Intercalation of anthracyclines into living cell DNA analyzed by flow cytometry.

Anthracyclines (ANT) are used in the treatment of leukemia and other cancers. These drugs have been shown to intercalate between the strands of DNA. In the present study, we show that the amount of ANT intercalated into DNA can be determined by measuring the fluorescence resonance energy transfer (FRET) between Hoechst 33342 (H33342) and ANT bound to DNA. The transfer efficiency was found to depend on the amount of disposable ANT but was independent of the amount of H33342 bound to DNA over a wide range of H33342 concentrations. The method was adapted for flow cytometric measurement of FRET in whole living cells and was used to evaluate the degree of intercalation of daunorubicin (DAU) and idarubicine (IDA) into DAU-sensitive and DAU-resistant leukemic cell lines. ANT intercalation into DNA was affected by factors which modify the intracytoplasmic concentration of ANT, and it was shown that the action of ANT and the resistance to ANT could not be attributed solely to the intercalative effect of the drugs. The method has advantages over previously described methods and represents a useful complementary tool in studies on the mode of action of ANT and the mechanisms of chemoresistance.     

Recognition of HLA-B27 by mouse cytotoxic T-cell clones: a transgenic mouse

As a basis for the characterization of mouse T cells involved in the recognition of xenogeneic HLA molecules, a panel of HLA-B27-reactive cytotoxic T-cell clones was generated upon stimulation by cells from HLA-B27-transgenic mice. The HLA-B27-induced T-cell response was found to comprise two categories of clones: some recognizing HLA-B27 independent of H-2 molecules expressed by the target cells (unrestricted clones), others recognizing HLA-B27 in an H-2 restricted manner. The unrestricted clones exhibited diverse specificities, as judged from their various cross-reactivities with other xenogeneic (HLA) or allogeneic (H-2) molecules. In addition, although most of the unrestricted clones were able to react with both mouse and human HLA-B27-transgenic mice. The HLA-B27 induced T-cell which reacted only with HLA-B27-positive mouse, and not human cells. These findings illustrate that both H-2-restricted and unrestricted T cells with diverse species contribute to HLA-B27-xenorecognition.     

Differentiation of keratinocytesin vitro: a new culture vessel mimicking thein vivo situation

A culture vessel consisting of two independent chambers separated only by the growth substrate is described. Cells may be cultured on both sides of the growth substrate. Culture medium and gas exposure can independently be controlled in both compartments. Human hair follicles have been used as source of keratinocytes and the bovine eye lens capsule has been explored as growth substrate. The presence of 5% CO₂ in air in the lower compartment appears to have a significant effect on the morphology of the cultures. When the cultures are being exposed to air with 5% CO₂, the culture medium being applied in the lower compartment, formation of corneocytes characteristic for adult stratum corneum is induced, as evidenced by light and electron microscopy. To the knowledge of the authors, this stage of differentiationin vitro has not been obtained with previously described systems. Differentiation of the lower cell layers has been characterised with specific antibodies. The possible use of the system for applied and pure scientific research is discussed.     

Theoretical conformational analysis of phospholipids II. Role of hydration in the gel to liquid crystal transition of phospholipids

To obtain a satisfactory agreement between computed transition temperatures and those determined experimentally, we introduce explicitly water molecules which hydrate the polar headgroup of dipalmitoylphosphatidylethanolamine molecules. The calculated free energy curves as a function of the intermolecular interchain distance and the degree of hydration of the polar groups permit the determination of the transition of the phospholipid system from the gel to the liquid crystalline phase. The detailed structure of the hydration shell is defined using the supermolecular approach.     

Les supports morphologiques de l'intégration dans la colonie de Veretillum cynomorium Pall. (Cnidaria,Penntularia)

The morphological supports of integration, including the nervous system, are to be found.After a morphological observation, ecto and endoderms and their intercellular junctions are presented.The regionally different mesogleal synaptic nerve nets are surrounded by ecto and endodermal nervous structures.The presence of behavioural activity centers is also discussed.

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Liste des abréviations des figures AS autozoïde sectionné - AU autozoïde - CAL canal longitudinal - CAV cavité gastrique - CC corps compact - CGL cellule globuleuse myo-épitheliale MESlame mésentériale MF microfilament - CGR cellule granuleuse myo-épithéliale - CLC cloison commune à deux autozoïdes - CLR cloison radiale - CLT cloison transversale - CM corps myélinique - CME cellule mesenchymateuse - CNE cellule nerveuse ectodermique - CNEN cellule nerveuse endodermique - CNM cellule nerveuse mésogléenne - COL colonne - CSP cellule spumeuse - CSPH cellule sphéruleuse - CU cellule urticante - D dépot de matériel intercellulaire (zonula adhaerens) - DS desmosome septé - ECT ectoderme - EE empilement d'ergastoplasme rappelant un corps de Nissl - FF fibres fasciculées h structure - G glycogène - GA granule aréolé - IC inclusion crênelée - M mitochondrie - MB membrane basale - ME mésoglée - MES lame mésentériale - MF microfilament - MT figure interprétée on terme de microtubule - MV microvilli - N noyau - E neurite - NO nodosité - P pédoncule - PC paroi des canaux - PE perforation - R rachis - SF siphonozoïde fendu - SIPH siphonozoïde - SM sipment musculaire d'une cellule myoépithéliale - T travé ous-siphonozoïdale - TME travée mésogléenne - VC vésicule claire - VME vésicule mésenchymateuse - ZI zône intermédiaire - I synapse polarisée - 2 synapse réciproque - 3 figure ressemblant à un desmosome septé - 4 contact par apposition Cc travail correspond à la première partie d'une thèse de Doctorat d'Etat, consacrée aux «Structures et aux mécanismes de l'intégration dans la colonie de Veretillum cynomoriurra» — Il a été effectué au sein de l'Equipe de Recherche Associée au C.N.R.S., n°183 (Directeur Al. Pavans de Ceccatty) avec le concours du Laboratoire Arago à Banyuls sur Mer (France) et la collaboration technique de Madame J. Villeneuve.