In vivo isolation of a pPJ3a-λ cosmid mediated by the Tn2307 transposon

From an E. coli cell harboring plasmid pPJ3b (= pPJ3a::Tn2301) and infected with phage λ, we have isolated two defective phages having inserted pPJ3a DNA and Tn2301 in their genomes. One of them has been extensively characterized: it behaves like a cosmid, i.e., upon injection into the cell, its DNA circularizes and replicates as a plasmid (pPJ10); it can be packaged again in λ heads, provided the presence of a phage helper. Furthermore, heteroduplex analysis has shown that in pPJ10, the transposon Tn2301 is inverted compared to its direction in pPJ3b. We give evidence suggesting that this type of inversion is in part mediated by Tn2301.     

Régulation de la biosynthèse de l'aspartate-ammonium lyase (aspartase) de Pseudomonas fluorescens

Variations in aspartasic activity in various media are due to aspartate-ammonium lyase induction and to regulation of the biosynthesis of this enzyme. Evidence for neosynthesis of the enzyme is provided by labelling and separation of the protein. The inducer appears to be aspartic acid. The biosynthesis is subject to pronounced catabolic repression. The physiological function of aspartate-ammonium lyase is discussed.     

Methyl-4 formyl-7 cyclopenta(c)pyranne isole apres hydrolyse acide de Viburnum tinus

Viburtinal (4-methyl-7-formylcyclopenta(c)pyrane), yet unknown, was obtained by acid hydrolysis of the esters extracted from Viburnum tinus. Its structure was deduced by spectroscopic methods.     

Evolution de la musculature desNéréidiens (Annélides polychètes) au cours de l'Epitoquie

Résumé Au cours de l'épitoquie des Nereidiens, les fibres musculaires longitudinales ne sont pas forméesde novo, à partir de cellules indifferenciées ou myoblastes, mais proviennent des fibres anciennes atoques. Celles-ci subissent une véritable dédifférenciation plus ou moins synchrone d'une redifférenciation. Les deux processus ne sont pas successifs mais simultanés, et une dédifférenciation complète est absente.Les premières cellules en évolution appartiennent à la couche musculaire externe; ensuite, les fibres des assises plus profondes se transforment à leur tour.Les transformations consistent en: 1) La dédifférenciation du bord interne ou coelomique de la fibre. Les structures contractiles disparaissent dans cette zone et de nombreuses particules de glycogène se différencient sans relation avec le reticulum endoplasmique ou les ribosomes. Aucun lysosome ou signe précurseur ne peuvent être observés avant la disparition des filaments contractiles et des éléments Z. 2) Le bord coelomique s'hypertrophie. Dans la région axiale de la fibre, de nombreuses mitochondries et particules et de glycogène remplacent le matériel contractile. Corrélativement, l'épaisseur des bandes A et I diminue. 3) La fibre hétéronéreidienne ou épitoque est constituée et présente deux parties: un cortex myoplasmique et une médulla sarcoplasmique, remplie de mitochondries et de glycogène. Le noyau renfermant un nucléole volumineux est situé dans une hernie sarcoplasmique latérale.

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Evolution of muscles inNereidae (Annelida polychaeta) during Epitoky. III. Dedifferentiation of the longitudinal fibres

Summary During epitoky inNereidae, the longitudinal muscle fibres are not formedde novo from undifferentiated cells or myoblasts, but arise from the old atokous fibres. These undergo a true dedifferentiation more or less synchronously with a redifferentiation. The two processes are not successive but simultaneous and there is no complete dedifferentiation.The first cells that develop are in the outside muscle layer; then the fibres of the inside layers are transformed in their turn.The transformations consist of: 1) Dedifferentiation of the edge of the inner or coelomic fibre. The contractile structures disappear in this part and numerous glycogen particles differentiate, unrelated to endoplasmic reticulum or ribosomes. No lysosomes or precursory markings are observed before the disappearance of contractile filaments and Z rods. 2) The coelomic edge becomes enlarged. In the axial region of the fibre, numerous mitochondria and and glycogen particles take the place of the contractile material. Consequently, the thickness of A and I bands decreases. 3) The heteronereid or epitokous fibre is formed and shows two parts: a myoplasmic cortex and a sarcoplasmic medulla, filled with mitochondria and glycogen. The nucleus with a voluminous nucleolus settles inside a lateral sarcoplasmic swelling.

