Changes in Hg fractionation in soil induced by willow

This study investigated the effect of willow (Salix viminalis ×  S. schwerinii) on soil characteristics, including changes in Hg fractionation in the soil solid phase, and Hg accumulation and distribution in pot-grown plants cultivated for 32 and 76 days in aged Hg-contaminated soil (30 mg Hg kg⁻¹DW). Changes in soil pH and organic carbon content as well as in Hg fractionation were monitored in both rhizospheric soil and in soil without plants. Mercury fractionation was performed by a 5-step sequential soil extraction procedure. Organic carbon content increased while pH decreased in the rhizospheric soil. Both chemically defined exchangeable Hg (0.1) and Hg bound to humic and fulvic acids (1.1) decreased in the rhizospheric soil, whereas plant accumulation of Hg increased with cultivation time. The sum of the decrease of these two soil Hg fractions after 76 days of cultivation was approximately equal to the amount of Hg accumulated in plants. Furthermore, the major Hg fractions (Hg bound to residual organic matter (53), sulphides (43), and the residual fraction (2.5)) remained stable. Neither whole plant accumulation of Hg from the soil, approximately 0.2 of total Hg in soil after 76 days cultivation, nor the fraction of total plant Hg in the shoots, which accounted for about 3 of the total plant Hg pool regardless of the cultivation time, were high. The overall results suggest that plants might be suitable for phytostabilization of aged Hg-contaminated soil, where root systems trap bioavailable Hg and help to control both leaching of Hg and re-entrainment of Hg-containing particulates from a contaminated site.     

Role of rhizosphere mechanisms in Cd uptake by various wheat cultivars

In each wheat type, cultivars have different propensities to accumulate Cd in their grains, likely depending on Cd uptake by roots and/or Cd distribution in the plant. This study investigates the processes in the root–soil interface and their role in high or low grain Cd accumulation. Twenty-four cultivars of spring bread, winter bread, durum, and spelt wheat with different grain Cd accumulation levels were investigated regarding removal of Cd from soil, pH, Cd and organic acids in root exudates, and cation-exchange capacity of roots (rootCEC). In addition, we investigated ¹⁰⁹Cd uptake from a nutrient solution resembling soil solution. The removal of Cd from the rhizosphere soil increased, likely due to increased rootCEC with increased grain Cd accumulation propensity, except in spring bread wheat. The ¹⁰⁹Cd uptake from solution did not differ between high and low grain Cd accumulators. If the soil Cd concentration was elevated, rootCEC increased, as did pH, and succinic acid levels in the exudates, while lactic and citric acid levels in root exudates decreased. This work indicates that high grain Cd accumulators take up more Cd from soil than do low accumulators. But not by a different capacity to take up Cd from soil solution. The higher rootCEC in high accumulating cultivars may influence the release of Cd from the soil particles.     

Increased Ca++ or mg++ concentration reduces relative tight-junction permeability to Na+ in the cortical thick ascending limb of Henle's loop of rabbit kidney

We have tested whether increased Ca++ and Mg++ concentrations have an effect on transepithelial voltage (PDte) and transepithelial resistance (Rte) in isolated perfused cortical thick ascending limbs (cTAL) of rabbit kidney. The divalent cations added at 2.5, 5.0 and 10.0 mmol.l-1 to the lumen or peritubular bath perfusate led to a concentration-dependent increase in Rte. The maximal response in Rte was observed between 5 and 10 mmol.l-1. No significant change in active transepithelial potential difference (PDte) was observed. The increase in Rte still occurred when the transcellular current was reduced by Ba++ (3 mmol.l-1) added to the lumen perfusate. This suggests that the increase in Rte caused by Ca++ and Mg++ is due to a modification of the paracellular shunt pathway. In the absence of active transport, i.e. when furosemide (5.10(-5) mol.l-1) was added to the lumen perfusate. Ca++ and Mg++ reduced the transepithelial diffusion potential generated by a NaCl gradient established across the epithelium, and thus produced a reduction of the relative permeability for Na+ over Cl- (PNa+/PCl-) of the paracellular shunt pathway. This indicates that divalent cations increase Rte by reducing the sodium permeability of the tight junctions. The observed Ca++ and Mg++ induced reduction of the sodium permeability of the paracellular pathway corresponds to a decrease in net Na+ reabsorption by 5-10%. Since it has been demonstrated that peptide hormones such as parathyrin (PTH) modulate divalent cation and NaCl reabsorptions, in a second series of experiments we tested the effects of PTH (2-20 USP.l-1) and dbcAMP (10(-3) mol.l-1) on PDte and Rte of isolated perfused cTAL segments of rabbit nephron. Neither Rte nor PDte were affected by PTH or dbcAMP.     

