On the need for combining complementary analyses to assess the effect of a candidate gene and the evolution of its polymorphism: the example of the Major Histocompatibility Complex in chicken

The aim of this paper is to combine different but complementary approaches to check the neutrality of a given locus in a selected population. Analysis was undertaken through the polymorphism's evolution compared with that predicted under the effect of drift and through the analysis of the variance components of the measured traits, considering the effect of the locus as either a fixed or a random effect. This study deals with the case of the MHC locus, using both data from experimental lines of chicken selected for three different criteria of immune response, and frequencies of the genotyped haplotypes over time. Both the evolution of the polymorphism and the variance components approach have led to the conclusion that the MHC locus has an effect on the trait affecting antibody production against the Newcastle disease virus. Results have also highlighted the interest in using various methods in the case of low allelic frequencies. However, none of the common hypotheses, overdominance or frequency-dependent selection, was sufficient to explain the observed variation of the MHC polymorphism, which was displayed by the temporal variation of the allelic frequencies.     

The induction of reciprocal translocations in mouse germ cells by chemicals and ionizing radiations. II. Combined effects of mitomycin C or thio-tepa with two doses of gamma-rays

The combined effects of mitomycin C (MMC) and thio-tepa (TT) with gamma-ray doses of 5 and 9 Gy on mouse stem cells were studied using the spermatocyte test. Both chemicals induced very low yields of translocations after single treatments. In combined treatments with a dose of 5 Gy, a subadditive effect of MMC and an additive effect of TT were found. Combined with a dose of 9 Gy the compounds potentiated the effect of radiations. Up to now, most of the chemicals tested have shown additive effects when combined with doses of the ascending part of the dose-response curve and potentiating effects when combined with doses of its descending part. This has been considered additional confirmation of the concept that depletion of any kind of spermatogonia is sufficient to modify the genetic response of stem cells. However, the subadditive and additive responses found could be considered evidence that common biological mechanisms can modulate the response to combined treatments of chemicals and ionizing radiations.     

Modifications by ischemia of carbohydrate and lipid metabolism of the dog heart in situ from circulating and endogenous substrates

The uptake of circulating substrates, lactate, glucose and free fatty acids (FFA) has been investigated concurrently with the tissular contents of these principles and the glycogen and triglyceride stores in the dog heart in situ submitted to incomplete obstruction of left coronary bed. Transmural samples necessary for the repeated determination of tissular substrates were taken from left ventricular wall by means of a total cardiopulmonary by-pass system, then divided to allow the analysis separately in subendocardial and subepicardial layer. A 40 to 70% reduction in coronary blood flow gave rise to decrease or suppression of uptake of all the substrates or even to conversion of uptake into output. The modifications of uptake are chiefly related to the deficiency of breakdown by oxidation, though lessened in the case of FFA by incorporation into triglycerides and enhanced in the case of glucose by glycogenolysis. Glycogenolysis and consequent anaerobic glycolysis appear to be the main process available against the energy cellular defect linked with oxygen lack which affects notably more subendocardial than subepicardial layer.     

Daunorubicin-induced apoptosis: triggering of ceramide generation through sphingomyelin hydrolysis.

The nature of the signaling pathway(s) which initiate drug-triggered apoptosis remains largely unknown and is of fundamental importance in understanding cell death induced by chemotherapeutic agents. Here we show that in the leukemic cell lines U937 and HL-60, daunorubicin, at concentrations which trigger apoptosis, stimulated two distinct cycles of sphingomyelin hydrolysis (approximately 20% decrease at 1 microM) within 4-10 min and 60-75 min with concomitant ceramide generation. We demonstrate that the increase in ceramide levels, which precedes apoptosis, is mediated by a neutral sphingomyelinase and not by ceramide synthase. Indeed, potent ceramide synthase inhibitors such as fumonisin B1 did not affect daunorubicin-triggered sphingomyelin hydrolysis, ceramide generation or apoptosis. In conclusion, we provide evidence that daunorubicin-triggered apoptosis is mediated by a signaling pathway which is initiated by an early sphingomyelin-derived ceramide production.     

The Structural Basis for the Integrity of Adenovirus Ad3 Dodecahedron

During the viral life cycle adenoviruses produce excess capsid proteins. Human adenovirus serotype 3 (Ad3) synthesizes predominantly an excess of free pentons, the complexes of pentameric penton base and trimeric fiber proteins, which are responsible for virus penetration. In infected cells Ad3 pentons spontaneously assemble into dodecahedral virus-like nano-particles containing twelve pentons. They also form in insect cells during expression in the baculovirus system. Similarly, in the absence of fiber protein dodecahedric particles built of 12 penton base pentamers can be produced. Both kinds of dodecahedra show remarkable efficiency of intracellular penetration and can be engineered to deliver several millions of foreign cargo molecules to a single target cell. For this reason, they are of great interest as a delivery vector. In order to successfully manipulate this potential vector for drug and/or gene delivery, an understanding of the molecular basis of vector assembly and integrity is critical. Crystallographic data in conjunction with site-directed mutagenesis and biochemical analysis provide a model for the molecular determinants of dodecamer particle assembly and the requirements for stability. The 3.8 Å crystal structure of Ad3 penton base dodecamer (Dd) shows that the dodecahedric structure is stabilized by strand-swapping between neighboring penton base molecules. Such N-terminal strand-swapping does not occur for Dd of Ad2, a serotype which does not form Dd under physiological conditions. This unique stabilization of the Ad3 dodecamer is controlled by residues 59–61 located at the site of strand switching, the residues involved in putative salt bridges between pentamers and by the disordered N-terminus (residues 1–47), as confirmed by site directed mutagenesis and biochemical analysis of mutant and wild type protein. We also provide evidence that the distal N-terminal residues are externally exposed and available for attaching cargo.     

Caractérisation de complexes enzymes-antienzymes dans les extraits de levures du genre candida après immunoélectrophorégramme en agarose

Résumé Quatorze enzymes ont été identifiés parmi les fractions de l'antigène somatique deCandida albicans révélées sur immunoélectrophorégramme.Aucune activité enzymatique n'a cependant été jusqu'ici caractérisée au niveau des deux fractions qui, dans nos conditions expérimentales, s'étaient révélées antérieurement très spécifiques de cette espèce. Parmi les enzymes identifiés, la malicodéhydrogénase, fraction commune aux sept espèces étudiées (C. albicans, C. stellatoidea, C. tropicalis, C. zeylanoides, C. krusei, C. pseudotropicalis, C. macedoniensis) est la première à susciter la production de l'anticorps correspondant: elle s'est donc avérée la plus antigénique en fonction toutefois de son importance relative au sein de la mosaïque antigénique et compte tenu de la technique de détection des anticorps utilisée.     

Agar-supported cultivation of Halorubrum sp. SSR,and production of halocin C8 on the scale-up prototype Platotex

Halorubrum sp. SSR was isolated from a solar saltern in Algeria. The strain exhibited a high antibiotic activity against the indicator strain Natronorubrum aibiense G23, and the bioactive compound showed thermal, acid and alkali stability. SSR was grown on agar-supported cultivation (AgSF) to compare yields and applicability with traditional submerged cultivation. AgSF scale-up was implemented taking benefit from the solid-state cultivation prototype Platotex. This technology leads to high amounts of the target Halocin and facilitate the downstream steps. The antibiotic compound was purified according to a fast efficient procedure including ion exchange chromatography followed by a fractionation on C18 Sep-Pack cartridge. The compound was identified as Halocin C8 according to N-terminal amino acid sequencing and high-resolution mass spectrometry.