Contaminant Removal of Domestic Wastewater by Constructed Wetlands： Effects of Plant Species

A comparative study of the efficiency of contaminant removal between five emergent plant species and between vegetated and unvegetated wetlands was conducted in small-scale （2.0 m×1.0 m×0.7 m, lengthxwidthxdepth） constructed wetlands for domestic wastewater treatment in order to evaluate the decontaminated effects of different wetland plants. There was generally a significant difference in the removal of total nitrogen （TN） and total phosphorus （TP）, but no significant difference in the removal of organic matter between vegetated and unvegetated wetlands. Wetlands planted with Canna indica Linn., Pennisetum purpureum Schum., and Phragmites communis Trin. had generally higher removal rates for TN and TP than wetlands planted with other species. Plant growth and fine root （root diameter ≤ 3 mm） biomass were related to removal efficiency. Fine root biomass rather than the mass of the entire root system played an important role in wastewater treatment. Removal efficiency varied with season and plant growth. Wetlands vegetated by P. purpureum significantly outperformed wetlands with other plants in May and June, whereas wetlands vegetated by P. communis and C. indica demonstrated higher removal efficiency from August to December. These findings suggest that abundance of fine roots is an important factor to consider in selecting for highly effective wetland plants. It also suggested that a plant community consisting of multiple plant species with different seasonal growth patterns and root characteristics may be able to enhance wetland performance.     

Establishing a minimum dataset for soil quality assessment based on soil properties and land-use changes

To assess soil quality, a minimum dataset (MDS) of soil properties has to be proposed commonly through calculating the total load of each candidate soil parameter on all of the qualified principal components by use of principal component analysis (PCA) and Norm-value computation. Considering intensive land-use changes, the method introduced in this study on MDS establishment integrates the quantified contributions of land-use type and land-use duration on each soil parameter by using multivariate analysis and mean multiple comparison. In this way, a MDS maximally representing all candidates with minimal loss of the soil quality information contained by those non-MDS soil parameters is established. The MDS proposed can not only well integrate the quantified influence of land-use changes and land-use duration on soil parameters, but is also quite flexible and extendable with the potential to be extrapolated to assess soil quality in other regions. Based on two sets of soil database obtained separately in 1985 and 2004, two MDSs established are compared with each other. It is found that only quite a small change in MDS components occurs during a 20-year period. For a better assessment of soil quality, it seems necessary to examine on what kind of temporal scale and how much MDS will change for a site-specific area with intensive land-use changes.     

Hierarchical recruitment by AMPA but not staurosporine of pro-apoptotic mitochondrial signaling in cultured cortical neurons: evidence for caspase-dependent/independent cross-talk

Excitotoxicity mediated via the ( S )-α-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) subtype of receptor for l -glutamate contributes to various neuropathologies involving acute brain injury and chronic degenerative disorders. In this study, AMPA-induced neuronal injury and staurosporine (STS)-mediated apoptosis were compared in primary neuronal cultures of murine cerebral cortex by analyzing indices up- and downstream of mitochondrial activation. AMPA-mediated apoptosis involved induction of Bax, loss of mitochondrial transmembrane potential (ΔΨ_m), early release of cytochrome c (cyt c ), and more delayed release of second mitochondrial activator of caspases (SMAC), Omi, and apoptosis-inducing factor (AIF) with early calpain and minor late activation of caspase 3. STS-induced apoptosis was characterized by a number of differences, a more rapid time course, non-involvement of ΔΨ_m, and relatively early recruitment of SMAC and caspase 3. The AMPA-induced rise in intracellular calcium appeared insufficient to evoke ΔΨ_m as release of cyt c preceded mitochondrial depolarization, which was followed by the cytosolic translocation of SMAC, Omi, and AIF. Bax translocation preceded cyt c release for both stimuli inferring its involvement in apoptotic induction. Inclusion of the broad spectrum caspase inhibitor zVAD-fmk reduced the AMPA-induced release of cyt c , SMAC, and AIF, while only affecting the redistribution of Omi and AIF in the STS-treated neurons. Only AIF release was affected by a calpain inhibitor (calpastatin) which exerted relatively minor effects on the progression of cellular injury. AMPA-mediated release of apoptogenic proteins was more hierarchical relative to STS with its calpain activation and caspase-dependent AIF redistribution arguing for a model with cross-talk between caspase-dependent/independent apoptosis.     

Functional screening of salt stress-related genes from Thellungiella halophila using fission yeast system

Salinity stress is one of the most serious factors limiting plant growth and threatening global food security. Identification of salt tolerance relevant genes from halophytes can provide more insights into the determinants of salt stress tolerance in plants. In this study, functional screening of genes for the salinity tolerance in Thellungiella halophila (salt cress) was carried out using fission yeast as a host organism. A cDNA library of the salt cress, constructed in the yeast vector under the control of the inducible promoter nmt1, was transformed into the fission yeast Schizosaccharomyces pombe. Transgenic yeast cells that showed enhanced salt tolerance were selected. A total of 69 such clones were identified from 10⁶ of yeast transformants by one-time screening. Functional assignment indicates that a large percentage (81%) of these cDNAs encode for homologues of proteins that are known to have roles in plant salt tolerance, while the other portion (19%) codes for unknown proteins. Among the proteins encoded by the known functional genes, the regulatory proteins account for a relatively higher proportion. Northern blot analysis showed that the expression of some isolated genes was salt stress inducible, whereas that of the others was constitutive. These results indicate that fission yeast is a simple and efficient system for functional and large-scale identification of salt tolerance-related genes from other organisms.     

