Striking phenotypic and functional differences in lamina propria lymphocytes from the large and small intestine of mice

Although intraepithelial T lymphocytes of the large intestine (LI) are known to differ from those of the small intestine (SI) in phenotype and function, differences in LI and SI lamina propria (LP) lymphocyte populations have not been clearly established. In this work we found striking phenotypic differences between SI and LI LP lymphocyte populations from Balb/c mice analyzed by flow cytometry. In the LI most lymphocytes were B cells and the predominant T cells were TCR-alpha beta+, CD8+. In contrast, in the SI most T lymphocytes were CD4+ expressing TCR-alpha beta+, although a higher proportion expressed TCR-gamma delta+ than in the LI. In T cells the expression of adhesion molecules and cytokines was also different between SI and LI. The proportion of LP T cells expressing alpha4beta7 and L-selectin was higher in the LI than in the SI; whereas a greater proportion of cells expressing alpha(E)beta7 were detected in the SI than in LI. Higher proportions of T cells expressing L-selectin and alpha4beta1 were detected in the intraepithelial compartment of the LI than that of the SI, whereas the number of T cells expressing alpha(E)beta7 was much higher in the SI than in the LI. The proportion of T cells spontaneously producing IL-2, IFN gamma, and IL-4 at the intraepithelial and lamina propria, in the small and large intestine, was different indicating that distinctive functional features exist in the lymphocyte populations residing at the different intestinal compartments.     

Nucleotidic variations of two captive groups of tepezcuintle, Agouti paca (Rodentia: Agoutidae), from two sites in Yucatan, Mexico

The objective of this work was to estimate the nucleotidic variation between two groups of tepezcuintles (Agouti paca) from the states of Campeche and Quintana Roo, Mexico and within members of each group. Blood samples were collected from eleven A. paca kept in captivity. DNA from leukocytic cells was used for Ramdom Amplification of DNA Polimorphism (RAPD). The primers three 5'-d(GTAGACCCGT)- 3' and six 5'-d(CCCGTCAGCA)- 3' were selected from de Amersham kit (Ready.To.Go. RAPD Analysis Beads, Amersham Pharmacia Biotech), because they produced an adequate number of bands. The electrophoretic pattern of bands obtained was analyzed using software for phylogenetic analysis based on the UPGMA method, to estimate the units of nucleotidic variation. The phylogenetic tree obtained with primer three reveals a dicotomic grouping between the animals from both states in the Yucatan Peninsula showing a divergent value of 1.983 nucleotides per hundred. Animals from Quintana Roo show a grouping with primer six; an additional grouping was observed with animals from Campeche. Nucleotidic variation between both groups was 2.118 nucleotides per hundred. The nucleotidic variation for the two primers within the groups from both states, showed fluctuating values from 0.46 to 1.68 nucleotides per hundred, which indicates that nucleotidic variation between the two groups of animals is around two nucleotides per hundred and, within the groups, less than 1.7 nucleotides per hundred.     

Increased expression and phosphorylation of the two S100A9 isoforms in mononuclear cells from patients with systemic lupus erythematosus: a proteomic signature for circulating low-density granulocytes

Proteins differentially expressed in peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients versus Normal controls were identified by 2-DE and MALDI-MS. Thus, S100A9 expression was significantly increased in SLE PBMCs relative to Normal PBMCs at both mRNA and protein levels. Increased S100A9 levels in SLE PBMCs correlated positively with the abnormal presence of low-density granulocytes (LDGs) detected by flow-cytometry in the mononuclear cell fractions. Another set of proteins that were differentially expressed in SLE PBMCs formed S100A9-independent clusters, suggesting that these differences in protein expression are in fact reflecting changes in the abundance of specific cell types. In SLE PBMCs spots of the two S100A9 isoforms, S100A9-l and S100A9-s, and their phosphorylated counterparts were identified and confirmed to be phosphorylated at Thr¹¹³ by MS/MS analyses. In addition, the phorbol ester PMA alone or in combination with ionomycin induced a stronger increase in threonine phosphorylation of S100A9 in SLE than in Normal PBMCs, while the same stimuli caused the opposite effect on phosphorylation and activation of Erk1/2, suggesting the existence of an abnormal S100A9 signaling in SLE PBMCs. Therefore, the expansion and activation of LDGs in SLE seems to underlie this prominent S100A9 signature.     

