The extensive bloom of alternate-stage Prymnesium polylepis (Haptophyta) in the Baltic Sea during autumn–spring 2007–2008

During autumn 2007, an unusual increase in an algal species belonging to the order Prymnesiales was observed throughout the Baltic Sea Proper during routine national monitoring. Electron microscopical examination of the blooming species showed two types of flat scales – small and large – that resembled those of the alternate stage of Prymnesium polylepis. No spine-bearing scales were found. The 18S rDNA sequence data (n?=?20, c. 1500?bp) verified the species identification as P. polylepis. There was up to 0.5% (7?bp) variability in the P. polylepis partial 18?S rDNA sequences from the Baltic Sea. These environmental sequences differed by 0–0.35% (0–4?bp) from cultured P. polylepis (isolate UIO036), and by 1.0–3.7% from other available Prymnesium sequences. The number of cells assumed to be P. polylepis began to increase in October 2007 coincidently with significantly calm and dry weather, and at their maximum the cells accounted for over 80% of the total phytoplankton biovolume in December–January. During February–April 2008, 95% of the Prymnesiales cells were in the size class of P. polylepis (>6?µm). The species attained bloom concentrations (>1?×?10⁶?cells?l^–1) from March to May 2008. The species was observed throughout the Baltic Sea, except the Bothnian Bay, Gulf of Riga and the Kattegat. No toxic effects of the bloom were observed.     

Aminopeptidase N (APN)/CD13-dependent CXCR4 downregulation is associated with diminished cell migration,proliferation and invasion

Aminopeptidase N (APN/CD13) is a 150 kDa membrane-bound ubiquitously expressed protease with a broad functional repertoire. It hydrolyzes small peptide mediators, modulates cell motility and adhesion to extracellular matrix and also acts as a viral receptor. In order to dissect the function of enzymatically active and inactive APN/CD13, substitutions of different enzymatic active amino acid residues were generated by site-directed mutagenesis and stably transfected into human embryonic kidney cells. All APN variants analyzed exhibited a complete loss of enzymatic activity, whereas wild type APN transfectants exerted a strong aminopeptidase-specific activity. Furthermore, wild type APN expression was associated with a significant decrease in proliferation, migration and also reduced anchorage-independent growth when compared to enzymatically inactive APN variants and controls. This appeared to be due to a downregulated mRNA and protein expression of the chemokine receptor CXCR4 and an inhibition of the stromal cell-derived factor (SDF)-1α/CXCL12-mediated migration. Thus, high APN enzyme activity may antagonize the cellular properties regulated by the CXCR4/SDF-1α system in embryonic kidney cells.     

Biogeography and Host Fidelity of Bacterial Communities in Ircinia spp. from the Bahamas

Research on sponge microbial assemblages has revealed different trends in the geographic variability and specificity of bacterial symbionts. Here, we combined replicated terminal-restriction fragment length polymorphism (T-RFLP) and clone library analyses of 16S rRNA gene sequences to investigate the biogeographic and host-specific structure of bacterial communities in two congeneric and sympatric sponges: Ircinia strobilina, two color morphs of Ircinia felix and ambient seawater. Samples were collected from five islands of the Bahamas separated by 80 to 400 km. T-RFLP profiles revealed significant differences in bacterial community structure among sponge hosts and ambient bacterioplankton. Pairwise statistical comparisons of clone libraries confirmed the specificity of the bacterial assemblages to each host species and differentiated symbiont communities between color morphs of I. felix. Overall, differences in bacterial communities within each host species and morph were unrelated to location. Our results show a high degree of symbiont fidelity to host sponge across a spatial scale of up to 400 km, suggesting that host-specific rather than biogeographic factors play a primary role in structuring and maintaining sponge–bacteria relationships in Ircinia species from the Bahamas.     

Single-cell paired-end genome sequencing reveals structural variation per cell cycle

The nature and pace of genome mutation is largely unknown. Because standard methods sequence DNA from populations of cells, the genetic composition of individual cells is lost, de novo mutations in cells are concealed within the bulk signal and per cell cycle mutation rates and mechanisms remain elusive. Although single-cell genome analyses could resolve these problems, such analyses are error-prone because of whole-genome amplification (WGA) artefacts and are limited in the types of DNA mutation that can be discerned. We developed methods for paired-end sequence analysis of single-cell WGA products that enable (i) detecting multiple classes of DNA mutation, (ii) distinguishing DNA copy number changes from allelic WGA-amplification artefacts by the discovery of matching aberrantly mapping read pairs among the surfeit of paired-end WGA and mapping artefacts and (iii) delineating the break points and architecture of structural variants. By applying the methods, we capture DNA copy number changes acquired over one cell cycle in breast cancer cells and in blastomeres derived from a human zygote after in vitro fertilization. Furthermore, we were able to discover and fine-map a heritable inter-chromosomal rearrangement t(1;16)(p36;p12) by sequencing a single blastomere. The methods will expedite applications in basic genome research and provide a stepping stone to novel approaches for clinical genetic diagnosis.     