Retention of Virulence in a Viable but Nonculturable Edwardsiella tarda Isolate

Edwardsiella tarda is pathogen of fish and other animals. The aim of this study was to investigate the viable but nonculturable (VBNC) state and virulence retention of this bacterium. Edwardsiella tarda CW7 was cultured in sterilized aged seawater at 4°C. Total cell counts remained constant throughout the 28-day period by acridine orange direct counting, while plate counts declined to undetectable levels (<0.1 CFU/ml) within 28 days by plate counting. The direct viable counts, on the other hand, declined to ca. 10⁹ CFU/ml active cells and remained fairly constant at this level by direct viable counting. These results indicated that a large population of cells existed in a viable but nonculturable state. VBNC E. tarda CW7 could resuscitate in experimental chick embryos and in the presence of nutrition with a temperature upshift. The resuscitative times were 6 days and 8 days, respectively. The morphological changes of VBNC, normal, and resuscitative E. tarda CW7 cells were studied with a scanning electron microscope. The results showed that when the cells entered into the VBNC state, they gradually changed in shape from short rods to coccoid and decreased in size, but the resuscitative cells did not show any obvious differences from the normal cells. The VBNC and the resuscitative E. tarda CW7 cells were intraperitoneally inoculated into turbot separately, and the fish inoculated with the resuscitative cells died within 7 days, which suggested that VBNC E. tarda CW7 might retain pathogenicity.     

Characterization of the osmoprotectant transporter OpuC from Pseudomonas syringae and demonstration that cystathionine-beta-synthase domains are required for its osmoregulatory function

The plant pathogen Pseudomonas syringae may cope with osmotic stress on plants, in part, by importing osmoprotective compounds. In this study, we found that P. syringae pv. tomato strain DC3000 was distinct from most bacterial species in deriving greater osmoprotection from exogenous choline than from glycine betaine. This superior osmoprotection was correlated with a higher capacity for uptake of choline than for uptake of glycine betaine. Of four putative osmoregulatory ABC transporters in DC3000, one, designated OpuC, functioned as the primary or sole transporter for glycine betaine and as one of multiple transporters for choline under high osmolarity. Surprisingly, the homolog of the well-characterized ProU transporter from Escherichia coli and Salmonella enterica serovar Typhimurium did not function in osmoprotection. The P. syringae pv. tomato OpuC transporter was more closely related to the Bacillus subtilis and Listeria monocytogenes OpuC transporters than to known osmoprotectant transporters in gram-negative bacteria based on sequence similarity and genetic arrangement. The P. syringae pv. tomato OpuC transporter had a high affinity for glycine betaine, a low affinity for choline, and a broad substrate specificity that included acetylcholine, carnitine, and proline betaine. Tandem cystathionine-beta-synthase (CBS) domains in the ATP-binding component of OpuC were required for transporter function. The presence of these CBS domains was correlated with osmoregulatory function among the putative transporters examined in DC3000 and was found to be predictive of functional osmoregulatory transporters in other pseudomonads. These results provide the first functional evaluation of an osmoprotectant transporter in a Pseudomonas species and demonstrate the usefulness of the CBS domains as predictors of osmoregulatory activity.     

A role for diacylglycerol in annexin A7-mediated fusion of lung lamellar bodies

Lung surfactant secretion in alveolar type II cells occurs following lamellar body fusion with plasma membrane. Annexin A7 is a Ca2+-dependent membrane-binding protein that is postulated to promote membrane fusion during exocytosis in some cell types including type II cells. Since annexin A7 preferably binds to lamellar body membranes, we postulated that specific lipids could modify the mode of annexin A7 interaction with membranes and its membrane fusion activity. Initial studies with phospholipid vesicles containing phosphatidylserine and other lipids showed that certain lipids affected protein interaction with vesicle membranes as determined by change in protein tryptophan fluorescence, protein interaction with trans membranes, and by protein sensitivity to limited proteolysis. The presence of signaling lipids, diacylglycerol or phosphatidylinositol-4,5-bisphosphate, as minor components also modified the lipid vesicle effect on these characteristics and membrane fusion activity of annexin A7. In vitro incubation of lamellar bodies with diacylglycerol or phosphatidylinositol-4,5-bisphosphate caused their enrichment with either lipid, and increased the annexin A7 and Ca2+-mediated fusion of lamellar bodies. Treatment of isolated lung lamellar bodies with phosphatidylinositol- or phosphatidylcholine phospholipase C to increase diacylglycerol, without or with preincubation with phosphatidylinositol-4,5-bisphosphate, augmented the fusion activity of annexin A7. Thus, increased diacylglycerol in lamellar bodies following cell stimulation with secretagogues may enhance membrane fusion activity of annexin A7.