Fluorometric detection of enzyme activity with synthetic supramolecular pores

The reversible blockage of synthetic pores formed by rigid-rod beta barrels, either by substrates or products, was used to sense a variety of enzymatic reactions in high-throughput format with "naked-eye" fluorescent detection. Improvement of sensor sensitivity beyond three orders of magnitude by straightforward internal mutations underscores the functional plasticity of rigid-rod beta barrels. Such detectors of enzyme activity with the aforementioned characteristics are needed in areas as diverse as proteomics and environmentally benign organic synthesis.     

Was there really an Archean phosphate crisis?

During the Archean, massive amounts of iron were deposited in the form of banded iron formations. It has been suggested that sedimenting particles of ferric oxyhydroxide may have stripped dissolved phosphate from the oceans, causing a reduction in phytoplankton productivity. However, that model does not take into account the high concentration of dissolved silica that was present in seawater at that time. We show experimentally that silica effectively competes with phosphate for sorption sites on ferrihydrite particles. Furthermore, coprecipitation of silica with ferrihydrite reduces particle reactivity toward phosphate. Hence, Archean oceans probably contained considerably more phosphate than previously predicted.     

耐盐性Imt1基因表达载体的构建及其在酵母中的高效表达

 将Imt1（Inositol O-methyltransferase，肌醇甲基转移酶）构建成酵母表达载体pBD#-3，在酵母双突变体gpd#+（-）(glycerol 3-phosphate-dehydrogenase,3-磷酸甘油脱氢酶)中高效表达，使gpd#+（-）突变株对氯化钠的耐性由2.5%提高到5.0%。生化分析表明，在每mg干酵母细胞内积累高达2.2nmol的Ononitol(芒柄醇)含量。Western blot分析证明了Imt1基因的表达。这一研究为植物耐盐基因工程开辟了一条新途径。     

The formation of terrestrial food webs in glacier foreland: Evidence for the pivotal role of decomposer prey and intraguild predation

We investigated the structure of invertebrate food webs at three glacier foreland sites of an age of 2-34, ca. 60 and ca. 120 years in the European Alps at 2250-2450 m asl. The trophic structure was investigated by analyzing stable isotope ratios of ¹⁵N/¹⁴N and ¹³C/¹²C. The results suggest that the formation of terrestrial food webs during early primary succession heavily relies on prey out of the decomposer system with Collembola being most important. The diet of decomposers likely is based predominantly on allochthonous humus material blown in by wind and deposited by the retreating glacier. Irrespective of the successional stage the animal community consisted mainly of generalist predators with a number of species occurring at each of the successional stages. The results suggest that terrestrial food web formation is associated with a prolongation of food chains caused mainly by predator species switching their diet to include other predators, i.e. by intraguild predation. This suggests that generalist predators, such as cursorial spiders, carabid beetles, harvestman and centipedes, switch prey and include other predators if these are becoming more abundant, i.e. if ecosystems become more productive. Intraguild predation results in complex food webs with high linkage density which likely affects food web functioning and stability.     

Respiratory function and pulmonary lesions in pigs infected with porcine reproductive and respiratory syndrome virus

Pulmonary dysfunction was evaluated in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV, isolate VR-2332) and compared to clinical and pathological findings. Infected pigs developed fever, reduced appetite, respiratory distress and dullness at 9 days post-inoculation (dpi). Non-invasive pulmonary function tests using impulse oscillometry and rebreathing of test gases (He, CO) revealed peripheral airway obstruction, reduced lung compliance and reduced lung CO-transfer factor. PRRSV-induced pulmonary dysfunction was most marked at 9–18 dpi and was accompanied by a significantly increased respiratory rate and decreased tidal volume. Expiration was affected more than inspiration. On histopathological examination, multifocal areas of interstitial pneumonia (more severe and extensive at 10 dpi than 21 dpi) were identified as a possible structural basis for reduced lung compliance and gas exchange disturbances.     

Probing mode of action in plant cell cycle by the herbicide endothall, a protein phosphatase inhibitor

The mode of action of endothall, an herbicide which was reported to inhibit plant protein phosphatases 1 (PP1) and 2A (PP2A), was investigated. For initial characterization, a series of bioassays was used for comprehensive physiological profiling of endothall effects which suggested a phytotoxic mode of action similar to mitotic disrupter herbicides. Unlike known microtubule disrupters, endothall did not inhibit soybean tubulin polymerization in vitro. As shown in meristematic corn root tips, endothall distorted the orientation of cell division plane and microtubule spindle structures which led to cell cycle arrest in prometaphase. In tobacco BY-2 cells, malformed spindles together with prometaphase arrest of nuclei and abnormal perinuclear microtubule patterns were detected as early as 4 h of endothall treatment. These effects were also observed after treatment with other protein phosphatase inhibitors, cantharidin and okadaic acid, which phenocopied the mitotic changes described in tonneau1 (ton1) and tonneau2 (ton2) Arabidopsis mutants. These mutants are defective in TONNEAU2 (TON2) protein, a regulatory subunit of PP2A, which governs cell division plane and microtubule orientation. Therefore, PP2A/TON2 phosphatase complex is suggested to be an in planta molecular target of endothall. However, in BY-2 cells, additional effects of endothall, including inhibition of S-phase initiation and DNA synthesis, detected by 5-ethynyl-2′-deoxyuridine (EdU) incorporation, and condensed nuclei arrested in late mitosis were observed which were not reported in Arabidopsiston1 and ton2 mutants. This result indicates that two additional checkpoints in cell cycle were blocked by endothall which are probably not associated with TON2-pathway inhibition. Possibly, inhibition of PP1 and/or other PP2A protein phosphatases are involved in the regulation of these cell cycle phenomena.     

Dynamic plant uptake model applied for drip irrigation of an insecticide to pepper fruit plants

BACKGROUND: Drip application of insecticides is an effective way to deliver the chemical to the plant that avoids off‐site movement via spray drift and minimizes applicator exposure. The aim of this paper is to present a cascade model for the uptake of pesticide into plants following drip irrigation, its application for a soil‐applied insecticide and a sensitivity analysis of the model parameters. RESULTS: The model predicted the measured increase and decline of residues following two soil applications of an insecticide to peppers, with an absolute error between model and measurement ranging from 0.002 to 0.034 mg kg fw^?1. Maximum measured concentrations in pepper fruit were approximately 0.22 mg kg fw^?1. Temperature was the most sensitive component for predicting the peak and final concentration in pepper fruit, through its influence on soil and plant degradation rates. CONCLUSION: Repeated simulations of pulse inputs with the cascade model adequately describe soil pesticide applications to an actual cropped system and reasonably mimic it. The model has the potential to be used for the optimization of practical features, such as application rates and waiting times between applications and before harvest, through the integrated accounting of soil, plant and environmental influences. Copyright © 2011 Society of Chemical Industry