Solution structure of a DNA duplex containing the exocyclic lesion 3,N4-etheno-2'-deoxycytidine opposite 2'-deoxyguanosine

Vinyl chloride reacts with cellular DNA producing 3,N4-etheno-2'-deoxycytidine (epsilonC) along with other exocyclic adducts. The solution structure of an oligodeoxynucleotide duplex containing an epsilonC.dG base pair was determined by high-resolution NMR spectroscopy and molecular dynamics simulations. NMR data indicated that the duplex adopts a right-handed helical structure having all residues in anti orientation around the glycosylic torsion angle. The epsilonC adduct has a sugar pucker in the C3'-endo/C4'-exo region while the rest of the residues are in the C2'-endo/C3'-exo range. NOE interactions established Watson-Crick alignments for canonical base pairs of the duplex. The imino proton of the lesion-containing base pair resonated as a sharp signal that was resistant to water exchange, suggesting hydrogen bonding. Restrained molecular dynamics simulations generated three-dimensional models in excellent agreement with the spectroscopic data. The refined structures are slightly bent at the lesion site without major perturbations of the sugar-phosphate backbone. The adduct is displaced and shifted toward the major groove of the helix while its partner on the complementary strand remains stacked. The epsilonC(anti).dG(anti) base pair alignment is sheared and stabilized by the formation of hydrogen bonds. The biological implications of structures of epsilonC-containing DNA duplexes are discussed.     

Genetic mapping of a major susceptibility locus for juvenile myoclonic epilepsy on chromosome 15q

The epilepsies are a group of disorders characterised by recurrent seizures caused by episodes of abnormal neuronal hyperexcitability involving the brain. Up to 60 million people are affected worldwide and genetic factors may contribute to the aetiology in up to 40% of patients. The most common human genetic epilepsies display a complex pattern of inheritance. These are categorised as idiopathic in the absence of detectable structural or metabolic abnormalities. Juvenile myoclonic epilepsy (JME) is a distinctive and common variety of familial idiopathic generalised epilepsy (IGE) with a prevalence of 0.5-1.0 per 1000 and a ratio of sibling risk to population prevalence (lambda(s)) of 42. The molecular genetic basis of these familial idiopathic epilepsies is entirely unknown, but a mutation in the gene CHRNA4, encoding the alpha4 subunit of the neuronal nicotinic acetylcholine receptor (nAChR), was recently identified in a rare Mendelian variety of idiopathic epilepsy. Chromosomal regions harbouring genes for nAChR subunits were therefore tested for linkage to the JME trait in 34 pedigrees. Significant evidence for linkage with heterogeneity was found to polymorphic loci encompassing the region in which the gene encoding the alpha7 subunit of nAChR (CHRNA7) maps on chromosome 15q14 (HLOD = 4.4 at alpha = 0.65; Z(all) = 2.94, P = 0.0005). This major locus contributes to genetic susceptibility to JME in a majority of the families studied.     

Use of the polymerase chain reaction in the diagnosis of leprosy

A case is described in which a pericardial branch of a nongrafted left internal mammary artery communicated directly with the distal left anterior descending artery, following saphenous vein bypass grafting. This type of collateralization following coronary artery bypass surgery seems to be very rare, and perhaps could protect the myocardium from severe ischemia.     

Using Multimedia to Support Cooperative Editing

By Cooperative Editing we mean the coordinated manipulation of information by a group of authors. During the editing process the (co-)authors need to communicate their ideas, drafts and constraints (remotely or face-to-face) until a final version of the information is achieved. For the different phases of this process - discussion of ideas, editing, cross-checking - different media or media integration are adequate. Furthermore, analysing the transition from individual work to group work, within different human activities, two pitfalls are often detected if computer support is considered: a) technological communication difficulties, especially if the group is remotely located on heterogeneous hardware, associated with a fall in productivity and frequent social inadequacies of the group's computer support; b) the lack of integrated media processing tools available for group editing. In order to solve these problems, we suggest that Multimedia can be applied in two ways: to effectively support the necessary group communication links; and to enhance the expressiveness of the information edited. To test this statement we have been conceptualising and implementing a prototype system. Most of the techniques involved can be used in other tools that need multimedia capabilities or that support other specific types of group activities. The innovative aspects of the work are the use of multimedia techniques to support demanding applications, possibly on cross-platforms, and the integration of several concepts to support cooperation.     

