The β-Hemolysin and Intracellular Survival of Streptococcus agalactiae in Human Macrophages

S. agalactiae (group B streptococci, GBS) is a major microbial pathogen in human neonates and causes invasive infections in pregnant women and immunocompromised individuals. The S. agalactiae β-hemolysin is regarded as an important virulence factor for the development of invasive disease. To examine the role of β-hemolysin in the interaction with professional phagocytes, the THP-1 monocytic cell line and human granulocytes were infected with a serotype Ia S. agalactiae wild type strain and its isogenic nonhemolytic mutant. We could show that the nonhemolytic mutants were able to survive in significantly higher numbers than the hemolytic wild type strain, in THP-1 macrophage-like cells and in assays with human granulocytes. Intracellular bacterial multiplication, however, could not be observed. The hemolytic wild type strain stimulated a significantly higher release of Tumor Necrosis Factor-α than the nonhemolytic mutant in THP-1 cells, while similar levels of the chemokine Interleukin-8 were induced. In order to investigate bacterial mediators of IL-8 release in this setting, purified cell wall preparations from both strains were tested and found to exert a potent proinflammatory stimulus on THP-1 cells. In conclusion, our results indicate that the β-hemolysin has a strong influence on the intracellular survival of S. agalactiae and that a tightly controlled regulation of β-hemolysin expression is required for the successful establishment of S. agalactiae in different host niches.     

Active transport of L-sorbose and 2-deoxy-D-galactose in Saccharomyces fragilis.

Sorbose and 2-deoxy-D-galactose are taken up in Saccharomyces fragilis by an active transport mechanism, as indicated by the energy requirement of the process and the accumulation of free sugar against the concentration gradient. There are no indications for transport-associated phosphorylation as mechanism of energy coupling with these two sugars. The measured sugar-proton cotransport and the influx inhibition by uncouplers suggest a chemiosmotic coupling mechanism. Thus there are at least two different active transport mechanisms operative in Saccharomyces fragilis: transport-associated phosphorylation in the case of 2-deoxy-D-glucose and chemiosmotic coupling in the case of sorbose and 2-deoxy-D-galactose. The differences between the two mechanisms are discussed. Uncouplers do not stimulate downhill sorbose transport in energy-depleted cells and evoke an almost complete inhibition of efflux and of exchange transport. The differences between this sugar-proton cotransport system and similar systems in bacteria and Chlorella are discussed.     

Dynamic measurement of the overall volumetric mass-transfer coefficient in air-sparged systems

The determination of the overall volumetric mass-transfer coefficient with the dynamic measurement technique involves modeling, parameter estimation, and experimental design. The combination and extension of previous efforts lead to some suggestions for improvements.     

Significance of partial pre-acidification of glucose for methanogenesis in an anaerobic digestion process

Summary The effect of partial pre-acidification of carbohydrate containing wastewaters on anaerobic digester performance was investigated. The influent was a 1% (w/v) glucose solution in a mineral salts medium imposing carbon-limited growth conditions. Up to 13% of the Chemical Oxygen Demand (COD) was added as volatile fatty acids (VFA).In all cases, addition of VFA to the glucose medium resulted in significant increases in the maximum specific COD-conversion rates of the sludge (both with respect to continuous feeding and following a shock loading), as compared with values found on digestion of glucose media alone.     

Inducible DNA polymerase I synthesis in a UV hyper-resistant mutant of Escherichia coli

A mutant of Escherichia coli which is more resistant to shortwave UV light than its wild-type parent strain and which can synthesise DNA polymerase I constitutively has been further analysed. It carries two mutational alleles which are located about 1.5 min apart and cotransducible by P1 with the argH locus. The two mutational alleles have been segregated and their analysis shows that one of them is responsible for UV hyper-resistance whereas the other mutation confers UV sensitivity. Recombinant plasmids carrying various sections of the polA regulatory region, linked to a galK gene, were introduced into the mutant strains. Analysis of galactokinase shows that the enzyme activity in the UV hyper-resistant mutant is increased. The results suggest that the synthesis of DNA polymerase I in E. coli is inducible.     

Estimation of tyrosine-40-DNA distance in the filamentous phage Pf1 by analysis of its intrinsic fluorescence properties

Iodination of the exposed Tyr-25 in the coat protein decreases the fluorescence intensity of the filamentous phage Pf1 to less than 3% of its original fluorescence. If one assumes that the total residual fluorescence originates from the non-iodinated, buried Tyr-40, one can estimate the distance between Tyr-40 and the DNA bases in Pf1 to be less than 7 A, making use of the Foerster law for fluorescence energy transfer. The result is consistent with the idea that Tyr-40-DNA interaction is responsible for the unusually large axial base separation in Pf1-DNA.