Serodiagnosis and monitoring of contagious bovine pleuropneumonia (CBPP) with an indirect ELISA based on the specific lipoprotein LppQ of Mycoplasma mycoides subsp. mycoides SC.

An indirect ELISA, based on the specific and strongly antigenic recombinant peptide of the N'-terminal half of the lipoprotein LppQ from Mycoplasma mycoides subsp. mycoides small colony type (SC) was developed for the detection of antibodies to M. mycoides subsp. mycoides SC. It was evaluated for its suitability for serodiagnosis and monitoring of contagious bovine pleuropneumonia (CBPP). The recombinant peptide containing poly-histidine residue tails was expressed in Escherichia coli and subsequently purified by Ni(2+) chelate affinity chromatography to be used as antigen to coat microtiter ELISA plates. The specificity of the antigen was tested against rabbit hyperimmune sera directed against related Mycoplasmas of the M. mycoides cluster and with sera from cattle that were either free of CBPP, but suffered from other mycoplasmal infections such as M. bovis, or showed cross-reactions in the complement fixation test. The sensitivity of the ELISA was assessed with sera from artificially infected animals and with sera from cattle originating from areas where CBPP was endemic at the time of blood sampling. The study revealed that the ELISA was both specific and sensitive for CBPP positive bovine sera and was shown also to be robust to harsh climatic conditions.     

Analysis of non-porcine isolates of Actinobacillus suis.

Twenty-four Actinobacillus suis isolates obtained from several species of non-porcine mammals were compared to the representative porcine strains, ATCC 15557 (serotype O1) and H89-1173 (serotype O2), by O serotyping, DNA fingerprinting, PCR amplification of apxICA, apxIICA and apxIIICA toxin genes and by rrs (16S rRNA) gene sequencing. Only two strains, both equine, reacted with O1 antiserum while two others, one canine and the other feline, reacted with O2 antiserum. One equine strain reacted weakly with both antisera. No amplification of apx genes was found with the non-porcine O1 or the "not O1/O2" strains but amplification of the apxICA and apxIICA genes was observed with the two O2 strains. In addition, these two O2 strains had both BamHI and BglII fingerprints that were very similar to the porcine O2 reference strain, H89-1173 and rrs gene sequences that were identical to the A. suis reference strain ATCC 15557. Taken together, these data suggest that although many non-porcine A. suis isolates are not A. suis (sensu stricto), some isolates are genotypically as well as phenotypically similar to A. suis.     

Effect of fertilization on respiration from different sources in a sandy soil of an agricultural long-term experiment

Annual changes in stocks of soil organic carbon may be detected by measurement of heterotrophic respiration, but field studies of heterotrophic respiration in long-term fertilization experiments on sandy soils are scarce. Our objectives were to: (1)investigate the influence of fertilizer type on mineralization of soil organic carbon and crop residue, and (2) show how fertilization treatments affect the annual C balance (net ecosystem carbon balance, NECB; negative values indicate a CO₂-source) in the sandy soil of the Darmstadt experiment. Treatments were long-term mineral fertilization with cereal straw incorporation (MSI) and application of rotted farmyard manure (FYM), both treatments receiving 14 g N m^?2 year^?1. This study used δ¹³C natural abundance after introduction of a C₄ crop to distinguish between different sources of respiration. Mineralization derived from C₃ sources was similar for MSI and FYM treatments (～270 g C m^?2 year^?1). The rate of residue mineralization in MSI treatments was higher, resulting in a mineralization of 49 and 37% of initial residue C in the soil of MSI and FYM treatments, respectively. The NECB (g C m^?2 year^?1) indicated the MSI treatment (approximately ?190) as a stronger source compared with the FYM treatment (～?30).     

