Influence of Iron on Production of the Antibacterial Compound Tropodithietic Acid and Its Noninhibitory Analog in <i>Phaeobacter inhibens</i>

Tropodithietic acid (TDA) is an antibacterial compound produced by some Phaeobacter and Ruegeria spp. of the Roseobacter clade. TDA production is studied in marine broth or agar since antibacterial activity in other media is not observed. The purpose of this study was to determine how TDA production is influenced by substrate components. High concentrations of ferric citrate, as present in marine broth, or other iron sources were required for production of antibacterially active TDA. However, when supernatants of noninhibitory, low-iron cultures of Phaeobacter inhibens were acidified, antibacterial activity was detected in a bioassay. The absence of TDA in nonacidified cultures and the presence of TDA in acidified cultures were verified by liquid chromatography–high-resolution mass spectrometry. A noninhibitory TDA analog (pre-TDA) was produced by P. inhibens, Ruegeria mobilis F1926, and Phaeobacter sp. strain 27-4 under low-iron concentrations and was instantaneously converted to TDA when pH was lowered. Production of TDA in the presence of Fe(3+) coincides with formation of a dark brown substance, which could be precipitated by acid addition. From this brown pigment TDA could be liberated slowly with aqueous ammonia, and both direct-infusion mass spectrometry and elemental analysis indicated a [Fe(III)(TDA)(2)](x) complex. The pigment could also be produced by precipitation of pure TDA with FeCl(3). Our results raise questions about how biologically active TDA is produced in natural marine settings where iron is typically limited and whether the affinity of TDA to iron points to a physiological or ecological function of TDA other than as an antibacterial compound

其一

The filamentous fungus Stachybotrys chartarum is known for its toxic metabolites and has been associated with serious health problems, including mycotoxicosis, among occupants of contaminated buildings. Here, we present results from a case study, where an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for known and tentatively identified compounds characterized via UHPLC-quadruple time-of-flight (QTOF) screening of fungal culture extracts, wall scrapings and reference standards. The UHPLC-MS/MS method was able to identify 12 Stachybotrys metabolites, of which four could be quantified based on authentic standards and a further six estimated based on similarity to authentic standards. Samples collected from walls contaminated by S. chartarum in a water-damaged building showed that the two known chemotypes, S and A, coexisted. More importantly, a link between mycotoxin concentrations found on contaminated surfaces and in settled dust was made. One dust sample, collected from a water-damaged room, contained 10 pg/cm(2) macrocyclic trichothecenes (roridin E). For the first time, more than one spirocyclic drimane was detected in dust. Spirocyclic drimanes were detected in all 11 analysed dust samples and in total amounted to 600 pg/cm(2) in the water-damaged room and 340 pg/cm(2) in rooms adjacent to the water-damaged area. Their wide distribution in detectable amounts in dust suggested they could be good candidates for exposure biomarkers. [Figure: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-016-9649-y) contains supplementary material, which is available to authorized users

Abundance and diversity of bees visiting flowering pennycress, a new oilseed crop in the midwestern USA

Oilseed pennycress (Thlaspi arvense) is a new, autumn-sown, “cash cover crop” for the Midwestern USA and elsewhere. Anthesis occurs in early spring when few other plants bloom, and its flowers attract early-emerging bees. However, the taxonomic composition of these bees was unknown. Consequently, we systematically captured and identified the genera and species of bees visiting pennycress flowers throughout anthesis at five site-years: two in Illinois and three in Minnesota. A cumulative total of 28 bee species were found across site-years. The most common genera were Andrena (10 species), Lasioglossum (12 species), and Halictus (2 species). Rarer genera were Apis, Ceratina, Hylaeus, and Nomada. Bee abundance and diversity were related closely and in a negative exponential manner with percent land area devoted to annual cropping. The inclusion of new early flowering crops, such as pennycress, may enhance bee abundance and diversity, especially if even small areas of uncropped land are nearby

Flies associated with floral canopies of the new oilseed crop, pennycress, in the Midwestern U.S.A.

Flies are frequent visitors to flowers of many species of plants within the mustard family (Brassicaceae). They derive nutrition from these flowers, and some fly species are pollinators. Field pennycress (Thlaspi arvense) is a mustard species that is being developed as a new “cash cover crop,” i.e., an autumn-sown cover crop whose oil-rich seeds can be harvested profitably in spring. Although pennycress is largely wind- and self-pollinated, its flowers also attract insect visitors. However, the extent of visitation to pennycress flowers by flies remains largely unknown, especially the identities of those flies. Thus, we examined flies associated with pennycress flowering canopies at five site-years in Illinois and Minnesota. The number of fly species averaged 16 per site-year. Hover flies (Syrphidae) were common visitors to pennycress flowering canopies, representing 24% of all Diptera collected. Toxomerus marginatus (margined calligrapher), whose larvae are aphid predators, was especially abundant within this family. However, the most common flies detected were Delia spp. (Anthomyiidae, root maggot flies), which averaged 51% of all flies collected. Adults of these flies are known pollinators, but their larvae also are pests that can damage seedlings of common summer crops. Although seedling damage to plants that are double- or relay-cropped (i.e., inter-seeded in spring) with pennycress has not been observed yet, close observation of this insect group and its effects may be needed if pennycress is widely sown in the future as a cash cover crop

Dimerisation induced formation of the active site and the identification of three metal sites in EAL-phosphodiesterases

The bacterial second messenger cyclic di-3′,5′-guanosine monophosphate (c-di-GMP) is a key regulator of bacterial motility and virulence. As high levels of c-di-GMP are associated with the biofilm lifestyle, c-di-GMP hydrolysing phosphodiesterases (PDEs) have been identified as key targets to aid development of novel strategies to treat chronic infection by exploiting biofilm dispersal. We have studied the EAL signature motif-containing phosphodiesterase domains from the Pseudomonas aeruginosa proteins PA3825 (PA3825EAL) and PA1727 (MucREAL). Different dimerisation interfaces allow us to identify interface independent principles of enzyme regulation. Unlike previously characterised two-metal binding EAL-phosphodiesterases, PA3825EAL in complex with pGpG provides a model for a third metal site. The third metal is positioned to stabilise the negative charge of the 5′-phosphate, and thus three metals could be required for catalysis in analogy to other nucleases. This newly uncovered variation in metal coordination may provide a further level of bacterial PDE regulation

Chemical Behavior and Reaction Kinetics of Sulfur and Nitrogen Half-Mustard and Iprit Carbonate Analogues

Sulfur and nitrogen mustards are very toxic, yet versatile organic molecules with numerous applications. Herein, we report on a synthesis of a new class of green compounds, i.e., half-mustard and iprit carbonates, that result in new, unexplored, and safe molecules. Their chemical behavior with several nucleophiles and their reaction kinetics have been investigated

Integrative Analysis of the Caenorhabditis elegans Genome by the modENCODE Project

We systematically generated large-scale data sets to improve genome annotation for the nematode Caenorhabditis elegans, a key model organism. These data sets include transcriptome profiling across a developmental time course, genome-wide identification of transcription factor-binding sites, and maps of chromatin organization. From this, we created more complete and accurate gene models, including alternative splice forms and candidate noncoding RNAs. We constructed hierarchical networks of transcription factor-binding and microRNA interactions and discovered chromosomal locations bound by an unusually large number of transcription factors. Different patterns of chromatin composition and histone modification were revealed between chromosome arms and centers, with similarly prominent differences between autosomes and the X chromosome. Integrating data types, we built statistical models relating chromatin, transcription factor binding, and gene expression. Overall, our analyses ascribed putative functions to most of the conserved genome