The study of the reaction of terminated oligomerization in the synthesis of oligo-(β1-6)-glucosamines

The applicability of terminated oligomerization to the synthesis of oligo-(β1-6)-glucosamines, fragments of the intercellular polysaccharide adhesin of staphylococci, was studied. The reactions of terminated oligomerization were carried out with mono-, di-, and trisaccharide monomers and N-protected aminopropanol; and spacered mono-and disaccharides as terminating molecules were also attempted. The primary formation of cyclic products of monomer intramolecular glycosylation was observed in almost all the reactions. Only the experiments with the monomer based on the disaccharide bromide under the conditions of the Helferich reaction led to reduced yields (30%) of the cyclic products. However, even in this case, the desired terminated oligosaccharides were generated in approximately 10% yield and mainly were the products of single glycosylation of the terminator by the monomer. These experiments allow the conclusion that, under the examined conditions, the reaction of terminated oligomerization could not result in the synthesis of oligoglucosamines with a high molecular mass.     

The function of TLR4 in interferon gamma or interleukin-13 exposed and lipopolysaccharide stimulated gingival epithelial cell cultures

Gingival epithelial cells are part of the first line of host defense against infection. Toll-like receptors (TLRs) serve important immune and nonimmune functions. We investigated how interferon gamma (INF-γ) and interleukin 13 (IL-13) are involved in the TLR4 ligand-induced regulation of interleukin-8 (IL-8) effects on gingival epithelial cells. We used immunohistochemistry to localize TLR4 in ten healthy and ten periodontitis tissue specimens. Gingival epithelial cells then were primed with Th1 cytokine (INF-γ) or Th2 cytokine (IL-13) before stimulation with Escherichia coli-derived lipopolysaccharide (LPS) and enzyme-linked immunosorbent assay (ELISA) was performed to detect the level of IL-8 secretion in cell culture supernatants. Although both healthy and periodontitis gingival tissue samples expressed TLR4, the periodontitis samples showed more intense expression on gingival epithelial cells. Gingival epithelial cell cultures were primed with either INF-γ or IL-13 before stimulation with TLR4 ligand. Supernatants from co-stimulated epithelial cells exhibited IL-8 production in opposite directions, i.e., as one stimulates the release, the other reduces the release. INF-γ significantly increased TLR4 function, whereas IL-13 significantly decreased TLR4 function, i.e., production of IL-8. Pathogen associated molecular pattern-LPS, shared by many different periodonto-pathogenic bacteria, activates the gingival epithelial cells in a TLR-dependent manner. Diminished or increased TLR function in gingival epithelial cells under the influence of different Th cell types may protect or be harmful due to the altered TLR signaling.     

Studies on the effects of low temperatures on lactic acid bacteria

Studies were conducted on different strains of L. bulgaricus, L. casei, S. thermophilus, S. lactis, and S. cremoris isolated in Bulgaria and applied as pure cultures and in combinations as starters. All the strains under investigation were found to preserve, on “freezing-thawing” their characteristic morphological and biochemical properties, regardless of the temperature and rate of cooling, but the optimum freezing temperature of the strains studied is ?196 °C (in liquid nitrogen). High cooling rates provide higher viability and activity of lactic acid bacterial cells. Lactic acid streptococci, S. lactis and S. thermophilus, are considerably more resistant than lactic acid rods, L. casei and L. bulgaricus, at all the freezing regimens tested.     

Geographic variations of seasonality and coexistence in communities: The role of diversity and climate

One of the most conspicuous and widely analyzed patterns in ecology is the latitudinal gradient in species richness. Over the 200 years since its recognition, several hypotheses have accumulated in order to account for spatial variations in diversity. Geographic variations in seasonality have been repeatedly proposed as a determinant of community richness. However, the geographic structure of community seasonality has not yet been analyzed. In the present work we evaluated three hypotheses that account for variations in the temporal structuring of communities: first, environmental seasonality determines community seasonality; second, community richness determines its degree of structuring; and third, the presence of an increase in species segregation with latitude, reflected in a pattern of species negative co‐occurrence. The hypotheses were evaluated using path analysis on 29 amphibian communities from South America, connecting latitude, environmental conditions, diversity, seasonality, and coexistence structure – nestedness and negative co‐occurrence – within communities. Latitude positively affects community seasonality through an increase in temperature seasonality, but a weak negative direct effect suggests that other variables not considered in the model – such as the strength of biotic interactions – could also be involved. Both latitude and diversity (directly and indirectly) determine an increase in negative co‐occurrence and nestedness. This suggests that groups of species that are mutually nested in time are internally segregated. Further, the strength of this structure is determined by community diversity and latitude. Temporal structuring of a community is associated with latitude and diversity, pointing to the existence of a systematic change in community organization far beyond, but probably interrelated, with the recognized latitudinal trend in richness. The available information and analysis supported the three hypotheses evaluated.     

