Evolution and maintenance of haploid–diploid life cycles in natural populations: The case of the marine brown alga Ectocarpus

The evolutionary stability of haploid–diploid life cycles is still controversial. Mathematical models indicate that niche differences between ploidy phases may be a necessary condition for the evolution and maintenance of these life cycles. Nevertheless, experimental support for this prediction remains elusive. In the present work, we explored this hypothesis in natural populations of the brown alga Ectocarpus. Consistent with the life cycle described in culture, Ectocarpus crouaniorum in NW France and E. siliculosus in SW Italy exhibited an alternation between haploid gametophytes and diploid sporophytes. Our field data invalidated, however, the long‐standing view of an isomorphic alternation of generations. Gametophytes and sporophytes displayed marked differences in size and, conforming to theoretical predictions, occupied different spatiotemporal niches. Gametophytes were found almost exclusively on the alga Scytosiphon lomentaria during spring whereas sporophytes were present year‐round on abiotic substrata. Paradoxically, E. siliculosus in NW France exhibited similar habitat usage despite the absence of alternation of ploidy phases. Diploid sporophytes grew both epilithically and epiphytically, and this mainly asexual population gained the same ecological advantage postulated for haploid–diploid populations. Consequently, an ecological interpretation of the niche differences between haploid and diploid individuals does not seem to satisfactorily explain the evolution of the Ectocarpus life cycle.     

Dubinectes infirmus, a new species of deep-water Munnopsidae (Crustacea, Isopoda, Asellota) from the Argentine Basin, South Atlantic Ocean

Dubinectes infirmussp. n., Munnopsidae, is described from the Argentine Basin, southwest Atlantic, at depths between 4586-4607 m. The new species is distinguished by a narrow rim of the pleotelson posterior margin which is not raising over its dorsal surface; article 3 of the antennula is subequal in length to article 2; distomedial lobes of male pleopod 1 are of same size as distolateral lobes; stylet of male pleopod 2 is subequal in length to protopod; uropod exopod is more than a half of endopod length. Some generic characters which are weakly pronounced in the new species or have different state are defined more precisely in the revised diagnosis of Dubinectes. The modified diagnosis of the genus, a key to the species of Dubinectes as well as the distribution of the genus are presented.     

Militarised natural history: tales of the avocet's return to postwar Britain

Absent as a breeding bird from Britain for at least a century, avocets (Recurvirostra avosetta) began nesting on the east coast of Britain, in Suffolk, shortly after the end of the Second World War, having honed in on two spots on Britain's coast that had been flooded for war-related reasons. The avocets' presence was surrounded in secrecy, while a dedicated few kept up a protective watch over them. As the Royal Society for the Protection of Birds (RSPB) took over responsibility for the flourishing colony, they claimed the episode as a symbol of success for British protection, later making the bird their logo. Counter to the RSPB's story of protecting a British bird, I read the narratives of events in terms of making a bird British. I show how, as postwar Britain slumped economically and spiritually and tried to rebuild itself, the birds became a vehicle for formulating national identity: of Britain as a home to which to return and belong. Exploring the themes of returning servicemen and closed territories, the paper also examines the episode in terms of the naturalisation of the military and the militarisation of nature.     

Surrogate antigens as targets for proteome-wide binder selection

In the past decade, many initiatives were taken for the development of antibodies for proteome-wide studies, as well as characterisation and validation of clinically relevant disease biomarkers. Phage display offers many advantages compared to antibody generation by immunisation because it is an unlimited resource of affinity reagents without batch-to-batch variation and is also amendable for high throughput in contrast to conventional hybridoma technology. One of the major bottlenecks to proteome-wide binder selection is the limited supply of suitable target antigens representative of the human proteome. Here, we provide proof of principle of using easily accessible, cancer-associated protein epitope signature tags (PrESTs), routinely generated within the Human Protein Atlas project, as surrogate antigens for full-length proteins in phage selections for the retrieval of target-specific binders. These binders were subsequently tested in western blot, immunohistochemistry and protein microarray application to demonstrate their functionality.     

Translational database selection and multiplexed sequence capture for up front filtering of reliable breast cancer biomarker candidates

Biomarker identification is of utmost importance for the development of novel diagnostics and therapeutics. Here we make use of a translational database selection strategy, utilizing data from the Human Protein Atlas (HPA) on differentially expressed protein patterns in healthy and breast cancer tissues as a means to filter out potential biomarkers for underlying genetic causatives of the disease. DNA was isolated from ten breast cancer biopsies, and the protein coding and flanking non-coding genomic regions corresponding to the selected proteins were extracted in a multiplexed format from the samples using a single DNA sequence capture array. Deep sequencing revealed an even enrichment of the multiplexed samples and a great variation of genetic alterations in the tumors of the sampled individuals. Benefiting from the upstream filtering method, the final set of biomarker candidates could be completely verified through bidirectional Sanger sequencing, revealing a 40 percent false positive rate despite high read coverage. Of the variants encountered in translated regions, nine novel non-synonymous variations were identified and verified, two of which were present in more than one of the ten tumor samples.     

