Human Land use Threatens Endemic Wetland Species: The Case of Chorthippus lacustris (La Greca and Messina 1975) (Orthoptera: Acrididae) in Epirus, Greece

Chorthippus lacustris is an endemic grasshopper (Orthoptera) species in Epirus, Greece. Its population status, habitat characteristics, and relation to historical and current human land use are investigated. The species has a restricted and fragmented distribution pattern. Five locations, four within Pamvotida Lake basin and one in Lake Paramythia, cover a total of 0.12 km². It is strongly dependent on wet grasslands, flooded on a seasonal basis. The greatest population density is recorded in the site with the greatest diversity of dominant plant species. Ch. lacustris is estimated to have lost 85–99% of its habitat during the last 50 years due to wetland drainage. The main threat to the species survival is further habitat loss by urbanisation around Pamvotida Lake and by land conversion to agriculture in Paramythia Lake, even though both sites belong to the Natura 2000 network. The species status is Critically Endangered and it should be listed in Annex II of the Habitat Directive (92/43/EEC) as a priority species for conservation. Restoring wet grasslands, protecting them from further urbanisation and drainage, and monitoring species population are the main measures proposed for its conservation.     

Epidermal growth factor receptor pathway analysis identifies amphiregulin as a key factor for cisplatin resistance of human breast cancer cells

The use of platinum complexes for the therapy of breast cancer is an emerging new treatment modality. To gain insight into the mechanisms underlying cisplatin resistance in breast cancer, we used estrogen receptor-positive MCF-7 cells as a model system. We generated cisplatin-resistant MCF-7 cells and determined the functional status of epidermal growth factor receptor (EGFR), MAPK, and AKT signaling pathways by phosphoreceptor tyrosine kinase and phospho-MAPK arrays. The cisplatin-resistant MCF-7 cells are characterized by increased EGFR phosphorylation, high levels of AKT1 kinase activity, and ERK1 phosphorylation. In contrast, the JNK and p38 MAPK modules of the MAPK signaling pathway were inactive. These conditions were associated with inactivation of the p53 pathway and increased BCL-2 expression. We investigated the expression of genes encoding the ligands for the ERBB signaling cascade and found a selective up-regulation of amphiregulin expression, which occurred at later stages of cisplatin resistance development. Amphiregulin is a specific ligand of the EGFR (ERBB1) and a potent mitogen for epithelial cells. After exposure to cisplatin, the resistant MCF-7 cells secreted amphiregulin protein over extended periods of time, and knockdown of amphiregulin expression by specific short interfering RNA resulted in a nearly complete reversion of the resistant phenotype. To demonstrate the generality and importance of our findings, we examined amphiregulin expression and cisplatin resistance in a variety of human breast cancer cell lines and found a highly significant correlation. In contrast, amphiregulin levels did not significantly correlate with cisplatin resistance in a panel of lung cancer cell lines. We have thus identified a novel function of amphiregulin for cisplatin resistance in human breast cancer cells.     

Pharmacological profile of a novel H2S-releasing aspirin

The pharmacological profile of a new, safe, and effective hydrogen sulfide (H₂S)-releasing derivative of aspirin (ACS14) is described. We report the synthesis of ACS14, and of its deacetylated metabolite (ACS21), the preliminary pharmacokinetics, and its in vivo metabolism, with the H₂S plasma levels after intravenous administration in the rat. ACS14 maintains the thromboxane-suppressing activity of the parent compound, but seems to spare the gastric mucosa, by affecting redox imbalance through increased H₂S/glutathione formation, heme oxygenase-1 promoter activity, and isoprostane suppression.     

Effects of litter removal and mowing on germination and establishment of two fen-grassland species along a productivity gradient

To study the factors that limit the occurrence of species in fen grasslands, the effects of mowing and litter removal on germination, growth and establishment of two common species (Silene flos-cuculi andLotus pedunculatus) were analyzed along a productivity gradient at seven sites in Northern Germany. The sites differed in nutrient availability, vegetation composition and standing crop. In autumn 2002 a field experiment was set up at each site, including factorial combinations of two treatments (litter removal, mowing). Seeds and juvenile individuals of both species were transferred within the treatment combinations. The number of germinated and transplanted individuals was recorded during the 2003 and 2004 seasons and biomass of survived plants weighed in late summer 2004. Mowing and litter removal had positive effects on the number of germinated seeds of both species. The effect of litter removal was mostly significant at sites with a comparatively low nutrient availability and standing crop (350 g/m²), while the effect of mowing was more important at the site with the highest standing crop (1000 g/m²). In some cases the presence of litter had a positive effect on the survival of individuals, which may be caused by extremely dry weather conditions in 2003. The experiment showed that plant species reaction on mowing and litter removal differed both according to their life stage and the site conditions. These differences could be interpreted in context with Shifting Limitation Hypothesis because the abolition of seed limitation by sowing and creation of additional gaps for germination was most efficient at low productive sites, while the high standing crop reduced first of all the establishment probability, which could be increased by mowing.     

