Development of an enzyme-linked glycosorbent method to monitor the inhibition of sialic acid-dependent Helicobacter pylori adhesion

Horseradish peroxidase conjugation with fetuin, which expresses sialic acid-dependent binding specificity to Helicobacter pylori, was used to develop an enzyme-linked glycosorbent method. This method yielded results that were consistent with those from a hemagglutination assay using a microscope and allowed the quantitative analysis of inhibitors of sialic acid-dependent Helicobacter pylori adhesion to host cells. The results of inhibitor screening with carbohydrates, including commercially available polysaccharides and extracted from various sources, displayed not only the relative inhibition potencies among carbohydrates, but also their respective concentration-dependencies.     

Development of enzyme-immunoassays based on egg yolk antibodies for the detection of mycotoxins

Enzyme-linked immunosorbent assays (ELISA) proved to be a fast and simple method for the detection of mycotoxins and other undesired contaminants in food and feed. The present study is focused on the optimisation and exploitation of the egg yolk antibody technology in order to develop competitive ELISAs for the detection of mycotoxins in cereals. Due to its importance as one of the most relevant Fusarium mycotoxins, the trichothecene deoxynivalenol (DON) was selected as representative. Chickens were immunised with different protein conjugates performing varying booster intervals. The antibodies were isolated by the poly(ethylene glycol) precipitation method according to Polson. By use of these antibodies an indirect competitive ELISA was developed for the detection of DON. First investigations of naturally contaminated wheat samples showed a good correspondence with results obtained by GC-ECD when calibration in blank wheat extracts was performed.     

Rheological properties of polysaccharides produced by a Zoogloea sp.

A newly isolated marine bacterium, identified as Zoogloea sp., produced two different polysaccharides: one was water-soluble and the other was cell-bound. Both had non-Newtonian, pseudoplastic fluid behaviour and the solutions had low activation energies. The solutions of these polysaccharides showed rheological behaviour over a wide range of pH (2–12) and temperature (20–80 °C), and compatibility with NaCl.     

Aerobic reduction of manganese oxide by Salmonella sp. strain MR4

A new isolate of Salmonella, strain MR4, reduced Mn(IV)O₂ at 2.3 mM under aerobic conditions by about 83% over 24 h. Direct contact of cells to MnO₂ was not necessary as the cell-free spent medium produced a similar amount of Mn(II). Pyruvate (1.6 mM) and oxalate (0.8 mM) were identified in the culture medium and presumed to have a role in Mn(II) production in this microorganism.     

Identification of ARAP3, a novel PI3K effector regulating both Arf and Rho GTPases, by selective capture on phosphoinositide affinity matrices.

We show that matrices carrying the tethered homologs of natural phosphoinositides can be used to capture and display multiple phosphoinositide binding proteins in cell and tissue extracts. We present the mass spectrometric identification of over 20 proteins isolated by this method, mostly from leukocyte extracts: they include known and novel proteins with established phosphoinositide binding domains and also known proteins with surprising and unusual phosphoinositide binding properties. One of the novel PtdIns(3,4,5)P3 binding proteins, ARAP3, has an unusual domain structure, including five predicted PH domains. We show that it is a specific PtdIns(3,4,5)P3/PtdIns(3,4)P2-stimulated Arf6 GAP both in vitro and in vivo, and both its Arf GAP and Rho GAP domains cooperate in mediating PI3K-dependent rearrangements in the cell cytoskeleton and cell shape.     

Expression of the tobacco mosaic virus movement protein alters starch accumulation inNicotiana benthamiana

Nicotiana benthamiana plants were transformed with the movement protein (MP) gene of tobacco mosaic virus (TMV), usingAgrobacterium-mediated transformation. Plants regenerated from the transformed cells accumulated 30-kDa MP and complemented the activity of TMV MP when infected with chimeric TMVs containing defective MR These transgenic plants displayed stunting, pale-green leaves, and starch accumulations, indicating that TMV MP altered the carbon partitioning for leaves involved in TMV cell-to-cell movement.     