The influence of host demography, pathogen virulence, and relationships with pathogen virulence on the evolution of pathogen transmission in a spatial context

A major challenge in evolutionary ecology is to explain extensive natural variation in transmission rates and virulence across pathogens. Host and pathogen ecology is a potentially important source of that variation. Theory of its effects has been developed through the study of non-spatial models, but host population spatial structure has been shown to influence evolutionary outcomes. To date, the effects of basic host and pathogen demography on pathogen evolution have not been thoroughly explored in a spatial context. Here we use simulations to show that space produces novel predictions of the influence of the shape of the pathogen’s transmission–virulence tradeoff, as well as host reproduction and mortality, on the pathogen’s evolutionary stable transmission rate. Importantly, non-spatial models predict that neither the slope of linear transmission–virulence relationships, nor the host reproduction rate will influence pathogen evolution, and that host mortality will only influence it when there is a transmission–virulence tradeoff. We show that this is not the case in a spatial context, and identify the ecological conditions under which spatial effects are most influential. Thus, these results may help explain observed natural variation among pathogens unexplainable by non-spatial models, and provide guidance about when space should be considered. We additionally evaluate the ability of existing analytical approaches to predict the influence of ecology, namely spatial moment equations closed with an improved pair approximation (IPA). The IPA is known to have limited accuracy, but here we show that in the context of pathogens the limitations are substantial: in many cases, IPA incorrectly predicts evolution to pathogen-driven extinction. Despite these limitations, we suggest that the impact of ecology can still be understood within the conceptual framework arising from spatial moment equations, that of “self-shading’’, whereby the spread of highly transmissible pathogens is impeded by local depletion of susceptible hosts.     

Parkin-Dependent Degradation of the F-Box Protein Fbw7β Promotes Neuronal Survival in Response to Oxidative Stress by Stabilizing Mcl-1

Parkinson''s disease (PD) is characterized by progressive loss of midbrain dopaminergic neurons resulting in motor dysfunction. While most PD is sporadic in nature, a significant subset can be linked to either dominant or recessive germ line mutations. PARK2, encoding the ubiquitin ligase parkin, is the most frequently mutated gene in hereditary Parkinson''s disease. Here, we present evidence for a neuronal ubiquitin ligase cascade involving parkin and the multisubunit ubiquitin ligase SCF^Fbw7β. Specifically, parkin targets the SCF substrate adapter Fbw7β for proteasomal degradation. Furthermore, we show that the physiological role of parkin-mediated regulation of Fbw7β levels is the stabilization of the mitochondrial prosurvival factor Mcl-1, an SCF^Fbw7β target in neurons. We show that neurons depleted of parkin become acutely sensitive to oxidative stress due to an inability to maintain adequate levels of Mcl-1. Therefore, loss of parkin function through biallelic mutation of PARK2 may lead to death of dopaminergic neurons through unregulated SCF^Fbw7β-mediated ubiquitylation-dependent proteolysis of Mcl-1.     

Using plant functional traits to explain community composition across a strong environmental filter in Australian alpine snowpatches

Environmental filters act to limit the local community assemblage from the regional species pool by restricting the viable trait states that can occur there. In alpine snowpatches, the timing of snowmelt is a strong environmental filter. In coming decades, the strength of this filter is likely to relax with global climate change. We used three continuous plant functional traits (leaf area, plant height, seed mass) and their divergence (using the FD_var index) to document current patterns of community assembly and predict plant community responses to future environmental filters in alpine snowpatch vegetation. The community trait-weighted mean for leaf area and height, but not seed mass, was significantly higher in early snowmelt zones relative to mid and late melting zones across all snowpatches. Mean FD_var for height (but not leaf area or seed mass), by contrast, was substantially lower in early snowmelt zones, indicating that species growing in early melt zones are consistently taller than those growing in other zones. These results suggest that if climate change leads to earlier snowmelt and hence, a longer growing season, taller (more competitive) species with larger leaf areas (more productive) may replace short species in snowpatches as these plant communities re-assemble in response to changing environmental filters.     

Pyrrolo[1,2-a]pyrazine and pyrazolo[1,5-a]pyrazine: Novel,potent, and selective series of Vasopressin1b receptor antagonists

Novel series of pyrrole-pyrazinone and pyrazole-pyrazinone have been identified as potent and selective Vasopressin_1b receptor antagonists. Exploration of the substitution pattern around the core of these templates allowed generation of compounds with high inhibitory potency at the Vasopressin_1b receptor, including examples that showed good selectivity with respect to Vasopressin_1a, Vasopressin₂, and Oxytocin receptor subtypes.