Ecology of cryptogamic epiphytes and their communities in deciduous forests in mediterranean Spain

A distribution of the epiphytic bryophyte and lichen vegetation of deciduous forests from the centre and north of the supramediterranean belt from the Spanish Iberian Peninsula was established using numerrical (PCA, CCA) and traditional floristic characteristics. Four groups have been established in relation to several syntaxa. Derived by numerical classification, possible successional trends between the groups with respect to changes in moisture, light and diameter of the trees are described. Nevertheless, patterns with succession in epiphytic communities are very complex to establish.     

Optimization of the conditions of the fermentation of beet molasses to lactic acid by Lactobacillus delbrueckii

Optimization studies were carried out for the production of L-lactic acid from the fermentation of beet molasses by Lactobacillus delbrueckii. A Central Composite Design was used to determine the optimum values of the process variables (temperature, pH, inoculum concentration, and initial sucrose concentration) for obtaining the maximum yield and the maximum volumetric productivity of lactic acid. Among the variables selected for study, it was found that all of them apart from the temperature significantly affected the responses (yield and volumetric productivity of lactic acid). The Central Composite Design also permitted formulating two second-order polynomial empirical models relating to the responses and the significant variables. From these models it was possible to determine the value of the variables giving the maximum yield of lactic acid production (87.8%) and the maximum volumetric rate of lactic acid biosynthesis (2.7 g/l · h). Finally, the dependence of the lactic acid yield and productivity on the model variables was investigated. All conclusions are restricted to the experimental range studied.     

Fine-scale geographical structure, intra-individual polymorphism and recombination in nuclear ribosomal internal transcribed spacers in Armeria (Plumbaginaceae)

BACKGROUND AND AIMS: Isolation and drift are the main causes for geographic structure of molecular variation. In contrast, the one found in a previous survey in Armeria (Plumbaginaceae) for nuclear ribosomal ITS multicopy regions was species-independent and has been hypothesized to be due to extensive gene-flow and biased concerted evolution. Since this was inferred from a genus-level phylogenetic analysis, the aim of this study was to check for the occurrence of such structure and the validity of the proposed model at a local scale, in a southern Spanish massif (Sierra Nevada), as well as to examine the evolutionary implications at the organism level. METHODS: In addition to 117 sequences of direct PCR products from genomic DNA, 50 sequences of PCR products from cloned DNA were obtained to analyse cases of intragenomic polymorphisms for the ITS regions. KEY RESULTS: Sequence data confirm the occurrence of a species-independent structure at a local scale and reveal insights through the analysis of contact areas between different ITS copies (ribotypes). A comparison between cloned and direct sequences (a) confirms that, within these contact areas, ITS copies co-occur both in different individuals and within single genomes; and (b) reveals recombination between different copies. CONCLUSIONS: This study supports the utility of direct sequences for detecting intra-individual polymorphism and for partially inferring the ITS copies involved, given previous knowledge of the variability. The main evolutionary implication at the organism level is that gene-flow and concerted evolution shape the geographic structure of ITS variation.     

Preparation of Single Cell Detritus from <Emphasis Type="Italic">Laminaria saccharina</Emphasis> as a Hatchery Diet for Bivalve Mollusks

A high-yield technique is described for the elaboration of single cell detritus (SCD) from Laminaria saccharina, based on the sequential action of C1H, enzymes (endoglucanases and cellulases) and 2 bacteria showing a high degree of cellobiotic, proteolytic, and alginolytic activity (CECT 5255 and CECT 5256). Over 85% of dried particles of L. saccharina were transformed into a suspension of free cell and bacterial and detrital particles after 24 hours of bacterial activity with this technique. These particles were less than 20 m in diameter, constituting a suitable diet for bivalve mollusks. After 72 hours 99% of the total particulate volume consisted of particles less than 20 m in diameter. Tests of hatchery diets for the seed of clam Ruditapes decussatus revealed increases of 54% and 68% for live weight and length, respectively, when SCD from L. saccharina was used as the sole dietary component compared with a live phytoplankton diet. However, SCD from L. saccharina is not a suitable food for the larvae of R. decussatus.     

Additive polymorphisms and reticulation in an ITS phylogeny of thrifts (Armeria,Plumbaginaceae)

A parsimony analysis of 133 sequences of the nuclear ribosomal DNA ITS1+5.8S+ITS2 region from 71 taxa in Armeria was carried out. The presence of additive polymorphic sites (APS; occurring in 14 accessions) fits the reticulate scenario proposed in previous work for explaining the ITS pattern of variation on a much smaller scale and is based mainly on the geographical structure of the data, irrespective of taxonomic boundaries. Despite the relatively low bootstrap values and large polytomies, part of which are likely due to disruptive effects of reticulation and concerted evolution in these multicopy sequences, the ITS analysis has phylogenetic and biogeographic implications. APS detected in this study are consistent with hypothesized hybridization events, although biased concerted evolution, previously documented in the genus, needs to be invoked for specific cases and may be responsible for a possible "sink" effect in terminals from a large clade. The causes for sequences of the same species appearing in different clades (here termed transclade) are discussed.     

Pentamidine is an antiparasitic and apoptotic drug that selectively modifies ubiquitin

We have determined the cytotoxic properties of pentamidine isethionate (2) towards the promastigotes of the protozoan parasite Leishmania infantum. The leishmanicidal activity of 2 was 60 times higher after 72 h of incubation than that of cisplatin (4). The pentamidine salt 2 induced a higher amount of programmed cell death (PCD) than cisplatin, which is associated with inhibition of DNA synthesis and cell-cycle arrest in the G2/M phase. Circular dichroism (CD) data indicate that binding of 2 to calf-thymus DNA (CT-DNA) induces conformational changes in the DNA double helix, consistent with a B-->A transition. Moreover, the interaction of 2 with ubiquitin led to a 6% increase in the beta-sheet content of the protein as observed by CD spectroscopy. Fluorescence-spectroscopy studies agreed with the CD data, showing that the pentamidine portion of 2 induces a significant decrease in the fluorescence of the Ub residues Phe4 and Phe45 located on the beta-cluster of the molecule, but not of Tyr59 on the alpha-cluster. These data indicate that pentamidine specifically modifies the beta-cluster, i.e., the 'basic face' of ubiquitin. Our results suggest that the biochemical mechanism of action of pentamidine may be a consequence of its dual binding to DNA and proteins.