Effect of GABA,a Bacterial Metabolite,on Pseudomonas fluorescens Surface Properties and Cytotoxicity

Different bacterial species and, particularly Pseudomonas fluorescens, can produce gamma-aminobutyric acid (GABA) and express GABA-binding proteins. In this study, we investigated the effect of GABA on the virulence and biofilm formation activity of different strains of P. fluorescens. Exposure of a psychotropic strain of P. fluorescens (MF37) to GABA (10⁻⁵ M) increased its necrotic-like activity on eukaryotic (glial) cells, but reduced its apoptotic effect. Conversely, muscimol and bicuculline, the selective agonist and antagonist of eukaryote GABA_A receptors, respectively, were ineffective. P. fluorescens MF37 did not produce biosurfactants, and its caseinase, esterase, amylase, hemolytic activity or pyoverdine productions were unchanged. In contrast, the effect of GABA was associated to rearrangements of the lipopolysaccharide (LPS) structure, particularly in the lipid A region. The surface hydrophobicity of MF37 was marginally modified, and GABA reduced its biofilm formation activity on PVC, but not on glass, although the initial adhesion was increased. Five other P. fluorescens strains were studied, and only one, MFP05, a strain isolated from human skin, showed structural differences of biofilm maturation after exposure to GABA. These results reveal that GABA can regulate the LPS structure and cytotoxicity of P. fluorescens, but that this property is specific to some strains.     

The Periplasmic Oxidoreductase DsbA Is Required for Virulence of the Phytopathogen Dickeya solani

In bacteria, the DsbA oxidoreductase is a crucial factor responsible for the introduction of disulfide bonds to extracytoplasmic proteins, which include important virulence factors. A lack of proper disulfide bonds frequently leads to instability and/or loss of protein function; therefore, improper disulfide bonding may lead to avirulent phenotypes. The importance of the DsbA function in phytopathogens has not been extensively studied yet. Dickeya solani is a bacterium from the Soft Rot Pectobacteriaceae family which is responsible for very high economic losses mainly in potato. In this work, we constructed a D. solani dsbA mutant and demonstrated that a lack of DsbA caused a loss of virulence. The mutant bacteria showed lower activities of secreted virulence determinants and were unable to develop disease symptoms in a potato plant. The SWATH-MS-based proteomic analysis revealed that the dsbA mutation led to multifaceted effects in the D. solani cells, including not only lower levels of secreted virulence factors, but also the induction of stress responses. Finally, the outer membrane barrier seemed to be disturbed by the mutation. Our results clearly demonstrate that the function played by the DsbA oxidoreductase is crucial for D. solani virulence, and a lack of DsbA significantly disturbs cellular physiology.     

Conversion of Nuclear Waste to Molten Glass: Cold‐Cap Reactions in Crucible Tests

The feed‐to‐glass conversion, which comprises complex chemical reactions and phase transitions, occurs in the cold cap during nuclear waste vitrification. To investigate the conversion process, we analyzed heat‐treated samples of a simulated high‐level waste feed using X‐ray diffraction, electron probe microanalysis, leaching tests, and residual anion analysis. Feed dehydration, gas evolution, and borate phase formation occurred at temperatures below 700°C before the emerging glass‐forming melt was completely connected. Above 700°C, intermediate aluminosilicate phases and quartz particles gradually dissolved in the continuous borosilicate melt, which expanded with transient foam. Knowledge of the chemistry and physics of feed‐to‐glass conversion will help us control the conversion path by changing the melter feed makeup to maximize the glass production rate.     

Isotopic composition of soil,vegetation or cattle hair no suitable indicator of nitrogen balances in permanent pasture

Stable isotope signatures of cattle tail switch hair or meadow vegetation have been found to be related to nitrogen (N) surpluses of whole farms and of meadows, respectively. Permanent pastures are more patchy in terms of nutrient inputs and outputs and N balances for the whole plot do not necessarily give correct impressions of losses. We here investigated correlations between isotopic signatures and N balances calculated for different spatial and temporal scales in permanent pastures. N concentrations and δ¹⁵N values of cattle tail switch hair, vegetation and soil samples were measured in an experiment with different grazing intensities started in 2002. Results were compared to soil surface balances calculated for the whole plot or for plot areas affected by either dung, urine, grazing without excreta input, or the pasture area without dung pats. There were no significant correlations between plant or cattle hair isotopic signatures and any of the balances. N fixation probably influenced vegetation signatures, making the isotopic values less dependent on soil and more on atmospheric N. The cattle preferred short mixed vegetation with more legume biomass, which also influenced the ¹⁵N values of their hair. The ¹⁵N signatures of soil samples were the best indicators of partial N balances in these heterogeneous pastures, probably because soil values are most directly influenced by N inputs and outputs. Still, soil signatures only explained between 15 and 35% of the variation in balance results. Thus, none of the tested parameters can be used as a reliable indicator of N balance results in this heterogeneous system with small differences in budgets among treatments and potentially small plot-scale N losses.     

Effect of processing conditions on the yield and failure response of an aliphatic polyketone terpolymer

The effect of processing conditions on the yield and failure behavior of an aliphatic polyketone terpolymer was studied. Testing and characterization were performed on samples that were extruded in the form of hollow cylinders. We performed the extrusion process at different shear rates and at different cooling rates to assess the effect that process conditions had on the polymer properties. We performed biaxial testing on the samples to characterize the failure envelopes, including the ductile–brittle transition condition for each process condition. The effect of shear rate was negligible, whereas the cooling rate significantly affected the failure behavior. To explain these differences in behavior, we performed characterization via differential scanning calorimetry, wide‐angle X‐ray diffraction, attenuated total reflection IR spectroscopy, dynamic mechanical thermal analysis, scanning electron microscopy, and residual stress measurements. A broad glass transition was found for all samples at temperatures higher than previously reported for this material. Alteration of the processing conditions did not influence the crystalline phase (percentage crystallinity, crystalline orientation, crystallite size, etc.). A change in spherulitic structure was also observed with altered cooling rate and is suggested to have contributed to the change in failure behavior. Residual stresses also affected the behavior of all samples. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 318–334, 2002; DOI 10.1002/app.10334     

Heterodera schachtii Nematodes Interfere with Aphid-Plant Relations on Brassica oleracea

Aboveground and belowground herbivore species modify plant defense responses differently. Simultaneous attack can lead to non-additive effects on primary and secondary metabolite composition in roots and shoots. We previously found that aphid (Brevicoryne brassicae) population growth on Brassica oleracea was reduced on plants that were infested with nematodes (Heterodera schachtii) prior (4 weeks) to aphid infestation. Here, we examined how infection with root-feeding nematodes affected primary and secondary metabolites in the host plant and whether this could explain the increase in aphid doubling time from 3.8 to 6.7 days. We hypothesized that the effects of herbivores on plant metabolites would depend on the presence of the other herbivore and that nematode-induced changes in primary metabolites would correlate with reduced aphid performance. Total glucosinolate concentration in the leaves was not affected by nematode presence, but the composition of glucosinolates shifted, as gluconapin concentrations were reduced, while gluconapoleiferin concentrations increased in plants exposed to nematodes. Aphid presence increased 4-methoxyglucobrassicin concentrations in leaves, which correlated positively with the number of aphids per plant. Nematodes decreased amino acid and sugar concentrations in the phloem. Aphid population doubling time correlated negatively with amino acids and glucosinolate levels in leaves, whereas these correlations were non-significant when nematodes were present. In conclusion, the effects of an herbivore on plant metabolites were independent of the presence of another herbivore. Nematode presence reduced aphid population growth and disturbed feeding relations between plants and aphids.