A proteomic approach to the identification of new tPA receptors in pancreatic cancer cells

We have developed a strategy to identify putative tissue-type plasminogen activator (tPA)receptors present in pancreatic cancer cells by affinity capture with tPA-Sepharose followed by 2-DE and MALDI-MS PMF. Proteins pulled down from either total lysates or raft membrane fractions were characterized and compared with those from a total lysate of an endothelial cell line (HUVEC) to identify pancreas-restricted tPA receptors. A total of 31 proteins were found by this approach, including annexin A2, already described as a tPA receptor in pancreas and endothelial cells, other proteins acting as tPA receptors (i.e., enolase, cytokeratins 8 and 18) in other tissues, and additional proteins not previously identified as candidate tPA receptors. Confirmation of the results was performed for some of these proteins using immunoblotting. These studies are the basis for further functional analyses on the role of these proteins in the biological effects of tPA.     

Land use and environmental factors affecting red-legged partridge (Alectoris rufa) hunting yields in southern Spain

The red-legged partridge is a small game species widely hunted in southern Spain. Its commercial use has important socioeconomic effects in rural areas where other agrarian uses are of marginal importance. The aims of the present work were to identify areas in Andalusia (southern Spain) where game yields for the red-legged partridge reach high values and to establish the environmental and land use factors that determine them. We analysed 32,134 annual hunting reports (HRs) produced by 6,049 game estates during the hunting seasons 1993/1994 to 2001/2002 to estimate the average hunting yields of red-legged partridge in each Andalusian municipality (n=771). We modelled the favourability for obtaining good hunting yields using stepwise logistic regression on a set of climatic, topographical, land use and vegetation variables that were available as digital coverages or tabular data applied to municipalities. Good hunting yields occur mainly in plain areas located in the Guadalquivir valley, at the bottom of Betic Range and in the Betic depressions. Favourable areas are related to highly mechanised, low-elevation areas mainly dedicated to intensive dry crops. The most favourable areas predicted by our model are mainly located in the Guadalquivir valley.     

A comparison of univariate and multivariate gene selection techniques for classification of cancer datasets

Background  

Gene selection is an important step when building predictors of disease state based on gene expression data. Gene selection generally improves performance and identifies a relevant subset of genes. Many univariate and multivariate gene selection approaches have been proposed. Frequently the claim is made that genes are co-regulated (due to pathway dependencies) and that multivariate approaches are therefore per definition more desirable than univariate selection approaches. Based on the published performances of all these approaches a fair comparison of the available results can not be made. This mainly stems from two factors. First, the results are often biased, since the validation set is in one way or another involved in training the predictor, resulting in optimistically biased performance estimates. Second, the published results are often based on a small number of relatively simple datasets. Consequently no generally applicable conclusions can be drawn.     

Assessing the potential range expansion of the exotic monk parakeet in Spain

In this study we determine favourable areas for the monk parakeet, Myiopsitta monachus , in peninsular Spain to account for its current distribution and predict its future course according to its potential range. We applied a favourability function based on generalized linear models using the presence/absence of breeding colonies of the species and the values of a set of variables on the 5167 UTM 10 × 10 km squares comprising the study area. We calculated the factor of distribution change in presences predicted by the model, and grouped the variables into explanatory factors performing a variation partitioning to assess the explanatory power of each factor. Our model included six predictors to explain the presence and absence of the species. These predictors were grouped into three factors: human activity, climate, and topography. Purely human influences accounted for 63.8% of the variation of the final model, while topographical variables explained 15.2% and climate only 5.7%. We obtained a high distribution change factor in which the presences of the species were predicted to increase between two- and sevenfold. Taking into account highly favourable squares, we conclude that the species is still absent in more than 72% of potential settlement areas, and thus we expect a continuous increase in the distribution of the species. Human activity is the main force moulding the distribution of the species, and lies behind its fast expansion, which is not only active, but is also passive via releases and escapes. We identified the areas of likely future expansion of the exotic monk parakeet in Spain. The pest status of the species in its native range, together to its distribution trend, should be taken into account by wildlife agencies to consider options for management.     

Cryptococcal lymphadenitis in a dog.

