de ses formes NAI-P et O-P, soit aux environs de 1965.

     

In vitro reconstitution of a trimeric complex of DivIB, DivIC and FtsL, and their transient co-localization at the division site in Streptococcus pneumoniae

DivIB, DivIC and FtsL are bacterial proteins essential for cell division, which show interdependencies for their stabilities and localization. We have reconstituted in vitro a trimeric complex consisting of the recombinant extracellular domains of the three proteins from Streptococcus pneumoniae. The extracellular domain of DivIB was found to associate with a heterodimer of those of DivIC and FtsL. The heterodimerization of DivIC and FtsL was artificially constrained by fusion with interacting coiled-coils. Immunofluorescence experiments showed that DivIC is always localized at mid-cell, in contrast to DivIB and FtsL, which are co-localized with DivIC only during septation. Taken together, our data suggest that assembly of the trimeric complex DivIB/DivIC/FtsL is regulated during the cell cycle through controlled formation of the DivIC/FtsL heterodimer.     

Characterizing the effect of endocrine disruptors on human health: The role of epidemiological cohorts

Research on endocrine disruptors (EDs) developed from numerous disciplines. In this concert of disciplines, epidemiology is central to inform on the relevance for humans of mechanisms and dose-response functions identified in animals, to characterize the health impact (number of attributable disease cases), the cost associated with ED exposure, and the efficiency of the measures taken to limit exposure. Here, we present epidemiological tools to draw valid inference regarding effects of potential EDs. Epidemiology is generally observational, requiring care to control confounding bias. Many potential EDs have a short biological half-life; approaches relying on repeated biospecimens sampling allow limiting exposure misclassification and the resulting bias. For non-persistent compounds, couple–child cohorts are a central study design. Cohorts can now rely on molecular biology approaches to characterize exposures and intermediate pathways, which corresponds to the advent of molecular epidemiology and allows stronger interactions between epidemiology, toxicology, and molecular epidemiology to characterize the health effects of EDs.     

Relationships of the Col plasmids E2, E3, E4, E5, E6, and E7: Restriction mapping and colicin gene fusions

Thirteen ColE plasmids representing the E2-E7 types have been compared by restriction mapping. Over 80% of their restriction sites were found to be similarly positioned, indicating that these plasmids share a common structure. Three variants are ColE2-CA42 and ColE7-K317, both of which contain 1.8-kb DNA segments in place of a 2.5-kb segment common to the other plasmids, and ColE6-CT14, which has an additional 5.0-kb DNA segment compared to the other plasmids. The colicin (col), immunity (imm), and colicin release (hic) genes of these plasmids have been localized to regions corresponding to those known for ColE3-CA38 and ColE2-P9, with the imm and hic genes adjacent to the 3' end of the col gene. Active colicin is produced from hybrid col genes containing 5' and 3' ends from different E-type plasmids. The 3'-termini of the fused col genes specify the colicin type.     

Gene therapy of erectile dysfunction in the rat with penile neuronal nitric oxide synthase

Gene transfer to the penile corpora cavernosa of constructs of the inducible and endothelial nitric oxide synthase (NOS) cDNAs ameliorates erectile dysfunction in aged rats. In this study, we investigated whether the neuronal NOS (nNOS) variant responsible for erection, penile nNOS (PnNOS), can exert a similar effect, and whether the combination of electroporation with a helper-dependent adenovirus (AdV) improves gene transfer. PnNOS and beta-galactosidase cDNAs were cloned in plasmid (pCMV-PnNOS; pCMV-beta-gal) and "gutless" AdV (AdV-CMV-PnNOS; AdV-CMV-beta-gal) vectors, and injected into the penis of adult (beta-gal) or aged (PnNOS) rats, with or without electroporation. Penile erection was measured at different times after PnNOS cDNA injection, by electrical field stimulation of the cavernosal nerve. The expression of beta-galactosidase or PnNOS was estimated in penile tissue by either histochemistry and luminometry or Western blot, and the effects of AdV-CMV-PnNOS on mRNA expression were examined by a DNA microarray. We found that electroporation increased pCMV-beta-gal uptake, and its expression was detectable at 56 days. In the aged rats treated with pCMV-PnNOS and electroporation, the maximal intracavernosal:mean arterial pressure ratios were elevated for 11 and 18 days when compared with those in controls. Electroporation intensified penile uptake of as few as 10(6) viral particles (vp) of AdV-CMV-beta-gal, and with 10(7) vp beta-galactosidase was still detectable at 60 days. Electroporated AdV-CMV-PnNOS (10(7) vp) was effective at 18 days in stimulating the erection of aged rats, without inducing the expression of cytotoxic genes. In conclusion, intracavernosal gene therapy with PnNOS cDNA corrected the aging-related erectile dysfunction for at least 18 days when given by electroporation in a helper-dependent AdV at low viral loads.