Molecular dynamics study of the dissociation of an antigen-antibody complex in solution

Preliminary results are reported of a molecular dynamics calculation of free energy variations during the dissociation of an antigen-antibody complex, hen egg-white lysozyme — Fab D1.3, using atomic coordinates determined by the group of R. J. Poljak, and explicit handling of solvent molecules. After equilibration of the complex in solution at 300 K, a dissociation path was generated by a directed dynamics protocol. Then the thermodynamic perturbation method was used for computing the derivative of the free energy of the system with respect to dissociation coordinate, both for the undissociated complex and in two points along the path. 200-ps molecular dynamics simulations were carried out at each of these points. The results obtained are discussed, with special emphasis on the role of interstitial water in the appearance of a hydrophobic activation free energy.Université Paul Sabatier, and URA 505 of C.N.R.S.     

In Rhizobium meliloti, the operon associated with the nod box n5 comprises nodL, noeA and noeB, three host-range genes specifically required for the nodulation of particular Medicago species

In Rhizobium meliloti , the genes required for nodulation of legume hosts are under the control of DNA regulatory sequences called nod boxes. In this paper, we have characterized three host-specific nodulation genes, which form a flavonoid-inducible operon down-stream of the nod box n5. The first gene of this operon is identical to the nodL gene identified by Baev and Kondorosi (1992) in R. meliloti strain AK631. The product of the second gene, NoeA, presents some homology with a methyl transferase. nodL mutants synthesize Nod factors lacking the O -acetate substituent. In contrast, in strains carrying a mutation in either noeA or noeB , no modification in Nod-factor structure or production could be detected. On particular hosts, such as Medicago littoralis , mutants of the n5 operon showed a very weak nodule-forming ability, associated with a drastic decrease in the number of infection threads, while nodulation of Medicago truncatula or Melilotus alba was not affected. Thus, nodL , noeA and noeB are host-specific nodulation genes. By using a gain-of-function approach, we showed that the presence of nodL , and hence of O -acetylated Nod factors, is a major prerequisite for confering the ability to nodulate alfalfa upon the heterologous bacterium Rhizobium tropici .     

Variations in maturation strains and root shape in root systems of Maritime pine (Pinus pinaster Ait.)

 In order to determine if different types of wood were being laid down in the root system of Maritime pine (Pinus pinaster Ait), in response to wind loading, longitudinal residual maturation strains (LRMS), indicating the existence of mechanical stress in developing wood cells, were measured in the trunk and lateral roots. Two age groups of trees (5- and 13-year- old) were compared. LRMS were greater in the trunk and roots of 13-year-old trees than in 5-year-old trees. This phenomenon may be due to increased competition between older trees. LRMS in leeward roots of both age-groups were positive i.e. the wood cells had developed under compression, as also occurs in reaction wood of gymnosperms. As leeward roots are placed under compression during tree sway, an abnormal type of wood may form in the roots in order to counteract the increased stress. In other roots, the strains were negative i.e. the cells had developed under tension, as occurs in normal wood. In the roots of younger trees, LRMS were also positive nearer the stem, thus indicating that wood formation may also be influenced by bending stresses experienced in this zone. In addition to LRMS measurements, radial growth in roots was examined in order to determine the influence of mechanical loading on secondary growth. In older trees, there was a significant increase of 34% in woody growth below the biological centre, compared to that above. This eccentricity is unlike that found in most other tree species, where secondary growth is usually greater on the upper side of the root. However, Maritime pine has a tap root, which will alter the pattern of stress within the root system. Under wind loading, a concentration of mechanical stress will develop at the bases of the stem, lateral roots and tap root. Received: 7 July 1997 / Accepted: 11 December 1997     

Rhizobial lipochitooligosaccharide nodulation factors activate expression of the legume early nodulin gene ENOD12 in rice

