Effects of soil organic matter and nitrogen supply on competition between Festuca ovina and Deschampsia flexuosa during inland dune succession

We tested the prediction that the successional replacement of plant species during succession on inland sand dunes results from the effects of an increase in nitrogen mineralization on competitive interactions. The growth and competitive strength of Festuca ovina and Deschampsia flexuosa on soil substrates with different amounts of soil organic matter or nitrogen supply were measured. Small tillers of Festuca ovina and Deschampsia flexuosa were grown in monocultures and 1:1-mixtures on soil columns with undisturbed layers of soil organic matter from different successional age. There was (a) no visible soil organic matter, (b) a thin soil organic layer (0.5 cm) and (c) a thicker soil organic layer (6.0 cm) present on the soil columns. The species were also grown on columns with no visible soil organic matter (bare sand) with two different levels of N fertilization to mimic the increased N mineralization in the older successional stages.In monoculture, Festuca produced more biomass on the substrates with a soil organic layer compared to the unfertilized sand substrate. It also produced more biomass on sand substrates with N fertilization. Deschampsia produced more biomass in treatments with a soil organic layer compared to the bare sand treatments, but did not respond to the ammonium-nitrate addition. In competition, Festuca seemed to be the stronger competitor on the unfertilized sand substrate. Festuca was also the better competitor on the N fertilized sand treatments, while on the treatments with a soil organic layer Deschampsia was the winning species. Our results do not support the hypothesis that an increase in N supply is responsible for the replacement of Festuca by Deschampsia that concur with the accumulation of soil organic matter during succession in inland dunes.     

High-performance liquid chromatography–tandem electrospray mass spectrometry for the determination of lidocaine and its metabolites in human plasma and urine

A sensitive, selective and accurate high-performance liquid chromatographic–tandem mass spectrometric assay was developed and validated for the determination of lidocaine and its metabolites 2,6-dimethylaniline (2,6-xylidine), monoethylglycinexylidide and glycinexylidide in human plasma and urine. A simple sample preparation technique was used for plasma samples. The plasma samples were ultrafiltered after acidification with phosphoric acid and the ultrafiltrate was directly injected into the LC system. For urine samples, solid-phase extraction discs (C₁₈) were used as sample preparation. The limit of quantification (LOQ) was improved by at least 10 times compared to the methods described in the literature. The LOQ was in the range 1.6–5 nmol/l for the studied compounds in plasma samples.     

LC/CE-MS tools for the analysis of complex arabino-oligosaccharides

Recently, various branched arabino-oligosaccharides as present in a sugar beet arabinan digest were characterized using NMR. Although HPAEC often has been the method of choice to monitor the enzymatic degradation reactions of polysaccharides, it was shown that HPAEC was incapable to separate all known linear and branched arabino-oligosaccharides present. As this lack of resolution might result in an incorrect interpretation of the results, other separation techniques were explored for the separation of linear and branched arabino-oligosaccharides. The use of porous-graphitized carbon liquid chromatography with evaporative light scattering and mass detection as well as capillary electrophoresis with laser-induced fluorescence and mass detection demonstrated the superiority of both the techniques toward HPAEC by enabling the separation and unambiguous identification of almost all the linear and branched arabino-oligosaccharides available. The elution behavior of all arabino-oligosaccharides for the three tested separation techniques was correlated with their chemical structures and conclusions were drawn for the retention mechanisms of the arabino-oligosaccharides on the different chromatographic and electrophoretic systems. The combination of the elution/migration behavior on LC/CE and the MS fragmentation patterns of the arabino-oligosaccharides led to the prediction of structures for new DP6 arabino-oligosaccharides in complex enzyme digests.     

Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis

α-Galacto-oligosaccharides (α-GOS) are produced by transgalactosylation reactions of α-galactosidase (α-Gal) or by conversion of raffinose family oligosaccharides by levansucrase. Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. α-Gal of Lactobacillus reuteri formed (1?→?3)-, (1?→?4)- or (1?→?6)-linked α-GOS but exhibited a preference for formation of (1?→?6)-linkages. Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1?→?3)-, (1?→?4)- or (1?→?6)-linked hetero-oligosaccharides. By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts.     

