Flash chromatography on cartridges for the separation of plant extracts: Rules for the selection of chromatographic conditions and comparison with medium pressure liquid chromatography

Empirical rules for the selection of chromatographic conditions on flash chromatography cartridges were developed, with an emphasis on gradient mode. These rules were then tested with separation of extracts from important medicinal plants including Curcuma xanthorrhiza, Piper nigrum and Salvia milthiorrhiza. Sepacore® cartridges enabled a good separation of compounds with a broad range of polarity, as typically found in plant extracts. The chromatographic resolution remained, however, lower than that achieved on classical columns packed with material of smaller particle size. For poorly soluble extracts, solid introduction gave better results than liquid injection.     

Effect of thinning on anatomical adaptations of Norway spruce needles

Conifers and other trees are constantly adapting to changes in light conditions, water/nutrient supply and temperatures by physiological and morphological modifications of their foliage. However, the relationship between physiological processes and anatomical characteristics of foliage has been little explored in trees. In this study we evaluated needle structure and function in Norway spruce families exposed to different light conditions and transpiration regimes. We compared needle characteristics of sun-exposed and shaded current-year needles in a control plot and a thinned plot with 50% reduction in stand density. Whole-tree transpiration rates remained similar across plots, but increased transpiration of lower branches after thinning implies that sun-exposed needles in the thinned plot were subjected to higher water stress than sun-exposed needles in the control plot. In general, morphological and anatomical needle parameters increased with increasing tree height and light intensity. Needle width, needle cross-section area, needle stele area and needle flatness (the ratio of needle thickness to needle width) differed most between the upper and lower canopy. The parameters that were most sensitive to the altered needle water status of the upper canopy after thinning were needle thickness, needle flatness and percentage of stele area in needle area. These results show that studies comparing needle structure or function between tree species should consider not only tree height and light gradients, but also needle water status. Unaccounted for differences in needle water status may have contributed to the variable relationship between needle structure and irradiance that has been observed among conifers.     

What factors influence the stem taper of <Emphasis Type="Italic">Eucalyptus</Emphasis>: growth,environmental conditions,or genetics?

• Introduction   

Stem taper equations have been widely used for volume estimation to varied top diameter limits or for biomass calculations. However, their main drawback is that specific calibration is often necessary for each species or clone, and accounting for genetic and environmental effects is often a challenge.     

Dentate gyrus NMDA receptors mediate rapid pattern separation in the hippocampal network

Forming distinct representations of multiple contexts, places, and episodes is a crucial function of the hippocampus. The dentate gyrus subregion has been suggested to fulfill this role. We have tested this hypothesis by generating and analyzing a mouse strain that lacks the gene encoding the essential subunit of the N-methyl-d-aspartate (NMDA) receptor NR1, specifically in dentate gyrus granule cells. The mutant mice performed normally in contextual fear conditioning, but were impaired in the ability to distinguish two similar contexts. A significant reduction in the context-specific modulation of firing rate was observed in the CA3 pyramidal cells when the mutant mice were transferred from one context to another. These results provide evidence that NMDA receptors in the granule cells of the dentate gyrus play a crucial role in the process of pattern separation.     

Social-ethical issues concerning the control strategy of animal diseases in the European Union: A survey

In 2004 a survey was conducted in the member states of the European Union designed to gain greater insight into the views on control strategies for foot and mouth disease, classical swine fever, and avian influenza with respect to the epidemiological, economic and social-ethical consequences of each of these animal diseases. This article presents the results of the social-ethical survey. A selection of stakeholders from each member state was asked to prioritize issues for the prevention and control of these diseases. A majority of stakeholders chose preventive measures as the preferred issue. An analysis was done to determine whether there were differences in views expressed by stakeholders from member states with a history of recent epidemics and ones without such a history, and whether there were regional differences. There were no differences between member states with or without a history of recent epidemics. There were indeed regional differences between the priority orders from Northern and Southern Europe on the one hand, and from Eastern Europe on the other. Nina E. Cohen is a biologist and is a researcher at the Wageningen University. She is specialized in societal and ethical issues in human–animal relationships. Her current research is focused on the social-ethical issues concerning the prevention and control of foot and mouth disease, classical swine fever and avian influenza. Marcel A.P.M. van Asseldonk has studied animal science. Currently he works at the Institute for Risk Management in Agriculture (IRMA) of the Wageningen University. He is specialized in the design and pricing of insurance policies and animal health funds for the main livestock epidemics. Elsbeth N. Stassen is a veterinarian and professor of Animals and Society at the Wageningen University. Elsbeth Stassen is specialized in animal health, animal welfare and human–animal relationships. She was a member of a governmental welfare committee during the avian influenza epidemic in the Netherlands in 2003.     

Mass spectrometry of intact V-type ATPases reveals bound lipids and the effects of nucleotide binding

The ability of electrospray to propel large viruses into a mass spectrometer is established and is rationalized by analogy to the atmospheric transmission of the common cold. Much less clear is the fate of membrane-embedded molecular machines in the gas phase. Here we show that rotary adenosine triphosphatases (ATPases)/synthases from Thermus thermophilus and Enterococcus hirae can be maintained intact with membrane and soluble subunit interactions preserved in vacuum. Mass spectra reveal subunit stoichiometries and the identity of tightly bound lipids within the membrane rotors. Moreover, subcomplexes formed in solution and gas phases reveal the regulatory effects of nucleotide binding on both ATP hydrolysis and proton translocation. Consequently, we can link specific lipid and nucleotide binding with distinct regulatory roles.     