Stathmin Potentiates Vinflunine and Inhibits Paclitaxel Activity

Cell biology and crystallographic studies have suggested a functional link between stathmin and microtubule targeting agents (MTAs). In a previous study we showed that stathmin increases vinblastine (VLB) binding to tubulin, and that conversely VLB increases stathmin binding to tubulin. This constituted the first biochemical evidence of the direct relationship between stathmin and an antimitotic drug, and revealed a new mechanism of action for VLB. The question remained if the observed interaction was specific for this drug or represented a general phenomenon for all MTAs. In the present study we investigated the binding of recombinant stathmin to purified tubulin in the presence of paclitaxel or another Vinca alkaloid, vinflunine, using Isothermal Titration Calorimetry (ITC). These experiments revealed that stathmin binding to tubulin is increased in the presence of vinflunine, whereas no signal is observed in the presence of paclitaxel. Further investigation using turbidity and co-sedimentation showed that stathmin inhibited paclitaxel microtubule-stabilizing activity. Taken together with the previous study using vinblastine, our results suggest that stathmin can be seen as a modulator of MTA activity and binding to tubulin, providing molecular explanation for multiple previous cellular and in vivo studies showing that stathmin expression level affects MTAs efficiency.     

Conjugates of cyclooligosaccharide scaffolds and carbohydrate ligands: Methods for synthesis and the interaction with lectins

The main approaches to the preparation of mono- and oligodentate glycoconjugates based on cyclodextrin and cyclooligo-β-(1 → 6)-D-glucosamine scaffolds are surveyed in the review. The data on the biological activity of the glycoconjugates are discussed exemplified in their interaction with carbohydrate-binding proteins named lectins.     

Local adaptation in populations of a Brachionus group plicatilis cryptic species inhabiting three deep crater lakes in Central Mexico

1. Salinity is a strong selective force for many aquatic organisms, affecting both ecological and evolutionary processes. Most of our knowledge on the effects of salinity on rotifers in the Brachionus plicatilis species complex is based mainly on populations from waterbodies that experience broad environmental changes both seasonally and annually. We tested the hypothesis that, despite the supposedly high potential for gene flow among rotifers inhabiting neighbouring environments, constant salinity has promoted local adaptation, genetic population divergence and even cryptic speciation in B. plicatilis complex populations from three deep maar lakes of distinct salinities [1.1, 6.5 and 9.0 g L^?1 total dissolved solids (TDS)] in Central Mexico. 2. To look for local adaptation, we performed common garden experiments to test the effect of different salinities on population density and intrinsic growth rate (r). Then, we evaluated the genetic divergence by sequencing the cytochrome c oxidase subunit I (COI) gene and performed reproductive trials to assess the potential gene flow among the three populations and with other closely related B. plicatilis complex species. 3. We confirmed that the rotifer populations have phenotypic plasticity in tolerance of salinity, but only rotifers from the least saline lake are adapted to low salinity. Among the populations, sequence divergence at COI was very low (just a single haplotype was found), suggesting a persistent founder effect from a relatively recent single colonisation event and a subsequent dispersal from one lake to the others, and a very restricted immigration rate. In the phylogenetic analysis, rotifers from this area of Mexico clustered in the same clade with the middle‐sized species Brachionus ibericus and B. sp. ‘Almenara’. Mexican rotifers showed successful recognition, copulation and formation of hybrids among them, but interpopulation breeding with the Spanish B. ibericus and B. sp. ‘Almenara’ was unsuccessful. 4. We conclude that the B. plicatilis complex populations from these three lakes belong to a new biological species not yet described (presently named B. sp. ‘Mexico’). To our knowledge, this is the first report of local adaptation of a natural B. plicatilis complex population living in freshwater conditions (1.1 g L^?1 TDS).