Preselection of predatory mites to improve year-round biological control of western flower thrips in greenhouse crops

In spring and summer, two groups of natural enemies are successfully used for biological control of western flower thrips,Frankliniella occidentalis (Pergande) in greenhouses: phytoseiid mites (Amblyseius cucumeris (Oudemans) and, to a lesser extent,A. barkeri (Hughes)) and anthocorid bugs (Orius spp.). During winter, however, these predators often fail to control the pest. One likely cause for failure is the predators' tendency to enter diapause under short day conditions. In addition, eggs of predatory mites are generally susceptible to low humidity conditions, which often arise in greenhouses when outside temperatures drop below zero, or at bright, hot days in summer. In search for a thrips predator that is not hampered by these conditions, five subtropical phytoseiid species were selected which were known to feed on thrips:A. hibisci (Chant),A. degenerans Berlese,A. limonicus s.s. Garman and McGregor,A. scutalis (Athias-Henriot) andA. tularensis (Congdon). These species were compared toA. cucumeris andA. barkeri, with respect to the following features: (1) predation and oviposition rate with youngF. occidentalis larvae as prey, (2) oviposition rate on a diet of sweet pepper pollen, (3) drought tolerance spectrum of eggs, and (4) incidence of reproductive diapause under short day conditions. The results showed thatA. limonicus exhibited the highest predation and oviposition rates on a diet of thrips larvae. Moreover,A. limonicus females showed total absence of diapause under the conditions tested. A major disadvantage of this species was, however, that its eggs were most sensitive to low air humidity conditions. Least sensitive to low air humidity were eggs ofA. degenerans andA. hibisci. Females ofA. degenerans andA. hibisci also showed total absence of diapause, and intermediate rates of predation and oviposition, on both thrips larvae and pollen. In conclusion, we argue thatA. degenerans andA. hibisci are the most promising candidates for biological control ofF. occidentalis under conditions of low humidity and short day length. The success of these candidates remains to be shown in greenhouse experiments.     

Pleiotropic drug resistance and gene amplification in a SEWA mouse tumor cell line : Complex relations revealed by drug uptake data, and lipid and protein analysis

SEWA mouse lines resistant to actinomycin D (AMD) or vincristine (VCR) exhibit the pleiotropic drug resistance (PDR) phenotype, and express a low-MW protein (p21) and numerous double minutes (DM). In drug uptake studies these lines were compared with the non-resistant parental line and with a methotrexate (MTX)-resistant line, not exhibiting PDR. On treatment with labelled AMD or VCR the two PDR lines displayed a highly reduced intracellular content of drug, whereas uptake of MTX was unchanged. Uptake of AMD was shown to be temperature-dependent. The MTX-resistant line did not exhibit any significant change in AMD or VCR uptake. Other workers have emphasized the role of a high-MW glycoprotein in the development of PDR. A search for a similar glycoprotein in our cells was unsuccessful. Since all indications point to membrane factors being important in the development of PDR, the lines were also subjected to lipid analysis. Compared with control cells distinct differences were detected in the lipid composition of all resistant lines (including the MTX-resistant line). In the course of our experiments, the DM in our most AMD-resistant line were replaced by two homogeneously staining regions (HSR). Simultaneously, the overproduction of p21 ceased, but the PDR phenotype persisted. This event tends to implicate a minor role for the p21 protein in PDR, but similar transitions from DM to HSR in other AMD-resistant SEWA lines were not accompanied by a decrease in p21 over-production. Our data point to a complex genetic control of multi-drug resistance.     

Advanced adenoma diagnosis with FDG PET in a visibly normal mucosa: a case report

A 56 year old aromatherapist presented with advanced renal failure following chronic coal tar creosote vapour inhalation, and a chronic tubulo-interstitial nephritis was identified on renal biopsy. Following dialysis dependence occult inhalation continued, resulting in seizures, ataxia, cognitive impairment and marked generalised cerebral atrophy. We describe for the first time a case of creosote abuse by chronic vapour inhalation, resulting in significant morbidity. Use of the polycyclic aromatic hydrocarbon-containing wood preservative coal tar creosote is restricted by many countries due to concerns over environmental contamination and carcinogenicity. This case demonstrates additional toxicities not previously reported with coal tar creosote, and emphasizes the health risks of polycyclic aromatic hydrocarbon exposure.     

c-Jun N-terminal kinase (JNK) signaling: Recent advances and challenges

c-Jun N-terminal kinases (JNKs), first characterized as stress-activated members of the mitogen-activated protein kinase (MAPK) family, have become a focus of inhibitor screening strategies following studies that have shown their critical roles in the development of a number of diseases, such as diabetes, neurodegeneration and liver disease. We discuss recent advances in the discovery and development of ATP-competitive and ATP-noncompetitive JNK inhibitors. Because understanding the modes of actions of these inhibitors and improving their properties will rely on a better understanding of JNK structure, JNK catalytic mechanisms and substrates, recent advances in these areas of JNK biochemistry are also considered. In addition, the use of JNK gene knockout animals is continuing to reveal in vivo functions for these kinases, with tissue-specific roles now being dissected with tissue-specific knockouts. These latest advances highlight the many challenges now faced, particularly in the directed targeting of the JNK isoforms in specific tissues.