We describe a case of canine crytococcosis, the clinical symptoms were: feverish syndrome, vomiting and diarrhoeas and bilateral lymphadenitis in superficial lymph nodes. Microbiology and histopathology study of popliteal lymph node biopsy demonstrated the presence of round yeasts of some 3 microm of diameter, which we identified as Cryptococcus neoformans. Thirty months after suspending the medication the dog returned to the surgery; the dog was very thin and it had nervous symptoms; the owners decided it upon euthanasia. After carrying out the necropsy we take samples for their microbiology and histopathology study, both techniques detect to C. neoformans in marrow, CNS tissue and lymph nodes.     

Standard errors of heritabilities for forage breeding nurseries

Estimators for standard errors of heritability for two typical and complex forage breeding models are illustrated with estimates of variance and covariance components from least-squares and restricted maximum likelihood methods. The first experiment (Model 1) is a factorial design (two grazing management factors) with two layers of split plots (populations and grazing periods) and with plants within populations nested beneath the first split. The second model is for a sites pooling of randomized complete block experiments, with plants nested inside plots and with a split in time. One definition of heritability was applied to the plant habit character for both models in a red clover germplasm. Heritability estimates were statistically the same for the two methods for both models.     

Role of toxin activation on binding and pore formation activity of the Bacillus thuringiensis Cry3 toxins in membranes of Leptinotarsa decemlineata (Say)

Binding and pore formation constitute key steps in the mode of action of Bacillus thuringiensis delta-endotoxins. In this work, we present a comparative analysis of toxin-binding capacities of proteolytically processed Cry3A, Cry3B and Cry3C toxins to brush border membranes (BBMV) of the Colorado potato beetle Leptinotarsa decemlineata (CPB), a major potato coleopteran-insect pest. Competition experiments showed that the three Cry3 proteolytically activated toxins share a common binding site. Also heterologous competition experiments showed that Cry3Aa and Cry3Ca toxins have an extra binding site that is not shared with Cry3Ba toxin. The pore formation activity of the three different Cry3 toxins is analysed. High pore-formation activities were observed in Cry3 toxins obtained by proteolytical activation with CPB BBMV in contrast to toxins activated with either trypsin or chymotrypsin proteases. The pore-formation activity correlated with the formation of soluble oligomeric structures. Our data support that, similarly to the Cry1A toxins, the Cry3 oligomer is formed after receptor binding and before membrane insertion, forming a pre-pore structure that is insertion-competent.     

Oxygen deficiency affects carbohydrate reserves in overwintering forage crops

Anaerobic conditions developing under an ice cover affect winter survival and spring regrowth of economically important perennial crops. The objective was to compare, during a prolonged period of low (<2%) O2 at low temperature, the concentration of carbohydrates of four plant species contrasting in their resistance to oxygen deficiency. Four perennial forage species, lucerne (Medicago sativa L.), red clover (Trifolium pratense L.), timothy (Phleum pratense L.), and cocksfoot (Dactylis glomerata L.) were subjected to a progressively developing oxygen deficiency stress by enclosing potted plants in gas-tight bags in late autumn for overwintering in an unheated greenhouse. Timothy was previously reported to be more resistant to oxygen deficiency than the three other species. Non-structural carbohydrates increased and remained at a higher concentration in timothy than in the other three species under low O2 concentration. Concentrations of sucrose, fructose, glucose, and fructans increased in response to oxygen deficiency in timothy, whereas the concentration of soluble sugars decreased under the same conditions in lucerne, red clover, and cocksfoot. The gene expression of glyceraldehyde-3-phosphate dehydrogenase increased in response to low oxygen concentration in oxygen deficiency-sensitive lucerne while it remained unchanged in the oxygen deficiency-resistant timothy. It is concluded that timothy maintains higher carbohydrate reserves under oxygen deficiency, a specific feature that could favour its winter survival and spring regrowth.     

Pathogens: raft hijackers

Throughout evolution, organisms have developed immune-surveillance networks to protect themselves from potential pathogens. At the cellular level, the signalling events that regulate these defensive responses take place in membrane rafts--dynamic microdomains that are enriched in cholesterol and glycosphingolipids--that facilitate many protein-protein and lipid-protein interactions at the cell surface. Pathogens have evolved many strategies to ensure their own survival and to evade the host immune system, in some cases by hijacking rafts. However, understanding the means by which pathogens exploit rafts might lead to new therapeutic strategies to prevent or alleviate certain infectious diseases, such as those caused by HIV-1 or Ebola virus.