An initial top-down proteomic analysis of the standard cuprizone mouse model of multiple sclerosis

An initial proteomic analysis of the cuprizone mouse model to characterise the breadth of toxicity by assessing cortex, skeletal muscle, spleen and peripheral blood mononuclear cells. Cuprizone treated vs. control mice for an initial characterisation. Select tissues from each group were pooled, analysed in triplicate using two-dimensional gel electrophoresis (2DE) and deep imaging and altered protein species identified using liquid chromatography tandem mass spectrometry (LC/MS/MS). Forty-three proteins were found to be uniquely detectable or undetectable in the cuprizone treatment group across the tissues analysed. Protein species identified in the cortex may potentially be linked to axonal damage in this model, and those in the spleen and peripheral blood mononuclear cells to the minimal peripheral immune cell infiltration into the central nervous system during cuprizone mediated demyelination. Primary oligodendrocytosis has been observed in type III lesions in multiple sclerosis. However, the underlying mechanisms are poorly understood. Cuprizone treatment results in oligodendrocyte apoptosis and secondary demyelination. This initial analysis identified proteins likely related to axonal damage; these may link primary oligodendrocytosis and secondary axonal damage. Furthermore, this appears to be the first study of the cuprizone model to also identify alterations in the proteomes of skeletal muscle, spleen and peripheral blood mononuclear cells. Notably, protein disulphide isomerase was not detected in the cuprizone cohort; its absence has been linked to reduced major histocompatibility class I assembly and reduced antigen presentation. Overall, the results suggest that, like experimental autoimmune encephalomyelitis, results from the standard cuprizone model should be carefully considered relative to clinical multiple sclerosis.     

Right lower lobe oedema as a rare sign of left heart failure

Two cases of right lower lobe oedema are presented as a supposedly very rare sign of left heart failure after myocardial infarction in the presence of acute mitral regurgitation. Causes of unilateral oedema and its clinical relevance in the diagnostic and therapeutic perspectives are discussed, and a review of the literature on this topic is presented.     

Conserved water-mediated H-bonding dynamics of catalytic Asn 175 in plant thiol protease

The role of invariant water molecules in the activity of plant cysteine protease is ubiquitous in nature. On analysing the 11 different Protein DataBank (PDB) structures of plant thiol proteases, the two invariant water molecules W1 and W2 (W220 and W222 in the template 1PPN structure) were observed to form H-bonds with the O_b atom of Asn 175. Extensive energy minimization and molecular dynamics simulation studies up to 2 ns on all the PDB and solvated structures clearly revealed the involvement of the H-bonding association of the two water molecules in fixing the orientation of the asparagine residue of the catalytic triad. From this study, it is suggested that H-bonding of the water molecule at the W1 invariant site better stabilizes the Asn residue at the active site of the catalytic triad.     

Using Haplotype Trees for Phylogeographic and Species Inference in Fish Populations

Genetic variation is now routinely screened at the DNA sequence level in many studies. If the DNA region being screened has not experienced excessive amounts of recombination, it is often possible to reconstruct the evolutionary history of the genetic variation in the form of a haplotype tree. This tree estimates the evolutionary pathway that interconnects all the different haplotypes (sequence variants) observed in the sample. This haplotype tree can be used to define a series of nested branches (clades) that reflects the relative temporal history of the haplotypes and groups of haplotypes. Geographical information can then be overlaid upon this temporal series to test for significant associations between geography and temporal position in the haplotype tree. This allows a reconstruction of how the genetic variation arose and spread in both space and time. Such reconstructions can yield many insights into the joint roles of recurrent events such as gene flow and of historical events such as fragmentation or range expansion. These points are illustrated with studies on the chub, Leuciscus cephalus. There is also a need to extend such nested phylogeographic analyses to a phylo/reticulate geographic analysis that incorporates both assortment and recombination between and within DNA regions. A preliminary phylo/reticulate geographic analysis is presented of the transferrin locus in the brown trout, Salmo trutta, species complex that reveals the importance of hybridization in the recent evolutionary history of this group. This example shows the inadequacy of a strictly phylogenetic approach and illustrates the need to incorporate reticulate evolution. The results of nested clade phylogeographic analysis and the new phylo/reticulate geographic analysis are then used for inferring species status of the marbled trout. The results indicate that an old hybridization event may have played a role in the origin of the marbled trout. Currently the marbled trout is primarily endangered by hybridization with introduced brown trout. These results show both the positive and negative impacts of hybridization upon biodiversity. Such phylo/reticulate geographic studies will challenge both our concepts of species and our conservation management strategies.     

O-acetyl-salicylic acid promotes colony formation from protoplasts of an elite maize inbred

The salicylic acid derivative acetylsalicylic acid (ASA) was found to promote colony formation from protoplasts isolated from embryogenic suspension cultures of an elite maize inbred line. The drug was most effective at concentrations of 30–100 mg/l, and increases of more than 20-fold in the number of colonies recovered from protoplasts were obtained. The rate of growth of protoplast-derived cell colonies was not affected.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - DMSO dimethylsulfoxide - KM Kao and Michayluk medium (1975) - MES 2[N-morpholino]ethanesulfonic acid - ASA acetylsalicylic acid     

Interleukin inhibition by a parasite proteinase inhibitor, taeniaestatin

A proteinase inhibitor, taeniaestatin, isolated from the larval stage of the cestode Taenia taeniaeformis inhibits endogenous IL 2 generation in murine lymphocytes and IL 1 induced proliferation of murine thymocytes in a dose-dependent manner. However, taeniaestatin does not inhibit exogenous IL 2-induced proliferation of an IL 2-dependent cell line at any dose tested. These data indicate that the lack of IL 2 generation may be due in part to inhibition of a crucial cell-associated proteinase subsequent to cellular activation, or the lack of an effective IL 2 signal for differentiation. Our results are novel findings concerning molecular pathways for parasite inhibition of host immune responses, and suggest that selected proteinase inhibitors may be useful in clinical situations in which IL 1 or IL 2 are elevated.