Comparison of microfauna communities in full scale subsurface flow constructed wetlands used as secondary and tertiary treatment.

In order to evaluate the microfauna composition and distribution in two horizontal subsurface flow constructed wetlands used as secondary and tertiary treatment a full-scale wastewater treatment plant was monitored during five months. Results indicate that total microfauna abundance in the wetland treating primary influents is around five times higher than that found in the wetland treating secondary influents. Ciliated protozoa and microflagellates are the most important microfauna groups in both wetlands; microflagellates in terms of abundance and ciliates in terms of biomass. The most abundant ciliate species in the wetland treating primary influents are polysaprobic organisms as Dexiostoma campylum, Trimyema compressum, and to a lesser extend Metopus spp. On the other hand, the most important ciliate species found in the wetland treating secondary influents are mainly aerobic ciliates as Vorticella comvallaria-complex, Aspidisca cicada, Litonotus lamella and some ciliates belonging to the group of the scuticociliates and Hypotrichidae. The sort of the organic matter treated (particulated or dissolved) is at least as important as the amount of it in order to explain microfauna dynamics in constructed wetlands.

Solids accumulation in six full-scale subsurface flow constructed wetlands.

In this study, we evaluated the amount of accumulated solids in six different horizontal subsurface flow constructed wetlands (SSF CWs). We also investigated the relationship between accumulated solids and, on one hand, the wastewater quality and load and, on the other hand, the hydraulic conductivity of the granular medium. Aerobic and anaerobic biodegradability tests were also conducted on the accumulated organic matter. Experiments were carried out on full scale wastewater treatment systems consisting of SSF CWs with stabilisation ponds, which are used for the sanitation of small towns in north-eastern Spain. There were more accumulated solids near the inlet of the SSF CWs (3-57 kg dry matter (DM)/m2) than near the outlet (2-12 kgm DM/m2). Annual solids accumulation rates ranged from 0.7 to 14.3 kg DM/m2 year, and a positive relationship was observed between accumulation rates and loading rates. Most of the accumulated solids had a low level of organic matter (<20%). The results of the aerobic and anaerobic tests indicated that the accumulated organic matter was very recalcitrant and difficult to biodegrade. The hydraulic conductivity values were significantly lower near the inlet zone (0-4 m/d) than in the outlet zone (12-200 m/d). Although hydraulic conductivity tended to decrease with increasing solids accumulation, the relationship was not direct. One major conclusion of this study is that the improvement of primary treatment is necessary to avoid rapid clogging of the granular media due to solids accumulation.

Identification of a novel Ezrin-binding site in syndecan-2 cytoplasmic domain.

ERM (Ezrin/Radixin/Moesin) proteins are crosslinkers between plasma membrane proteins and the actin cytoskeleton, thereby involved in the formation of cell adhesion sites. Earlier work showed that Ezrin links syndecan-2 to the actin cytoskeleton. Here we provide evidence that the Ezrin N-terminal domain binds to the syndecan-2 cytoplasmic domain with an estimated K(D) of 0.71 microM and without the requirement of other proteins. We also studied the regions in the syndecan-2 cytoplasmic domain implicated in the binding to Ezrin. By truncating the syndecan-2 cytoplasmic domain and by oligopeptide competition assays we show that the Ezrin-binding sequence is not located in the positively charged juxtamembrane region (RMRKK), but in the neighboring sequence DEGSYD. We therefore conclude that the consensus sequence for Ezrin binding is unique among membrane proteins, suggesting a distinct regulation.

Syndecan-2 expression increases serum-withdrawal-induced apoptosis, mediated by re-distribution of Fas into lipid rafts, in stably transfected Swiss 3T3 cells.

To examine the function of syndecan-2, one of the most abundant heparan sulfate proteoglycans in fibroblasts, we obtained stably transfected Swiss 3T3 clones. We examined the effects of stable syndecan-2 overexpression on programmed cell death, finding that syndecan-2 transfected cells were more sensitive to apoptosis induced by serum-withdrawal than control cells. In addition, overexpression of syndecan-2 correlates with increased membrane levels of the Fas/CD95 receptor, suggesting that the increased serum-withdrawal apoptosis observed in Swiss 3T3 cells might be Fas receptor-dependent. Differences in Fas membrane levels between both control and syndecan-2 transfected cells result from a redistribution of the Fas receptor. Our data clearly demonstrate that increased Fas levels are primarily related to lipid rafts and that this increase is a key factor in Fas/CD95-mediated apoptosis. Moreover, disruption of lipid rafts with methyl-beta-cyclodextrin or filipin significantly reduced apoptosis in response to serum withdrawal. The differences in Fas/CD95 membrane distribution could explain why syndecan-2 transfected cells have a higher susceptibility to serum-withdrawal-induced apoptosis.

Cdk2 activation acts upstream of the mitochondrion during glucocorticoid induced thymocyte apoptosis.

Thymocytes undergo apoptosis during negative selection in vivo and following treatment with glucocorticoids or DNA-damaging drugs in vitro. The post-mitochondrial biochemical steps leading to apoptosis induced by these stimuli are well characterized, however, much less is known about the pathways connecting receptor triggering, apical caspase activation and induction of mitochondrial dysfunction. These stimuli specifically activate the kinase Cdk2 and this step is obligatory for these forms of thymocyte apoptosis. We report here that Cdk2 activation is a very early step during thymocyte apoptosis preceding apical caspase activation and phosphatidylserine exposure. Furthermore, Cdk2 activation is required for mitochondrial permeability disruption, cytochrome c release and, as a consequence, activation of the downstream caspases 9 and 3. Our data allow an integrated linear pathway regulating DNA damage and glucocorticoid-induced thymocyte apoptosis to be proposed.

Syndecan-2 expression enhances adhesion and proliferation of stably transfected Swiss 3T3 cells.

Syndecans (heparan sulfate proteoglycans) participate in cell-cell and cell-matrix adhesion and are co- and low-affinity receptors for growth factors and enzymes, respectively. We examined the influence of stable syndecan-2 expression in Swiss 3T3 cells on cell-adhesion and proliferation. Higher syndecan-2 expression changed cell morphology and increased spreading and adhesion in these cells and proliferation induced by FCS and FGF-2. This emphasizes the role of syndecan-2 in the integration of signals from soluble and insoluble factors.