Lipochitooligosaccharide nodulation factors (Nod factors) produced by rhizobia are a major host range determinant. These factors play a pivotal role in the molecular signal exchange, infection and induction of symbiotic developmental responses in legumes leading to the formation of a nodule in which rhizobia carry out N₂ fixation. Determining whether rice ( Oryza sativa ) can respond to Nod factors could lead to strategies that would make rice amenable to develop a nitrogen-fixing endosymbiotic association with rhizobia. We introduced into rice the promoter of the infection-related gene MtENOD12 (from Medicago truncatula ) fused to the β-glucuronidase (GUS) reporter gene to serve as a molecular marker to aid in the detection of Nod factor signal perception by rice cells. Treatment of the transgenic rice roots with Nod factors (10^–6–10^–9 m ) under nitrogen-limiting conditions induced MtENOD12 -GUS expression in cortical parenchyma, endodermis and pericycle. In contrast, chitooligosaccharide backbone alone failed to elicit such a response in the root tissues. These findings demonstrate that rice roots perceive Nod factors and that these lipochitooligosaccharides, but not simple chitin oligomers, act as signal molecules in activating MtENOD12 in cortical parenchyma as in legumes. Exogenous application of N -naphthaleneacetic acid mimicked the Nod factor-elicited tissue-specific expression of MtENOD12 in roots while cytokinins inhibited it, thus evidencing that Nod factors, auxin and cytokinins probably act on similar signaling elements responsible for the regulation of MtENOD12 activation in rice. Taken together, these results suggest that at least a portion of the signal transduction machinery important for legume nodulation is likely to exist in rice.      

Obtaining Vitis Vinifera cell cultures producing higher amounts of malvidin-3-O-β-glucoside

To obtain a cell line of Vitis vinifera producing mainly malvidin-3-O--glucoside, three methods were studied: the initiation of new cell cultures from different varieties, the modification of phytohormones in the culture media of a high anthocyanin-producing cell line, and the selection by small-aggregate cloning of new cell lines from this high producing strain. The latter technique gave a cell line accumulating a malvidin-3-O--glucoside proportion of 63%.     

Oxidative cross-linking of chemically and enzymatically modified sugar-beet pectin

Hot acid-soluble pectins from sugar-beet pulp which do not gel in the presence of persulfate were submitted to hydrolysis with acid under different conditions of concentration, temperature and time, or with different combinations of enzymes. All the modified pectins have been chemically characterised and tested for their gelling capacity with persulfate ions. They varied primarily in the structure of the side-chains and only those obtained from cold acid hydrolysis and from degradation by arabino-furanosidase were able to gel. Accessibility of the feruloyl groups carried by the arabinose side-chains appeared essential for the gelling of beet pectins with persulfate.     

Interaction between aflatoxin B1 and oxytetracycline in isolated rat hepatocytes

Isolated rat hepatocytes were used as an in vitro model to investigate A possible interaction between oxytetracycline (OXT) and aflatoxin B₁ (AFB₁). LDH leakage, RNA and protein synthesis and glycogen accumulation were measured in the presence of both drugs, either separately or in combination. The evolution of LDH leakage during the incubation was identical in untreated and treated cells. AFB₁ inhibited RNA and protein synthesis at a concentration of 10^–7 M and 10^–6 M, respectively, and higher, whereas OXT did not influence RNA synthesis but inhibited protein synthesis at the highest tested concentration, 10^–3 M. As far as glycogen is concerned, rats were injected with glucagon before sacrifice in order to obtain a constant synthesis rate in isolated hepatocytes. AFB₁ inhibited the accumulation of glycogen from 10^–6 M upward. This effect was never observed before 90 min of incubation. OXT had no effect on glycogen synthesis. In the presence of both drugs, no interaction was demonstrated as far as RNA and protein synthesis were concerned, but OXT opposed the inhibition induced by AFB₁ on glycogen accumulation. If the in vivo protection, provided by OXT against AFBI-induced toxicity, is due to a direct interference in the toxic mechanisms of the mycotoxin, these results show that OXT does not influence the AFB₁-inhibition of RNA and protein synthesis. The latter are early and sensitive parameters inhibited by AFB₁. On the contrary, taking into consideration the results on glycogen accumulation, it seems more interesting to investigate further this metabolism.Abbreviations AFB₁ Aflatoxin B₁ - OXT Oxytetracycline - DMEM Dulbecco's Modified Eagle's Medium - LDH Lactate Dehydrogenase - DMSO Dimethyl Sulfoxide - BSA Bovine Serum Albumin