Generation of a novel murine model of Aβ deposition based on the expression of human wild-type amyloid precursor protein gene

Mouse models of Alzheimer disease (AD) have been generated based on Amyloid-β Precursor Protein (AβPP) and the Presenilin (PSEN) gene mutations associated with familial AD (FAD). Such models have provided valuable insights into AD pathogenesis and represent an important research tool for the discovery of potential treatments. To model amyloid deposition in AD, we generated a new mouse line based on the presence of two copies of the genomic region encoding human wild-type AβPP as well as a mutation (L166P) in the murine Psen1. By ~6 months of age, these mice have begun to develop cerebral Aβ pathology with a significant increase in the levels of AβPP C-terminal fragments and Aβ42, as well as increase Aβ42/Aβ40 ratio. Since in the brain and other tissues of these mice, wild-type human AβPP mRNA and protein levels are comparable to those of endogenous AβPP, this model may allow studies about the role of AβPP isoforms in the pathogenesis of AD. This animal model may be suitable to test drugs aimed at inhibiting expression or altering splicing and processing of AβPP, without artifacts associated with the presence of mutations in AβPP or overexpression due to the use of exogenous promoters. These features of the new model are of critical importance in assessing the success of therapeutic interventions.     

The metal-binding features of the recombinant mussel <Emphasis Type="Italic">Mytilus edulis</Emphasis> MT-10-IV metallothionein

In contrast with the paradigmatic mammalian metallothioneins (MTs), mollusc MT systems consist at least of a high-cadmium induced form, possibly involved in detoxification, and another isoform either constitutive or regulated by essential metals and probably associated with housekeeping metabolism. With the aim of providing a deeper characterization of the coordination features of a molluscan MT peptide of the latter kind, we have analyzed here the metal-binding abilities of the recombinant MeMT-10-IV isoform of Mytilus edulis (MeMT). Also, comparison with other MTs of this type has been undertaken. A synthetic complementary DNA was constructed, cloned and expressed into two Escherichia coli systems. Upon zinc coordination, MeMT folds in vivo into highly chiral and stable Zn(7) complexes, with an exceptional reluctance to fully substitute cadmium(II) and/or copper(I) for zinc(II). In vivo cadmium binding leads to homometallic Cd(7) complexes that structurally differ from any of the in vitro prepared Cd(7) complexes. Homometallic Cu-MeMT can only be obtained in vitro from Zn(7)-MeMT after a great molar excess of copper(I) has been added. In vivo, two different heterometallic Zn,Cu-MeMT complexes are recovered, which nicely correspond to two distinct stages of the in vitro zinc/copper replacement. These MeMT metal-binding features are consistent with a physiological role related to basal/housekeeping metal, mainly zinc, metabolism, and confirm the correspondence between the MeMT gene response pattern and the functional properties of the encoded protein.     

Estimating plant responses to climate by direct gradient analysis and geographic distribution analysis

We characterised the climatic behaviour of 53 woody species in terms of the climatic factors that play the main role in controlling species distribution in the study area. Floristic and climatic data were obtained from 150 stands in sites under climatic control (i.e. eu-climatopes). The sampling strategy used allowed a reliable match between floristic and climatic observations. Different methods of frequency analysis and goodness-of-fit tests were used to identify associations between species occurrence and climatic characteristics. The species' responses were summarised by statistics describing ecological preferences and amplitudes, and species were grouped accordingly. A Gaussian response model was fitted to the abundance data along the main climatic gradients for selected species and response surfaces were derived by spatial analysis for a set of indicator species. Frequency analysis methods detected 42 indicator taxa for the Baudiere's Qe drought index, and lower numbers, 34 and 22, respectively, for the mean minimum coldest-month temperature and the daily temperature range in the coldest month. Goodness-of-fit tests revealed a lower number of ecological profiles with statistically significant deviations from equidistribution. We discuss the relative performance of the different methods and suggest that the combined use of statistical tests and frequency analyses may improve estimation of the environmental requirements of species. We also recommend using the species' responses to key environmental factors as reliable criteria in the definition of plant functional types. This revised version was published online in August 2006 with corrections to the Cover Date.     

Amazonian mangroves – A multidisciplinary case study in Pará State,North Brazil: Introduction

It is always a pleasant but not alwaysso easy task to write an introduction -ashort one- on the subject of Amazônia.The region is so rich in nature andculture, history and myths, that thedifficulty is to keep these aspectscompletely separated from each other, as inany interdisciplinary research.Fortunately, the layout of this edition hasleft a bit of extra space in thisintroduction for, besides the numbers andfacts, some etymology, discovery voyagesand old maps ¨.