Cooperative Hypermedia Editing with CoMEdiA

CoMEdiA is a groupware tool which enables co-authors to cooperatively produce hypermedia documents.CoMEdiA allows co-authors to communicate their ideas,drafts,guidelines,constraints and annotations to other co-authors.In uses a mix of communication patterns,media and document organization to enable co-authors to keep on exchanging information(remotely or face-to-face),improving passages and modifying notes until a final document is achieved.We did not concentrate on th depth but on the breath of the features.Our efforts were on integrating and corrdinating concepts from collaboration,multimedia and hyper organization rather than on making a specialized system in any of them.We began with the text medium and are now including bitmaps and raster images.Later sound and video will also be integrated.In this paper we describe and sustain the available capacites.     

Optical and electrical characterization of fluorine doped cadmium oxide thin films prepared by the sol-gel method

Highly transparent and conducting fluorine (F) doped cadmium oxide (CdO) thin films were deposited on glass slides by the sol-gel method. The films were doped by the addition of ammonium fluoride to the precursor solution whose optimum concentration was determined. The films were fired in an open atmosphere at 350 °C and after that, exposed to annealing treatments in different atmospheres (N₂, N₂/H₂ mixture and Ar) at the same temperature. The films were characterized by ultraviolet-visible spectroscopy, X‐ray diffraction and scanning electron microscopy. The resistivity was determined by the four probes method and current-voltage measurements in accordance with the standard Van der Pauw configuration. The CdO:F thin films obtained, showed high polycrystalline quality and high transmission in the visible region (≥ 90%), shifting towards the blue region of the absorption edge as the fluorine concentration in the precursor solution was increased from 0 to 30 at.%. The lowest resistivity values were reached for the samples with F content higher or equal to 5% and annealed in either N₂ or a 96/4 N₂/H₂ gas mixture. Our resistivity value reached in the CdO:F layers was 4.5 × 10^{− 4} Ω cm (20 Ω/square).     

Liver glutathione content and glutathione-dependent enzymes of two species of freshwater fish as bioindicators of chemical pollution

Glutathione content and glutathione-dependent enzymes were measured in the liver of two fish species, gudgeon (Gobio gobio) and roach (Rutilus arcasii), from the river Bernesga (Spain) caught downstream and upstream of the waste site of several chemical industries. Animals from contaminated sites display a reduced glutathione concentration and a tendency to the decrease of glutathione S-transferase activity. Glutathione peroxidase activity was significantly elevated only in the liver of Gobio gobio and glutathione reductase activity in that of Rutilus arcasii. Our data indicate that the glutathione system constitutes a sensitive biochemical indicator of chemical pollution. Relative changes of glutathione and glutathione-dependent enzymes in both fish species suggest a different susceptibility to toxins.     

Epitope specificity of CD44 for monoclonal antibody-dependent facilitation of marrow engraftment in a canine model

Primary graft rejection after marrow transplantation occurs more frequently in patients receiving HLA-haploidentical compared with HLA-identical sibling transplants. Both human and experimental animal data suggest that the cells responsible for this phenomenon are either host natural killer (NK) cells, T cells, or both. To investigate the mechanisms of graft rejection, we have developed a canine model of marrow transplantation, which uses DLA-nonidentical unrelated donors in the absence of postgrafting immunosuppression. In this model most animals rejected their marrow grafts after a preparative regimen of 9.2 Gy total body irradiation (TBI). However, engraftment of DLA-nonidentical marrow can be facilitated when the recipients are pretreated with monoclonal antibody (MoAb) S5, which recognizes CD44. In this report, we extended these observations by first cloning the canine CD44 and, next, mapping the epitope recognized by S5, which was located in a region conserved among human and canine CD44 and was distinct from the hyaluronan binding domain. However, in vitro binding of S5 caused a conformational change in CD44, which allowed increased hyaluronan binding. Then, we reexamined the in vivo model of marrow transplantation and compared results with MoAb S5 to those with two other anti-CD44 MoAbs, IM7 and S3. Only MoAb S5 significantly increased the engraftment rate of DLA-nonidentical unrelated marrow, whereas the two other anti-CD44 MoAbs were ineffective. The enhanced in vivo effect was not related to differences in the MoAbs' avidities, since both S5 and IM7 had equivalent binding to CD44, but most likely related to the specific epitope that S5 recognizes. Thus, this study shows that the effect of the anti-CD44 MoAb S5 in facilitating engraftment is epitope specific and if one is to use an anti-CD44 to facilitate engraftment of marrow in humans, one cannot assume that any anti-CD44 would work.