Engineering bispecificity into a single albumin-binding domain

Bispecific antibodies as well as non-immunoglobulin based bispecific affinity proteins are considered to have a very high potential in future biotherapeutic applications. In this study, we report on a novel approach for generation of extremely small bispecific proteins comprised of only a single structural domain. Binding to tumor necrosis factor-α (TNF-α) was engineered into an albumin-binding domain while still retaining the original affinity for albumin, resulting in a bispecific protein composed of merely 46 amino acids. By diversification of the non albumin-binding side of the three-helix bundle domain, followed by display of the resulting library on phage particles, bispecific single-domain proteins were isolated using selections with TNF-α as target. Moreover, based on the obtained sequences from the phage selection, a second-generation library was designed in order to further increase the affinity of the bispecific candidates. Staphylococcal surface display was employed for the affinity maturation, enabling efficient isolation of improved binders as well as multiparameter-based sortings with both TNF-α and albumin as targets in the same selection cycle. Isolated variants were sequenced and the binding to albumin and TNF-α was analyzed. This analysis revealed an affinity for TNF-α below 5 nM for the strongest binders. From the multiparameter sorting that simultaneously targeted TNF-α and albumin, several bispecific candidates were isolated with high affinity to both antigens, suggesting that cell display in combination with fluorescence activated cell sorting is a suitable technology for engineering of bispecificity. To our knowledge, the new binders represent the smallest engineered bispecific proteins reported so far. Possibilities and challenges as well as potential future applications of this novel strategy are discussed.     

Comparative analysis of algal biodiversity in the rivers of Israel

Comparative analysis of algal communities in the rivers of Israel was completed to highlight the influence of environmental variables on biodiversity. The study revealed that 671 species of algae and cyanobacteria belonging to nine taxonomic divisions were present during 2002–2009 in the Yarqon, Alexander, Hadera, Qishon, Oren, Lower and Upper Jordan, and Zin rivers. The species richness of each river was evaluated by taxonomic structural comparison, geobotanical, hierarchical cluster analysis, and the degree of relatedness for different levels of taxonomic resolution. The analysis revealed close similarity of the Upper Jordan and Oren rivers. The average taxonomic distinctness index showed that the Yarqon, Oren, Upper Jordan, and Qishon communities were partly degraded due to permanent environmental disturbances. The variation in taxonomic distinctness index showed that the Alexander, Yarqon and Hadera communities were formed not only due to anthropogenic factors but also through long-term climatic impact. The most abundant indicator species inhabit low streaming and standing alkaline waters of medium salinity and low to medium organic pollution. The statistical approaches allowed discrimination between climatic and anthropogenic factors that impact upon the riverine biodiversity in semi-arid environments. Analysis shows the influence of anthropogenic factors was strongly modulated by climatic impacts causing a marked decease of species richness from north to south.     

Reversible resistance induced by FLT3 inhibition: a novel resistance mechanism in mutant FLT3-expressing cells

Objectives

Clinical responses achieved with FLT3 kinase inhibitors in acute myeloid leukemia (AML) are typically transient and partial. Thus, there is a need for identification of molecular mechanisms of clinical resistance to these drugs. In response, we characterized MOLM13 AML cell lines made resistant to two structurally-independent FLT3 inhibitors.

Methods

MOLM13 cells were made drug resistant via prolonged exposure to midostaurin and HG-7-85-01, respectively. Cell proliferation was determined by Trypan blue exclusion. Protein expression was assessed by immunoblotting, immunoprecipitation, and flow cytometry. Cycloheximide was used to determine protein half-life. RT-PCR was performed to determine FLT3 mRNA levels, and FISH analysis was performed to determine FLT3 gene expression.

Results and Conclusions

We found that MOLM13 cells readily developed cross-resistance when exposed to either midostaurin or HG-7-85-01. Resistance in both lines was associated with dramatically elevated levels of cell surface FLT3 and elevated levels of phosphor-MAPK, but not phospho-STAT5. The increase in FLT3-ITD expression was at least in part due to reduced turnover of the receptor, with prolonged half-life. Importantly, the drug-resistant phenotype could be rapidly reversed upon withdrawal of either inhibitor. Consistent with this phenotype, no significant evidence of FLT3 gene amplification, kinase domain mutations, or elevated levels of mRNA was observed, suggesting that protein turnover may be part of an auto-regulatory pathway initiated by FLT3 kinase activity. Interestingly, FLT3 inhibitor resistance also correlated with resistance to cytosine arabinoside. Over-expression of FLT3 protein in response to kinase inhibitors may be part of a novel mechanism that could contribute to clinical resistance.