Isolation and characterization of mutations in the gene encoding an endogenous Bacillus subtilis beta-galactosidase and its regulator.

We have isolated mutations that appear to inactivate the gene (lacA) encoding an endogenous beta-galactosidase activity in Bacillus subtilis and in a closely linked negative regulatory element (lacR). Both genes map to the hisA-thrA region. The lacA mutations may help to avoid some of the problems arising from the use of the Escherichia coli lacZ gene as a reporter gene in B. subtilis.     

An Evaluation of Habitat Rehabilitation on Coastal Dune Forests in Northern KwaZulu-Natal, South Africa

The rehabilitation program conducted by Richards Bay Minerals (RBM) of areas exposed to opencast surface mining of sand dunes north of Richards Bay (28°43'S, 32°12'E) on the coast of northern KwaZulu-Natal Province commenced 16 years before this study and has resulted in the development of a series of known-aged stands of vegetation. By assuming that these spatially separated stands develop along a similar pathway over time, instantaneous sampling should reveal successional or other changes usually associated with aging and should provide an opportunity to evaluate the success of rehabilitation. We compare relative densities of pioneer and secondary species, species richness, and a similarity index of the herbaceous layer, tree, beetle, millipede, bird, and small-mammal communities of rehabilitating areas of known age with those of 30-year-old unmined forests and unmined forests of unknown age adjacent to the rehabilitating area. Species richness for all but the mammalian taxa increased with increasing age of rehabilitating stands. For all taxa but the mammals and herbaceous layer, the unmined stands harbored more species than the mined rehabilitating stands. The relative densities of pioneer species of all the taxa decreased with an increase in the age of rehabilitating stands, whereas those of the secondary species increased with an increase in habitat age. Similarity between unmined stands and rehabilitating stands of different ages increased with increasing regeneration age of rehabilitating stands, suggesting that rehabilitating communities, in terms of species composition and relative densities, are developing towards the status of unmined communities. Rehabilitation based on RBM's management program of limited interference is occurring and may result in the reestablishment of a coastal dune forest ecosystem. But rehabilitation resulting from succession depends on the availability of species sources from which colonization can take place. In the Richards Bay mining operation the present mining path is laid out so that such refuges are present.     

Bone morphogenetic protein-4 enhances vascular endothelial growth factor secretion by human retinal pigment epithelial cells

Retinal pigment epithelial (RPE) cells secrete vascular endothelial growth factor (VEGF), a cytokine known to promote angiogenesis. Results from RNase protection assays (RPAs) show that RPE from non-diabetic human donors and from adult retinal pigment epithelium-19 (ARPE-19) cells expressed significant bone morphogenetic protein-4 (BMP-4) message. In addition, ARPE-19 cells cultured in high glucose (25 mM), compared to those in physiological glucose (5.5 mM) released significantly more BMP-4 into the conditioned media (CM). However, the effect of BMP-4 on the release of VEGF by ARPE-19 cells has not been studied. Accordingly, ARPE-19 cells were treated with BMP-4 to determine VEGF secretion. BMP-4 and VEGF levels in the CM and cell lysates were measured by enzyme-linked immunosorbent assay (ELISA). Cells treated with exogenous BMP-4 had higher VEGF in the CM and this treatment effect was dose- and time-dependent, while cell lysates had low levels of VEGF. Addition of cycloheximide (CHX) or actinomycin-D (ACT) significantly reduced VEGF secretion from cells treated with BMP-4, suggesting that the BMP-4-induced secretion of VEGF requires new RNA and protein synthesis. Our results suggest that BMP-4 may play a role in the regulation of ocular angiogenesis associated with diabetic retinopathy (DR) by stimulating VEGF release from RPE cells.     

Succession and soil development in man-made coastal ecosystems at the Baltic Sea

By the creation of Køge Bay Seaside Park in 1978 the opportunity was given to study the establishment of vegetation and primary succession in a man-made coastal area. As the soil consists of marine material from the Baltic Sea, no organic matter or seedbank was present. The first steps of primary succession were followed in 1980, when the initial inventories of flora immigration and soil development were carried out in coastal grasslands and plantings. The surveys were repeated in 1992 and 1993. Except for the planted woody species and a few sown grasses, all other species of plants have reached the area through natural dispersal of diaspores. The total number of species in the permanent plots has increased from 26 in 1980 to 91 in 1993. The results indicate that this number will continue to increase in the coming years until a certain level. Then it will probably decrease as a result of competition from woody species, unless the vegetation is kept in a steady state by disturbances or management. Today the area is very far from the initial situation, and the off-shore barrier has changed towards a landscape dominated by small groves and grasslands of an urban common type.     

Investigation into the intracellular fates,speciation and mode of action of selenium-containing neuroprotective agents using XAS and XFM

Background

A variety of selenium compounds have been observed to provide protection against oxidative stress, presumably by mimicking the mechanism of action of the glutathione peroxidases. However, the selenium chemistry that underpins the action of these compounds has not been unequivocally established.

