Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for regioselective chlorination

Chlorinated natural products include vancomycin and cryptophycin A. Their biosynthesis involves regioselective chlorination by flavin-dependent halogenases. We report the structural characterization of tryptophan 7-halogenase (PrnA), which regioselectively chlorinates tryptophan. Tryptophan and flavin adenine dinucleotide (FAD) are separated by a 10 angstrom-long tunnel and bound by distinct enzyme modules. The FAD module is conserved in halogenases and is related to flavin-dependent monooxygenases. On the basis of biochemical studies, crystal structures, and by analogy with monooxygenases, we predict that FADH2 reacts with O2 to make peroxyflavin, which is decomposed by Cl-. The resulting HOCl is guided through the tunnel to tryptophan, where it is activated to participate in electrophilic aromatic substitution.     

COX-2-derived prostacyclin confers atheroprotection on female mice

Female gender affords relative protection from cardiovascular disease until the menopause. We report that estrogen acts on estrogen receptor subtype alpha to up-regulate the production of atheroprotective prostacyclin, PGI2, by activation of cyclooxygenase 2 (COX-2). This mechanism restrained both oxidant stress and platelet activation that contribute to atherogenesis in female mice. Deletion of the PGI2 receptor removed the atheroprotective effect of estrogen in ovariectomized female mice. This suggests that chronic treatment of patients with selective inhibitors of COX-2 could undermine protection from cardiovascular disease in premenopausal females.     

Large-scale copy number polymorphism in the human genome

The extent to which large duplications and deletions contribute to human genetic variation and diversity is unknown. Here, we show that large-scale copy number polymorphisms (CNPs) (about 100 kilobases and greater) contribute substantially to genomic variation between normal humans. Representational oligonucleotide microarray analysis of 20 individuals revealed a total of 221 copy number differences representing 76 unique CNPs. On average, individuals differed by 11 CNPs, and the average length of a CNP interval was 465 kilobases. We observed copy number variation of 70 different genes within CNP intervals, including genes involved in neurological function, regulation of cell growth, regulation of metabolism, and several genes known to be associated with disease.     

DNA: a programmable force sensor

Direct quantification of biomolecular interaction by single-molecule force spectroscopy has evolved into a powerful tool for materials and life sciences. We introduce an approach in which the unbinding forces required to break intermolecular bonds are measured in a differential format by comparison with a known reference bond (here, a short DNA duplex). In addition to a marked increase in sensitivity and force resolution, which enabled us to resolve single-base pair mismatches, this concept allows for highly specific parallel assays. This option was exploited to overcome cross-reactions of antibodies in a protein biochip application.     

Modulation of oscillatory neuronal synchronization by selective visual attention

In crowded visual scenes, attention is needed to select relevant stimuli. To study the underlying mechanisms, we recorded neurons in cortical area V4 while macaque monkeys attended to behaviorally relevant stimuli and ignored distracters. Neurons activated by the attended stimulus showed increased gamma-frequency (35 to 90 hertz) synchronization but reduced low-frequency (<17 hertz) synchronization compared with neurons at nearby V4 sites activated by distracters. Because postsynaptic integration times are short, these localized changes in synchronization may serve to amplify behaviorally relevant signals in the cortex.     

Comparative anatomic and morphometric examination of the interosseous muscle,sesamoid ligaments and flexor tendons of the fetlock in South American camelids

This study presents a comparative morphological and morphometrical examination of the interosseous muscle (IOM), digital flexor tendons and associated structures of all four species of South American Camelids (SACs): Lama (Lama glama), alpaca (Vicugna pacos), guanaco (Lama guanicoe) and vicuna (Vicugna vicugna). Our findings show that the lamoid interosseous muscle and digital flexor tendons are very similar between species and differences, which have been reported previously, are possibly due to diverging examiner interpretation. Lumbricalis muscles were mostly found as single muscles in this study. The definition of the terminal and dorsal branches of the interosseous muscle is critically revised. The analysis of morphometric data indicates that the size of the individual has a substantial influence on the cross-sectional area (CSA) of the examined structures. Metacarpal/metatarsal bone width is a main predictor of CSA, while the factor species only shows significance in specific sample points. For lama and alpaca, no differences in CSA in relation to species as factor were found. The IOM shows the largest CSAs of the three examined structures in all species, which implies an important load-bearing function. Considerable individual variation remains unexplained by the mentioned factors. This lowers the prospect of CSA reaching diagnostic importance in SAC fetlock pathology.     

