haemolysin of Escherichia coli: Comparison of pore-forming properties between chromosome and plasmid-encoded haemolysins

Abstract Lipid bilayer experiments were performed with chromosome-encoded haemolysin of Escherichia coli . The addition of the toxin to the aqueous phase bathing lipid bilayer membranes of asolectin resulted in the formation of transient ion-permeable channels with two states at small transmembrane voltages. One is prestate (single-channel conductance 40 pS in 0.15 M KCl) of the open state, which had a single-channel conductance of 420 pS in 0.15 M KCl and a mean lifetime of 30 s. Membranes formed of pure lipids were rather inactive targets for this haemolysin. Experiments with different salts suggested that the haemolysin channel was highly cation-selective at neutral pH. The mobility sequence of the cations in the channel was similar if not identical to their mobility sequence in the aqueous phase. The single-channel data were consistent with a wide, water-filled channel with an estimated minimal diameter of about 1 nm. The pore-forming properties of chromosome-encoded haemolysin were compared with those of plasmid-encoded haemolysin. Both toxins share common features, oligomerize probably to form pores in lipid bilayer membranes. Both types of haemolysin channels have similar properties but different lifetimes.     

Isolation and Biochemical Characterization of a Neural Proteoglycan Expressing the L5 Carbohydrate Epitope

The monoclonal L5 antibody reacts with an N-glycosidically linked carbohydrate structure which is present on the neural cell adhesion molecule L1, neural chondroitin sulfate proteoglycans, and other not yet identified glycosylated proteins. Using this antibody, we isolated and characterized proteoglycans from adult mouse brain and cultured astrocytes biosynthetically labeled with Na2 35SO4 and a 3H-amino acid mixture. Our data suggest that the L5 proteoglycans of both sources are identical in their biochemical properties. The apparent molecular mass of the L5 proteoglycan is approximately 500 kDa. Digestion of the iodinated L5 proteoglycan from mouse brain and of the [35S]methionine-labeled L5 proteoglycan from cultured astrocytes with proteinase-free chondroitinases ABC and AC revealed three major core proteins with apparent molecular masses of approximately 380, 360, and 260 kDa. These represent molecularly distinct protein cores.     

Expression of c-Fos in subcutaneous adipose tissue of the fetal pig

Calcium oxalate crystal growth and aggregation leads to the formation of renal calculi. It is known to be inhibited by several compounds both in vitro and in vivo conditions. The present study highlights the inhibitory potential of sodium pentosan polysulphate (SPP), a semi-synthetic glycosaminoglycan (GAG) on calcium oxalate crystal growth in vitro. Its efficacy was compared with those of known inhibitors like pyrophosphate, heparin and chondroitin-4-sulphate. Of the above compounds pyrophosphate was found to be the most potent inhibitor. Among the GAGs, SPP exhibited 80% inhibitory activity as compared to heparin. A lesser degree of inhibition was observed with chondroitin-4-sulphate.     

Isolation, characterization, and expression of a second beta-tubulin-encoding gene from Colletotrichum gloeosporioides f. sp. aeschynomene.

Colletotrichum gloeosporioides f. sp. aeschynomene is a fungal plant pathogen of Aeschynomene virginica. A beta-tubulin-encoding gene (TUB2) from this pathogen was cloned and sequenced. The deduced amino acid sequence of TUB2 had a high degree of homology to other fungal beta-tubulins. A portion of TUB2 from a benomyl-resistant C. gloeosporioides f. sp. aeschynomene mutant was also cloned and sequenced. A point mutation resulting in a glutamic acid-to-lysine substitution at amino acid 198 likely confers benomyl resistance. The mutation is relevant for use as a selectable marker in developing a gene transfer system in C. gloeosporioides f. sp. aeschynomene. Northern (RNA) hybridizations with C. gloeosporioides f. sp. aeschynomene TUB2 and another C. gloeosporioides f. sp. aeschynomene beta-tubulin-encoding gene (TUB1) as probes showed differential expression of these genes in different cell types.     