Diversity of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Large-Subunit Genes from Groundwater and Aquifer Microorganisms

To test our hypothesis that microbial autotrophic CO2 fixation plays an important role in subsurface systems of two large groundwater remediation projects, several anaerobic/microaerobic aquifer and groundwater samples were taken and used to investigate the distribution and phylogenetic diversity of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) large-subunit genes. Two primer sets were designed for amplifying partial-subunit genes of RubisCO forms I and II from the DNA, directly extracted from the samples. PCR products were used to construct five clone libraries with putative RubisCO form I sequences, and two libraries of DNA amplified by form II primers. Selected clones were screened for variation by restriction fragment length polymorphism analysis, and a total of 28 clone inserts were sequenced and further analyzed. The phylogenies constructed from amino acid sequences derived from the partial RubisCO large-subunit sequences showed a distinct pattern. Diverse sequences affiliated to the cluster of green-like type IA RubisCO sequences were found, representing various obligate and facultative chemolithoautotrophic Proteobacteria, whereas type II RubisCO sequences detected were most closely related to those of thiobacilli species. An isolate obtained from aquifer enrichment culture, which has been provisionally named Halothiobacillus sp. RA13 on the basis of its 16S rDNA sequence, was found to contain both types of RubisCO genes, i.e., forms I and II. Physiological and ecological considerations are discussed in the context of additional microbial data and physicochemical properties.     

Spectral effects of the pupil in fly photoreceptors

Summary Photoreceptors of flies contain pigment granules which upon illumination of the receptors migrate towards the rhabdomere and act as a longitudinal pupil. Data in the literature concerning the effect of the pupil on the spectral sensitivity are contradictory. Therefore spectral sensitivity ofMusca photoreceptors upon light adaptation was reinvestigated.The change in spectral sensitivity of fly photoreceptors upon light adaptation as measured by Hardie (1979) was confirmed. Taking into account waveguide optics this change was explained from absorbance spectra of pupillary granules, measured by microspectrophotometry in squash preparations. Furthermore the pupil absorbance spectrum determined in vivo (Stavenga et al. 1973) was interpreted. The absence of a change in spectral sensitivity upon light adaptation measured by pupillary reflexion (Bernard and Stavenga 1979) is explained by a local-triggering of the pupil.     

Cloning and expression of a cDNA encoding betanidin 5-O-glucosyltransferase, a betanidin- and flavonoid-specific enzyme with high homology to inducible glucosyltransferases from the Solanaceae

Root NO3- uptake and expression of two root NO3- transporter genes (Nrt2;1 and Nrt1) were investigated in response to changes in the N- or C-status of hydroponically grown Arabidopsis thaliana plants. Expression of Nrt2;1 is up-regulated by NO3 - starvation in wild-type plants and by N-limitation in a nitrate reductase (NR) deficient mutant transferred to NO3- as sole N source. These observations show that expression of Nrt2;1 is under feedback repression by N-metabolites resulting from NO3- reduction. Expression of Nrt1 is not subject to such a repression. However, Nrt1 is over-expressed in the NR mutant even under N-sufficient conditions (growth on NH4NO3 medium), suggesting that expression of this gene is affected by the presence of active NR, but not by N-status of the plant. Root 15NO3- influx is markedly increased in the NR mutant as compared to the wild-type. Nevertheless, both genotypes have similar net 15NO3- uptake rates due to a much larger 14NO3- efflux in the mutant than in the wild-type. Expressions of Nrt2;1 and Nrt1 are diurnally regulated in photosynthetically active A. thaliana plants. Both increase during the light period and decrease in the first hours of the dark period. Sucrose supply prevents the inhibition of Nrt2;1 and Nrt1 expressions in the dark. In all conditions investigated, Nrt2;1 expression is strongly correlated with root 15NO3- influx at 0.2 mM external concentration. In contrast, changes in the Nrt1 mRNA level are not always associated with similar changes in the activities of high- or low-affinity NO3- transport systems.     

The effect of soil nutrient availability on retranslocation of Ca,Mg and K from senescing sapwood in Atlantic white cedar

Nutrient resorption from senescing tissues increases plant nutrient-use efficiency, and may be an adaptation to nutrient limitation. In some tree species, retranslocation of nutrients from sapwood during heartwood formation is a comparable process. We measured Ca, Mg and K concentrations in Atlantic white cedar (Chamaecyparis thyoides) stemwood samples taken from two swamps in the northeastern United States and compared them to soil mineral nutrient availability at each site. We found that Ca, Mg and K concentrations were 60–700% higher in sapwood than in the immediately adjacent heartwood, indicating retranslocation of these nutrients from senescing sapwood. Sapwood nutrient concentrations were similar between the two sites. However, nutrient concentrations in the heartwood differed significantly between the sites, as did the relative degree of Ca and Mg retranslocation from senescing sapwood. We found these differences between sites to be inversely related to significant differences in exchangeable Ca, Mg and K as well as Al concentrations in the soil. These findings suggest that the degree of nutrient retranslocation from senescing sapwood may be influenced by soil nutrient availability. This revised version was published online in July 2006 with corrections to the Cover Date.