The need for new concepts in risk management of sediments historical developments, future perspectives and new approaches

Being part of a highly dynamic system, contaminated sediments are especially in need of an integrated management approach. Due to change in importance from source to diffuse pollution and the variety of chemical substances in the environ-ment, different scientific fields need to cooperate and incorpo-rate their data in a common risk assessment scheme. Public perception of risk that is associated with sediments and with chemical data is low while the acceptance of ecotoxico-logical data with decision makers is often missing. A growing demand of the public to be involved in decision processes and informed about environmental problems demands a change of methods and concepts in the future. Necessity of an integration of risk assessment and management procedures has been suggested in order to increase the efficiency of the process and the early involvement of public concern. As the confidence in experts’ opinions decreases, a strong need for communication with and transparency for all involved parties arises.     

Identification of the astringent taste compounds in black tea infusions by combining instrumental analysis and human bioresponse

Application of taste dilution analyses on freshly prepared black tea infusions revealed neither the high molecular weight thearubigen-like polyphenols nor the catechins and theaflavins, but a series of 14 flavon-3-ol glycosides as the main contributors to the astringent taste perceived upon black tea consumption. Among these glycosides, the apigenin-8-C-[alpha-l-rhamnopyranosyl-(1-->2)-O-beta-d-glucopyranoside] was identified for the first time in tea infusions. Depending on the structure, the flavon-3-ol glycosides were found to induce a velvety and mouth-coating sensation at very low threshold concentrations, which were far below those of catechins or theaflavins; for example, the threshold of 0.001 micromol/L found for quercetin-3-O-[alpha-l-rhamnopyranosyl-(1-->6)-O-beta-d-glucopyranoside] is 190000, or 16000 times below the threshold determined for epigallocatechin gallate or theaflavin, respectively. Moreover, structure/activity considerations revealed that, besides the type of flavon-3-ol aglycon, the type and the sequence of the individual monosaccharides in the glycosidic chain are key drivers for astringency perception of flavon-3-ol glycosides.     

Increasing species and trophic diversity of mesofauna affects fungal biomass,mesofauna community structure and organic matter decomposition processes

A laboratory mesocosm experiment was set up to study the effects of five mesofauna model communities on litter fungal biomass (ergosterol content) and litter decomposition parameters (litter mass remaining, N concentration) for 4 months. The five treatments were: (1) no soil mesofauna, (2) the collembolan Folsomia fimetaria, (3) an assemblage of the collembolan species Isotomurus prasinus, Hypogastrura assimilis, F. fimetaria, Mesaphorura macrochaeta and Protaphorura armata, (4) the same collembolan assemblage plus the enchytraeid Enchytraeus crypticus, and (5) the six species plus the predaceous mite Hypoaspis aculeifer. After 30, 60, 90 and 120 days of incubation, six mesocosms of each treatment were sacrificed and biomass, species richness and dominance were measured. Changes occurred during the study with an increase in total detritivore biomass and a decrease in species richness with strong dominance by a few species in multispecies mesocosms. A vertical stratification of species was also observed in the mesocosms. Litter mass loss and N mineralisation were reduced with the introduction of fauna and showed the largest effects in the multispecies detritivore treatments. Ergosterol production was only temporary reduced in the multispecies treatments reflecting an effect of mesofauna on fungal biomass related to diversity rather than the biomass of mesofauna. An inhibitory predator effect was observed on functional decomposition parameters and indicated top-down control of decomposition via detritivorous mesofauna.