Purification and characterization of Dolichos lablab lectin

The mannose/glucose-binding Dolichos lablab lectin (designated DLL) has been purified from seeds of Dolichos lablab (hyacinth bean) to electrophoretic homogeneity by affinity chromatography on an ovalbumin- Sepharose 4B column. The purified lectin gave a single symmetric protein peak with an apparent molecular mass of 67 kDa on gel filtration chromatography, and five bands ranging from 10 kDa to 22 kDa upon SDS-PAGE. N-Terminal sequence analysis of these bands revealed subunit heterogeneity due to posttranslational proteolytic truncation at different sites mostly at the carboxyl terminus. The carbohydrate binding properties of the purified lectin were investigated by three different approaches: hemagglutination inhibition assay, quantitative precipitation inhibition assay, and ELISA. On the basis of these studies, it is concluded that the Dolichos lablab lectin has neither an extended carbohydrate combining site, nor a hydrophobic binding site adjacent to it. The carbohydrate combining site of DLL appears to most effectively accommodate a nonreducing terminal alpha-d-mannosyl unit, and to be complementary to the C-3, C-4, and C-6 equatorial hydroxyl groups of alpha-d-mannopyranosyl and alpha-d-glucopyranosyl residues. DLL strongly precipitates murine IgM but not IgG, and the recent finding that this lectin interacts specifically with NIH 3T3 fibroblasts transfected with the Flt3 tyrosine kinase receptor and preserves human cord blood stem cells and progenitors in a quiescent state for prolonged periods in culture, make this lectin a valuable tool in biomedical research.      

Nucleotide variation at the hypervariable esterase 6 isozyme locus of Drosophila simulans

Esterase 6 (Est-6/EST6) is polymorphic in both Drosophila melanogaster and D. simulans for two common allozyme forms, as well as for several other less common variants. Parallel latitudinal clines in the frequencies of the common EST6-F and EST6-S allozymes in these species have previously been interpreted in terms of a shared amino acid polymorphism that distinguishes the two variants and is subject to selection. Here we compare the sequences of four D. simulans Est-6 isolates and show that overall estimates of nucleotide heterozygosity in both coding and 5' flanking regions are more than threefold higher than those obtained previously for this gene in D. melanogaster. Nevertheless, the ratio of replacement to exon silent-site polymorphism in D. simulans is less than the ratio of replacement to silent divergence between D. simulans and D. melanogaster, which could be the result of increased efficiency of selection against replacement polymorphisms in D. simulans or to divergent selection between the two species. We also find that the amino acid polymorphisms separating EST6- F and EST6-S in D. simulans are not the same as those that separate these allozymes in D. melanogaster, implying that the shared clines do not reflect shared molecular targets for selection. All comparisons within and between the two species reveal a remarkable paucity of variation in a stretch of nearly 400 bp immediately 5' of the gene, indicative of strong selective constraint to retain essential aspects of Est-6 promoter function.      

Histone H2AX is phosphorylated at sites of retroviral DNA integration but is dispensable for postintegration repair

The histone variant H2AX is rapidly phosphorylated (denoted gammaH2AX) in large chromatin domains (foci) flanking double strand DNA (dsDNA) breaks that are produced by ionizing radiation or genotoxic agents and during V(D)J recombination. H2AX-deficient cells and mice demonstrate increased sensitivity to dsDNA break damage, indicating an active role for gammaH2AX in DNA repair; however, gammaH2AX formation is not required for V(D)J recombination. The latter finding has suggested a greater dependence on gammaH2AX for anchoring free broken ends versus ends that are held together during programmed breakage-joining reactions. Retroviral DNA integration produces a unique intermediate in which a dsDNA break in host DNA is held together by the intervening viral DNA, and such a reaction provides a useful model to distinguish gammaH2AX functions. We found that integration promotes transient formation of gammaH2AX at retroviral integration sites as detected by both immunocytological and chromatin immunoprecipitation methods. These results provide the first direct evidence for the association of newly integrated viral DNA with a protein species that is an established marker for the onset of a DNA damage response. We also show that H2AX is not required for repair of the retroviral integration intermediate as determined by stable transduction. These observations provide independent support for an anchoring model for the function of gammaH2AX in chromatin repair.