Local knowledge and farmer perceptions of bean diseases in the central African highlands

Central African highland farmers' perceptions of common bean disease were investigated using both phytopathology and anthropological techniques. Farmers rarely mentioned diseases as production constraints in formal questionnaires. More participatory research showed farmers often related disease symptoms to the effects of rain and soil depletion for fungal diseases, or to varietal traits for bean common mosaic virus. Rain or moisture is divided into numerous forms through which it can damage plants, both physically and through putrefaction. Most conditions associated with putrefaction appear to be linked to pathogens. Farmers have an understanding of plant health closely related to their concept of human health. In plants, this understanding is based on the prior state of plant health. Conceptually, local disease management strategies are based on prevention by managing the conditions that promote good plant health rather than by treating disease symptoms. Intervention strategies that build on local knowledge are encouraged.     

In vitro observation of dynamic ordering processes in the extracellular matrix of living, adherent cells

Collecting information at the interface between living cells and artificial substrates is exceedingly difficult. The extracellular matrix (ECM) mediates all cell-substrate interactions, and its ordered, fibrillar constituents are organized with nanometer precision. The proceedings at this interface are highly dynamic and delicate. In order to understand factors governing biocompatibility or its counterpart antifouling, it is necessary to probe this interface without disrupting labels or fixation and with sufficient temporal resolution. Here the authors combine nonlinear optical spectroscopy (sum-frequency-generation) and microscopy (second-harmonic-generation), fluorescence microscopy, and quartz crystal microgravimetry with dissipation monitoring in a strategy to elucidate molecular ordering processes in the ECM of living cells. Artificially (fibronectin and collagen I) and naturally ordered ECM fibrils (zebrafish, Danio rerio) were subjected to nonlinear optical analysis and were found to be clearly distinguishable from the background signals of diffusive proteins in the ECM. The initial steps of fibril deposition and ordering were observed in vitro as early as 1 h after cell seeding. The ability to follow the first steps of cell-substrate interactions in spite of the low amount of material present at this interface is expected to prove useful for the assessment of biomedical and environmental interfaces.     

Marine Derived Hamacanthins as Lead for the Development of Novel PDGFRβ Protein Kinase Inhibitors

In this study, we report on pyrazin-2(1H)-ones as lead for the development of potent adenosine triphosphate (ATP) competitive protein kinase inhibitors with implications as anti-cancer drugs. Initially, we identified the pyrazin-2(1H)-one scaffold from hamacanthins (deep sea marine sponge alkaloids) by Molecular Modeling studies as core binding motif in the ATP pocket of receptor tyrosine kinases (RTK), which are validated drug targets for the treatment of various neoplastic diseases. Structure-based design studies on a human RTK member PDGFR (platelet-derived growth factor receptor) suggested a straight forward lead optimization strategy. Accordingly, we focused on a Medicinal Chemistry project to develop pyrazin-2(1H)-ones as optimized PDGFR binders. In order to reveal Structure-Activity-Relationships (SAR), we established a flexible synthetic route via microwave mediated ring closure to asymmetric 3,5-substituted pyrazin-2(1H)-ones and produced a set of novel compounds. Herein, we identified highly potent PDGFR binders with IC₅₀ values in an enzymatic assay below µM range, and possessing significant activity against PDGFR dependent cancer cells. Thus, marine hamacanthin-derived pyrazin-2(1H)-ones showing interesting properties as lead for their further development towards potent PDGFR-inhibitors.     

Effect of irrigation on near-infrared (NIR) based prediction of mango maturity

This study investigated the effect of irrigation on the ability of near-infrared (NIR) measurements to predict maturity stage of mango. Fruits from irrigated and non-irrigated trees were sampled on six dates during the final three weeks of development and subjected to NIR and conventional laboratory analyses. Spectral assessment showed differentiation between irrigated and non-irrigated fruits on earlier dates, which was not evident later on. NIR measurements of irrigated samples correlated well to dry matter, this was not the case for non-irrigated samples (r = 0.80 compared to 0.57, respectively), while the reverse was true for acidity with r increasing from 0.55 to 0.85 between irrigated and non-irrigated. Second derivative spectra of all samples best correlated with acid content (r = 0.73). Although dry matter was previously proposed as the most suitable parameter for NIR calibration, this study suggests acidity might be an appropriate harvesting index when considering irrigation effects. Interestingly, NIR technology has been found to adequately predict acidity in other fruits, but results of this study require additional investigation.