     

Regioelectronic factors in metabolic hydroxylation of aliphatic carbon atoms

EHT calculations have been performed on model molecules acting as substrates for mammalian mono-oxygenases. C_α---H bonds are consistently found to have larger overlap populations compared with C_β---H and C_γ---H bonds. It is known on the other hand that metabolic hydroxylation of aliphatic carbon atoms shows a marked regioselectivity for α-carbons. The quantum-mechanical results sustain the view that C---H bonds of relatively high electronic density are preferred target sites for the cytochrome P-450-mediated oxygenation, and that the oxygen atom being activated is transformed into an electrophilic species capable of C---H bond insertion.     

Influence of dicarboxylic phosphatidylcholines on phosphatidylcholine liposomes as revealed by gel chromatography and electron microscopy

The effect of dicarboxylic phosphatidylcholines (glutarylphosphatidylcholine) on the structural changes of phosphatidylcholine liposomes is examined by using multilamellar liposomes prepared with egg phosphatidylcholine or dipalmitoylphosphatidylcholine and by varying the surface charge by addition of dicetyl phosphate. Investigations are performed by gel chromatography and electron microscopy. Glutarylphosphatidylcholine is in micellar form (rod-like micelles or globular micelles). The structures obtained depend on the fatty acid saturation of liposomes and on the charge of liposome (addition or not of dicetyl phosphate). With egg phosphatidylcholine/glutarylphosphatidylcholine dispersions, an aspect more similar to myelinic figures than liposomes is observed, while in the presence of dicetyl phosphate, liposomes similar to control egg phosphatidylcholine liposomes are obtained. Gel chromatography on Sepharose 4B and turbidity measurements prove that dicetyl phosphate increases the stability of egg phosphatidylcholine/glutarylphosphatidylcholine mixtures. On the other hand, in dipalmitoylphosphatidylcholine/glutarylphosphatidylcholine dispersions, incorporation of dicetyl phosphate destabilizes bilayer structure and the formation of mixed micelles occurs. Viscosity measurement shows, in the presence of dicetyl phosphate, an increased fluidity for dipalmitoylphosphatidylcholine/glutarylphosphatidylcholine dispersions, in agreement with the micellar organization. These data confirm that the disorganization of liposomal membranes by dicarboxylic phosphatidylcholine depends on the fatty acid composition of phosphatidylcholine and on the presence of dicetyl phosphate.     

Characterization of messenger RNA for aldolase B in adult and fetal human liver

High molecular weight cellular RNA was isolated from adult and fetal human liver tissue by a procedure of ethanol precipitation in concentrated guanidine-HCl solutions. About 5 mg of RNA were obtained from one gram of liver. RNA was fractionated by sucrose gradient ultracentrifugation. Aldolase B neosynthesized in a reticulocyte lysate cell-free system under the direction of total or fractionated RNA was purified by immunoaffinity microchromatography. Messenger RNA specifying synthesis of aldolase B exhibited a sedimentation coefficient of 16 S both in adult and fetal liver. This enzyme represented 1.3 % of the total neosynthesized proteins in adult liver, 0.1 % in the liver of a 6-month-old fetus and less than 0.01 % in the liver of a 4.5 month-old fetus.     

Mitochondrial DNA evolution in lagomorphs: Origin of systematic heteroplasmy and organization of diversity in European rabbits

Summary A characterization was conducted on mitochondrial DNA (mtDNA) molecules extracted separately from 107 European rabbits (Oryctolagus cuniculus) both wild and domestic, 13 European hares (Lepus capensis), and 1 eastern cottontail (Sylvilagus floridanus). Experimentally this study took into account restriction site polymorphism, overall length variation of the noncoding region, and numbers of repeated sequences. Nucleotide divergences indicate that the mtDNAs from the three species derived from a common ancestor some 6–8 million years (Myr) ago. Every animal appeared heteroplasmic for a set of molecules with various lengths of the noncoding region and variable numbers of repeated sequences that contribute to them. This systematic heteroplasmy, most probably generated by a rate of localized mtDNA rearrangements high enough to counterbalance the cellular segregation of rearranged molecules, is a shared derived character of leporids.The geographic distribution of mtDNA polymorphism among wild rabbit populations over the western European basin shows that two molecular lineages are represented, one in southern Spain, the second over northern Spain, France, and Tunisia. These two lineages derived from a common ancestor some 2 Myr ago. Their present geographical distribution may be correlated to the separation of rabbits into two stocks at the time of Mindel glaciation.Finally the distribution of mtDNA diversity exhibits a mosaic pattern both at inter- and intrapopulation levels.