Glutamatergic and dopaminergic neurons mediate anxiogenic and anxiolytic effects of CRHR1

The corticotropin-releasing hormone receptor 1 (CRHR1) critically controls behavioral adaptation to stress and is causally linked to emotional disorders. Using neurochemical and genetic tools, we determined that CRHR1 is expressed in forebrain glutamatergic and γ-aminobutyric acid-containing (GABAergic) neurons as well as in midbrain dopaminergic neurons. Via specific CRHR1 deletions in glutamatergic, GABAergic, dopaminergic, and serotonergic cells, we found that the lack of CRHR1 in forebrain glutamatergic circuits reduces anxiety and impairs neurotransmission in the amygdala and hippocampus. Selective deletion of CRHR1 in midbrain dopaminergic neurons increases anxiety-like behavior and reduces dopamine release in the prefrontal cortex. These results define a bidirectional model for the role of CRHR1 in anxiety and suggest that an imbalance between CRHR1-controlled anxiogenic glutamatergic and anxiolytic dopaminergic systems might lead to emotional disorders.     

Atmospheric carbon injection linked to end-Triassic mass extinction

The end-Triassic mass extinction (~201.4 million years ago), marked by terrestrial ecosystem turnover and up to ~50% loss in marine biodiversity, has been attributed to intensified volcanic activity during the break-up of Pangaea. Here, we present compound-specific carbon-isotope data of long-chain n-alkanes derived from waxes of land plants, showing a ~8.5 per mil negative excursion, coincident with the extinction interval. These data indicate strong carbon-13 depletion of the end-Triassic atmosphere, within only 10,000 to 20,000 years. The magnitude and rate of this carbon-cycle disruption can be explained by the injection of at least ~12 × 10(3) gigatons of isotopically depleted carbon as methane into the atmosphere. Concurrent vegetation changes reflect strong warming and an enhanced hydrological cycle. Hence, end-Triassic events are robustly linked to methane-derived massive carbon release and associated climate change.     

Differences in growth and nutrient uptake from a coarse-soil substrate by ectomycorrhizal- and fungicide-treated Picea abies seedlings

In order to quantify the importance of ectomycorrhizal fungi on nutrient uptake from the coarse-soil fraction of a haplic Cambisol (alumic), a microcosm study that allowed for nutrient budgets was designed. Ectomycorrhizal- and fungicide-treated spruce seedlings were grown on isolated and cleaned gneiss fragments (6.3 mm > Ø > 2 mm) from 90 cm soil depth. The substrate was the only source of Ca, K and Mg. Ectomycorrhizal seedlings showed no signs of nutrient deficiencies and biomass increased significantly compared to initial seedling biomass. Ectomycorrhizal seedlings seemed well adapted to survive on the coarse-soil substrate and acquired Ca, K and Mg from the coarse-soil substrate. Gneiss fragments of the ectomycorrhizal treatment were covered by fine roots and ectomycorrhizal hyphae, as observed microscopically. Fungicide-treated seedlings?? root development was retarded, and shoot biomass only increased from stored nutrient reserves of the seedlings. The suppression of EcM colonization by the fungicide Topas^® apparently caused a root growth inhibiting effect. Furthermore, the extinction of mycorrhiza initiated an increased nitrification and acidification and a consequent nutrient cation release triggered by nitrate in the drainage.     

Activity-guided identification of (S)-malic acid 1-O-D-glucopyranoside (morelid) and gamma-aminobutyric acid as contributors to umami taste and mouth-drying oral sensation of morel mushrooms (Morchella deliciosa Fr.)

Although morel mushrooms are widely used as tasty ingredients in savory dishes, knowledge of the key compounds evoking their attractive taste is still very fragmentary. In the present study, taste activity-guided fractionation of an aqueous morel extract by means of the recently developed taste dilution analysis (TDA) enabled the localization of several umami-like-tasting fractions as well as a fraction imparting an intense mouth-drying sensation to the oral cavity. Hydrophilic interaction liquid chromatography (HILIC), LC-MS, and amino acid analysis led to the successful identification of gamma-aminobutyric acid as the chemical inducer of the mouth-drying and mouth-coating oral sensations imparted by the morel extract. Besides the well-known umami-like taste contributors L-glutamic acid, L-aspartic acid, and succinic acid, an additional HILIC fraction was isolated and evaluated as tasting umami-like. LC-MS and NMR studies revealed that this fraction consisted of a mixture of (S)-malic acid 1-O-alpha-D-glucopyranoside and (S)-malic acid 1-O-beta-D-glucopyranoside, the structure of which could be successfully confirmed by independent synthesis. To the best of our knowledge, this morel-derived glycoside, which we named (S)-morelid, has previously not been reported in any food products. Sensory analysis of aqueous solutions of the compounds identified revealed threshold concentrations of 0.02 mmol/L for the mouth-drying effect of gamma-aminobutyric acid and 6.0 mmol/L for the umami-like, slightly sour taste of (S)-morelid.