Expression of Biologically Active Basic Fibroblast Growth Factor by Genetically Modified Rat Primary Skin Fibroblasts

Abstract: Basic fibroblast growth factor (FGF-2) is normally expressed as a cell-associated protein, and accordingly it is not clear how it exerts its action on target cells in vivo. It has been proposed that cells release, by death or other mechanisms, small amounts of FGF-2 that then acts in an autocrine manner. To address the question of whether it is necessary that FGF-2 remain cell associated or needs to be secreted from cells to have biological activity, we expressed the 18-kDa form of FGF-2 in primary fibroblasts as a cell-associated (FGF-2-B) or as a secreted (FGF-2-S) protein. FGF-2 protein is detected in cell lysates and membrane fractions of both cell types, whereas it is present in significant amounts only in the conditioned medium of FGF-2-S cells. No FGF-2 is detected in control (untransfected) cells. FGF-2-S cells also grow faster than the control or FGF-2-B cells. Yet, when evaluated for their ability to promote the survival of embryonic hippocampal neurons in vitro, both the cell types are active, establishing the activity of the transgene product. We conclude that FGF-2 is active when engineered to be expressed as a cell-associated form or secreted from cells.     

Microdistribution of butterflies in a mosaic-like habitat: The role of nectar sources

The microdistribution of five butterfly species through their flying season was analyzed in a mosaic-like habitat, brought about by secondary succession In order to explain the patterns observed, activity patterns and the use and distribution of nectar sources were determined Emphasis was laid on the changing allocation of visits to flower species and changing abundances of flowers during the season The use of nectar sources was basically limited to three flower species, Centaurea scabiosa, C bracteata and Serratula tinctoria As a consequence, niche breadth values were generally low and niche overlaps generally high Some butterflies changed their patterns of flower visits during the season and therefore reduced niche overlap with the other butterfly species The microdistribution of Melanargia galathea, Lysandra condon, Ochlodes venatus and Lictoria achilleae was strongly influenced by the distribution of their preferred nectar sources as well as by areas generally rich in flowers Changing flower preferences of Melanargia galathea and Lysandra coridon males during the course of the season were also expressed by changes in the correlations between the distribution of these butterflies and their nectar plants The distribution of nectar sources was not found to be of importance for Coenonympha arcanta, a species which rarely visited flowers     

Scrotal circumference does not accurately predict degree of germinal epithelial loss or semen quality in yearling Hereford and Simmental bulls

The accuracy of scrotal circumference as a predictor of specific pathologic changes within testicular parenchyma was assessed by scoring 121 Hereford and Simmental bulls for scrotal circumference, degree of germinal epithelial loss and semen quality. Scrotal circumference was not linearly related to the degree of germinal epithelial loss or to the percentage of tubules (Grade 4+ or higher, no germinal epithelium) for either breed (P > 0.1). Thirty-two centimeters was the minimal acceptable scrotal circumference to ensure both a low prevalence of tubules with irreversible loss of germinal epithelium and acceptable semen quality. Scrotal circumference had a positive predictive value (the proportion of test-positive animals truly diseased) of 0.07 (95% confidence interval [CI] of 0 to 0.29) and a specificity (the proportion of test postive individuals that are diseased) of 0.89 (95% CI of 0.83 to 0.95) for 25% Grade 4 + tubules. For acceptable semen morphology (>/= 75% morphologically normal spermatozoa), scrotal circumference had a positive predictive value of 0.08 (95% CI of 0 to 0.34), a sensitivity (the proportion of diseased individuals testing positively) of 0.06 (95% CI of 0 to 0.24), and a specificity of 0.88 (95% CI of 0.80 to 0.95). For acceptable sperm motility (>/= 30% progressively motile spermatozoa) scrotal circumference had a positive predictive value of 0.0 (95% CI of 0 to 0.22), a sensitivity of 0.0 (95% CI of 0 to 0.15), and a specificity of 0.87 (95% CI of 0.80 to 0.94). Although scrotal circumference is widely accepted as a useful test, its accuracy should be recognized as low when 32 cm is used as the minimal acceptable scrotal circumference in predicting unacceptable testicular histology or unacceptable semen quality in a healthy population of yearling bulls.     

Dispersal failure contributes to plant losses in NW Europe

The ongoing decline of many plant species in Northwest Europe indicates that traditional conservation measures to improve the habitat quality, although useful, are not enough to halt diversity losses. Using recent databases, we show for the first time that differences between species in adaptations to various dispersal vectors, in combination with changes in the availability of these vectors, contribute significantly to explaining losses in plant diversity in Northwest Europe in the 20th century. Species with water- or fur-assisted dispersal are over-represented among declining species, while others (wind- or bird-assisted dispersal) are under-represented. Our analysis indicates that the 'colonization deficit' due to a degraded dispersal infrastructure is no less important in explaining plant diversity losses than the more commonly accepted effect of eutrophication and associated niche-based processes. Our findings call for measures that aim to restore the dispersal infrastructure across entire regions and that go beyond current conservation practices.