Residual effects of 3,4-dichloroaniline on offspring born to Ceriodaphnia cf. dubia exposed for multiple generations

One of the acknowledged limitations ofconventional toxicity tests is their inabilityto evaluate the impact of toxicants onsubsequent generations. Given their relativelyshort lifespan, cladocerans in the field may beexposed to toxicants for several generations.However, it is unclear what effect such anexposure regime could have on the cladoceranfitness after removal of the toxicant. Thispaper aimed to determine the offspring fitnessof juveniles produced by adult Ceriodaphnia cf. dubia exposed toresidual levels of 3,4-dichloroaniline overfour generations. Mass cultures of Ceriodaphnia cf. dubia were maintainedfor several generations in variousconcentrations of 3,4-dichoroaniline (0, 2.5,5, 10, 15 and 20 µg/L). The mass cultureswere re-established every generation using4th brood neonates <24 h old. Eachgeneration, 4th brood neonates <24 h oldwere also transferred individually intotoxicant-free water and examined until theproduction of their 3rd brood. F1offspring of mothers exposed to 15 and20 µg/L 3,4-dichloroaniline showedsignificantly (p < 0.05) reducedreproduction compared to the controls. Nosignificant (p > 0.05) changes inreproduction due to 3,4-dichloroaniline wereobserved for the F2 and F3 offspring. However,F4 offspring of mothers exposed to all3,4-dichloroaniline concentrations showedsignificantly (p < 0.05) increasedreproduction compared to the controls. Possibleexplanations for the varying influence ofmaternal 3,4-dichloroaniline exposure on thereproductive output of the offspring arediscussed.     

Phylogenetic relationship of Alexandrium tamiyavanichii (Dinophyceae) to other Alexandrium species based on ribosomal RNA gene sequences

The phylogenetic relationship of the thecate PSP-toxin producing dinoflagellate Alexandrium tamiyavanichii Balech to other species of Alexandrium was studied based on nucleotide sequences of the ITS1, ITS2, 5.8S, 18S and 28S subunits of the ribosomal RNA gene. These are the first such sequences available for A. tamiyavanichii, which is one of the producers of paralytic shellfish poisoning toxins in tropical waters. Based on the nucleotide sequences of the 28S, 18S and 5.8S subunits of the rRNA gene, A. tamiyavanichii grouped together with A. tamarense, A. catenella and A. fundyense. More interestingly, A. tamiyavanichii was most closely affiliated to A. tamarense isolates from Thailand. This result reaffirmed conclusions from previous studies that, for the A. tamarense/fundyense/catenella species complex, geographical origin rather than morphology seems to determine genetic relatedness. Results of this study also suggest that A. tamiyavanichii most probably belongs to the same species complex. Ribosomal RNA gene sequences do not separate the PSP toxin producing from the non-producing species of Alexandrium.     

Changes in the Glutamate Release and Uptake of Cerebellar Cells in Perinatally Nicotine-Exposed Rat Pups

Cerebellar granule and glial cells were cultured from 7 day-old rat pups after pre- and post-natal nicotine treatment. Ten days later, the basal release of glutamate in the granule cells prepared from the pre- and post-natally nicotine-exposed pups was higher and lower than the controls, respectively. The N-methyl-D-aspartate-induced release of glutamate was higher in the granule cells of post-natal nicotine exposed rats. However, the nicotine-induced glutamate release was either unchanged or was lower in the granule cells of all nicotine-treated pups. The basal glutamate uptake was higher in the glial cells from those exposed pre-natally and lower in the continuously nicotine-exposed pups. The sensitivities of L-trans-pyrrolidine-2,4-dicarboxylic acid on glutamate uptake were higher in all nicotine treated groups. There was a higher number of specific [³H]dizocilpine binding sites in the pre- or continuously nicotine-exposed group. These results suggest that the cerebellar cell properties are altered after perinatal nicotine exposure and that the development of an excitatory amino acid system might be affected differently depending on the nicotine exposure time.