Methods to measure the intracellular concentration of unlabeled compounds within cultured cells using liquid chromatography/tandem mass spectrometry

A high-throughput and sensitive liquid chromatography/tandem mass spectrometry assay was established to detect total unlabeled hepatitis C virus inhibitor concentrations in replicon cells. The intracellular concentrations determined by this assay correlated well with concentrations obtained using radiolabeled compound. Some compounds accumulated inside the cells, with concentrations up to 300-fold higher than the input concentration. Confocal microscopic evaluation of two fluorescent-tagged inhibitors confirmed high accumulation inside the cells, sequestered inside vesicles within the cytoplasm. Incubation of cells with compound at 4 °C revealed that nonspecific binding to the outside of the cell membrane and to the cell culture plate occurred for some compounds. Therefore, the total concentration of compound extracted at 37 °C was reduced by the amount that was nonspecifically bound at 4 °C to yield the amount of compound inside the cells. A modification of the protocol was used for compounds with low intracellular concentrations in which cells were harvested with trypsin-EDTA prior to extraction. This eliminated the nonspecific binding to the cell culture plate and decreased the overall background of the assay. This assay was used to understand differences in cellular potency between compounds and the effects of serum proteins on the metabolic stability of compounds during incubation with cells.     

A highly pathogenic strain of Bacillus thuringiensis serovar kurstaki in lepidopteran pests

In order to detect and identify the most toxic Bacillus thuringiensis strains against pests, we isolated a B. thuringiensis strain (Bn1) from Balaninus nucum (Coleoptera: Curculionidae), the most damaging hazelnut pest. Bn1 was characterized via morphological, biochemical, and molecular techniques. The isolate was serotyped, and the results showed that Bn1 was the B. thuringiensis serovar, kurstaki (H3abc). The scanning electron microscopy indicated that Bn1 has crystals with cubic and bipyramidal shapes. The Polymerase Chain Reactions (PCRs) revealed the presence of the cry1 and cry2 genes. The presence of Cry1 and Cry2 proteins in the Bn1 isolate was confirmed via SDS-PAGE, at approximately 130 kDa and 65 kDa, respectively. The bioassays conducted to determine the insecticidal activity of the Bn1 isolate were conducted with four distinct insects, using spore-crystal mixtures. We noted that Bn1 has higher toxicity as compared with the standard B. thuringiensis subsp. kurstaki (HD-1). The highest observed mortality was 90% against Malacosoma neustria and Lymantria dispar larvae. Our results show that the B. thuringiensis isolate (Bn1) may prove valuable as a significant microbial control agent against lepidopteran pests.     

Relations of the morphological characteristic latent structure and body posture indicators in children aged seven to nine years

With the aim of determining the connection between the indicators of body posture and latent structure of morphological variables in children aged 7 and 8 years, first and second grade of primary school, a set of 17 morphological measures and 12 body posture indicators were longitudinally applied to a sample of 110 boys and 114 girls. The latent structure of morphological variables in both sexes was defined by three factors but at a different order of significance: in boys, the order was longitudinal dimensionality, voluminosity, mass and subcutaneous fat tissue and transverse dimensionality, whereas in girls the order was voluminosity, mass and subcutaneous fat tissue, longitudinal dimensionality and transverse dimensionality. The latent structure of torax body posture indicator was defined by two factors, the status of body posture of the rear part of the thorax, and status of the body posture of the front part of the thorax. The results obtained by canonical correlation analysis between predictive variables, morphological latent structure and criterion variables, latent structure of thorax body posture indicators with two posture indicators of the chest and one of the foot status, showed two important pairs of canonical roots on each measurement, suggesting a significant association between these two sets of parameters.     

A new model of selection in women's handball

The aim of the study was to assess the basic motor abilities that determine top performance in women's handball, and to identify test panel for primary selection at handball school. The study included 155 female attendants of the Split Handball School, mean age 12.5 years. Differences in the basic motor abilities between the subjects that developed into elite handball players after 7-year training process and those that abandoned handball for being unable to meet the competition criteria were evaluated by use of discriminative analysis. The former were found to have also been superior initially in all variables analyzed, and in arm coordination, overall body coordination, throw and jump explosive strength, arm movement frequency and repetitive trunk strength in particular. Motor superiority based on the abilities of coordination, explosive strength and speed determines performance in women's handball, qualifying these abilities as reliable selection criteria. Based on this study results, a new model of selection in women's handball, with fine arm coordination as the major limiting factor of performance, has been proposed.